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Bekisz M., Sławińska U., Kwaśniewska A., Miazga K., Zawadzka M.
Laboratory of Neuromuscular Plasticity, Nencki Institute of Experimental Biology, Polish Academy of Science, Warsaw, Poland
Abstract: Different environmental conditions activate diverse differentiation pathways in progenitor cells thus different types of cells could be generated. To understand the influence of specific environmental conditions on embryonic progenitor cells (EPCs) fate, the cells dissected from the embryonic brainstem were grown in vitro in the presence of either Shh/FGF4 (S) or EGF/bFGF (E). Isolated cells effectively proliferated, creating neurospheres (S1 and E1), capable to form secondary neurospheres (S2) or monolayers (E2). Bioinformatic analysis of transcriptomes showed significant differences in gene expression of EPCs grown under different conditions. We found that in S1 the most upregulated genes were related to neurogenesis and nervous system development as well as axon growth processes, while in S2 the upregulated genes were related to the organization of genetic material in the cell as chromatin and nucleosome formation. Higher expression of genes involved in neurogenesis was also found in the E1 vs E2 cells. Next, the differentiation fate of pre-treated groups of cells either in neural or glia-promoting medium was investigated, which also indicated significant differences in their progeny characteristics.
Concluding, EPCs adopt different phenotypes in vitro. Whether neural progenitor cells would be able to differentiate in specific in vivo conditions should be investigated next.
Funding:Supported by the Nencki Institute statutory donation and by ERA-NET NEURON CoFund Consortium NEURONICHE (The National Centre for Research and Development; ERA-NET-Neuron/16/17).
Weronika Maksymiuk1, Justyna Gargas1, Monika Sypecka2, Anna Sarnowska2, Joanna Sypecka1
1NeuroRepair Department, Mossakowski Medical Research Institute Polish Academy of Sciences, Warsaw, Poland
2Translational Platform for Regenerative Medicine, Mossakowski Medical Research Institute Polish Academy of Sciences, Warsaw, Poland
Abstract: Brain damage caused by neuronal and glial cell death or dysfunction leads to several neurodevelopmental disorders, such as aberrant formation of the brain's white matter. The search for an effective therapy began with attempts to differentiate human mesenchymal stem/stromal cells (MSCs) into glial lineage cells. MSCs, obtained from the Wharton's Jelly (WJ) of the umbilical cord up to 24 hours after birth, were at first cultured under physiologically normoxic conditions, in standard culture medium. On the second day after seeding, the medium was changed to induce glial commitment of the cultured cells. Accordingly, MSCs were cultured at different culture densities with the addition of the mitogen PDGF-AA to the culture medium or in the serum-free medium conditioned by mixed primary glial culture derived from neonatal rats. After 2-3 weeks in differentiating medium, the cells were used for morphological, immunocytochemical and molecular analyses to verify their phenotype. The preliminary results indicate that WJ-MSCs have the potential to undergo the differentiation process and will be used to elaborate a protocol for the efficient derivation of cells of glial phenotype from WJ-MSCs.
Funding: Supported by National Science Center (NCN) in Poland, grant: 2022/47/O/NZ4/01161.
Miazga K., Sławińska U., Bekisz M., Zawadzka M.
Laboratory of Neuromuscular Plasticity, Nencki Institute of Experimental Biology, Polish Academy of Science, Warsaw, Poland
Abstract: In the adult mammalian spinal cord, the central canal ependymal zone is a niche of neural progenitor cells (ependymal NPCs) that might be a potential source of cells useful in therapy of spinal cord injuries. In lower vertebrates after spinal cord injury, those cells proliferate and differentiate into neurons which participate in the restitution of lost motor function. However, in mammals after spinal cord injury ependymal cells differentiate mainly into astrocytes forming a glial scar that often limits neural regeneration.
The aim of our study was to identify genes and/or signaling pathways that can potentially control the proliferation and differentiation of rat spinal cord ependymal NPCs after injury.
To achieve that we selectively collected ependymal cells from the tissue sections of the spinal cord central canal of injured and control rats and compared their transcriptomes. Bioinformatic analysis identified genes specifically upregulated in ependymal cells that participate in the control of TGF-β and WNT signaling pathways in injured rat spinal cords. Moreover, we identified Klf5 and Elf3 as two transcription factors uniquely upregulated in central canal cells. Understanding the molecular mechanisms involved in the proliferation and differentiation of ependymal NPCs may contribute to the development of new therapies for spinal cord injury treatment based on the manipulation of their fate.
Funding: National Science Center MINIATURA 4 Grant(2020/04/X/NZ3/00185)
P. Gebala, J. Janowska, H. Zajac, J. Sypecka
NeuroRepair Department, Mossakowski Medical Research Institute, Polish Academy of Sciences, Warsaw, Poland
Abstract: The process of autophagy is involved in the survival and maintenance of every cell. Its disruption, especially deficits, can lead to pathophysiological changes in tissues and cause various diseases, including cancer and neurodegenerative disorders. Recently, it has been shown that dysregulation of autophagy may be involved in the brain damage and hypomyelination that develop after neonatal asphyxia.
In this study, we investigated the process of autophagy in primary cultures of neonatal rat oligodendrocytes and oligodendrocyte progenitor cells (OPCs). We exposed them to different oxygen-glucose deprivation (OGD) procedures to mimic hypoxia-ischemia in the neonatal brain. To inhibit lysosomal degradation, we tested chloroquine treatment of cell cultures.
OGD performed on OPCs and lasting 40 minutes did not induce massive autophagy when p62 and LC3-II were detected by immunoblot. The 24 h incubation with 25 - 100 uM chloroquine induced cell death, while 6 h incubation at 5 - 25 uM concentrations efficiently inhibited autophagic flux and affected oligodendrocyte differentiation.
Further research will shed light on the prospect of using pharmaceuticals that modulate the autophagy process to treat the consequences of neonatal asphyxia.
Funding: Supported by National Science Center in Poland, grant: 2021/03/Y/NZ4/00214.
Aleksandra Rzeszut1, Julia Netczuk1, Zofia Harda1, Klaudia Misiołek1, Łukasz Szumiec1, Barbara Ziółkowska1, Jan Rodriguez Parkitna1
1Department of Molecular Neuropharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences, Kraków, Poland
Abstract: Maintaining complex social relationships is a fundamental ability for humans and other group living animals. Social dysfunction is present in many neuropsychiatric disorders. Social memory, i.e. the ability to recognize familiar conspecifics, plays a crucial role in maintaining appropriate social relationships.
Previous studies indicate the involvement of the kappa opioid system in the formation of social memory traces. In particular, it was demonstrated that prodynorphin, κ opioid receptor (KOR) ligand, knockout mice (Pdyn -/-) retain social memory traces longer than their wild-type littermates. However, the mechanism underlying this phenomenon is not known. The question remains which of the KOR expressing neurons (serotonergic or oxytocinergic) react to dynorphin in a way that erases the social memory trace. To test this, we examined social memory in two mouse strains with selective inactivation of KOR receptors on serotonin or oxytocin expressing neurons (Oprk1Tph2CreERT2 and Oprk1OxtCre strain, respectively). In our preliminary experiments Oprk1OxtCre mice, but not Oprk1Tph2CreERT2, maintained a social memory trace for a longer period than animals from the control group, recapitulating the Pdyn -/- mice phenotype. These results suggest that the modulation of oxytocin signaling by the dynorphin/KOR system plays a major role in the erasure of social memory trace. Therefore, further research is necessary to fully understand the role of the interplay between dynorphin and oxytocin signaling in social memory retention.
Funding: National Science Centre, Poland OPUS 2019/35/B/NZ7/03477.
Karolina Protokowicz, Leszek Kaczmarek
Laboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology, PAS, Warsaw Poland
Abstract: Our study investigates the role of matrix metalloproteinase-9 (MMP-9) an extracellular protease, a key player in synaptic plasticity, in behavioral consequences of early postnatal inflammation.
We administered a single injection of bacterial lipopolysaccharide (LPS, 0.05 mg/kg) or saline to postnatal-day-7 mice. Two hours post-administration, TIMP-1 serum levels were elevated in both sexes compared to saline controls. MMP-9 levels were elevated in males but not females treated with LPS, as demonstrated by Luminex® immunoassay. Brain MMP-9 activity was assessed using gel zymography protocol, revealing heightened activity in the hippocampi of both sexes two hours post-LPS injection. After six hours, elevated MMP-9 activity was observed only in male cortices. To further investigate MMP-9's role, we conducted behavioral assessments on adult wild-type (WT) animals and MMP-9 knockout (KO) littermates following LPS injection on P7. WT males exhibited a decreased interest in odors from unfamiliar animals but increased socializing with familiar cage mates post-immune challenge. In contrast, WT females displayed heightened interest in unfamiliar social stimuli and reduced sociability within known groups. Remarkably, these effects were absent in MMP-9 KO animals. Our results suggest MMP-9's involvement in long-term sociability alterations following immune activation, but further investigation into underlying molecular mechanisms is necessary.
Funding: BRAINCITY - Centre of Excellence for Neural Plasticity and Brain Disorders’ project of Foundation for Polish Science (FNP).
Anna Janus1, Klaudia Lustyk1, Agata Siwek2, Henryk Marona3, Karolina Pytka1
1Department of Pharmacodynamics, Jagiellonian University Medical College, Cracow, Poland
2Department of Pharmacobiology Jagiellonian University Medical College, Cracow, Poland
3Department of Bioorganic Chemistry, Chair of Organic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Cracow, Poland
Abstract: Serotonin 5-HT1A receptors have recently gained a lot of attention in the field of neuropharmacology due to their association with the neuropathology of depression, anxiety, and cognitive impairments. Several studies suggest that xanthone derivatives ofpiperazine, which possess an affinity for 5-HT1A receptors, exhibit antidepressant-like effects. Considering that despite years of investigation, research on antidepressant drugs still poses a significant challenge, here we aimed to evaluate the preliminary pharmacological properties of HBK-1a, a novel xanthone derivative of piperazine. In the in vitro radioligand binding assays, we assessed the affinity of HBK-1a for 5-HT1A receptors in the rat hippocampus and for adrenergic α1 and β1 receptors in the rat cortex. Moreover, we investigated its ability to induce antidepressant-like effects in the forced swim test and its influence on spontaneous locomotor activity in mice. Our results demonstrate that HBK-1a shows moderate affinity towards 5-HT1A and α1-adrenergic receptors. At a dose of 10mg/kg, HBK-1a significantly reduced the immobility time in the forced swim test. The compound did not influence spontaneous locomotor activity in mice, which excludes the possibility of changes in locomotor activity influencing the results. Our findings confirm that piperazine xanthone derivatives exhibit antidepressant-like properties, however further research is required.
Kalinowska M.1, Yadav A.1, Haque F.1,2, Bryksa A.1, Chen X.3, Rycerz M.1, Sadowska J.1, Puścian A.1
1Laboratory of Emotions Neurobiology, BRAINCITY- Centre of Excellence for Neural Plasticity and Brain Disorders, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
2Molecular Imaging Branch- National Cancer Institute, Bethesda, MD, USA
3Laboratoire de Physique de l'EcoleNormaleSuperieure, The French National Centre for ScientificResearch
Abstract: Social mammals - including people – function as a part of a group for most of their lives. However, social relationships change over time. The ability to assess the dynamics of such shifts is crucial for understanding of neuronal mechanisms underlying social relationships.
To investigate the stability of social bonds over time, we used groups of mice kept in the computer-controlled experimental environment (Eco-HAB) in which they could move freely form social bonds. Eco-HAB system enables measuring the time each pair of mice within the group spends together voluntarily on each day, as reflected by the variable called in-cohort sociability. Further, we used this measure to design the autocorrelation-based algorithm for the assessment of the stability of the social bond between any given two animals over time.
We show that mouse dyads do form stable relationships. Moreover, about 2/3 of the tested pairs developed constant level of spending time together. Additionally, we show that for all individual animals, the mean values of in-cohort sociability are very close to each other, which means that all animals form social bonds to the same extent, however, with different individuals.
We present an approach to measuring the formation of social bonds in animals over time.
Wróblewski K.1,5, Bryksa A.1, Haque F.1, Kondrakiewicz L.1,2, Winiarski M.1, Kasprowicz G.3, Rupp-Janecka D.1, Jędrzejewska-Szmek J.4, Turzyński K.4, Knapska E.1, Puścian A.1,
1Laboratory of Emotions Neurobiology, BRAINCITY - Centre of Excellence for Neural Plasticity and Brain Disorders, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
2Haesler Lab, NeuroElectronics Research Flanders, KU Leuven
3Warsaw University of Technology, Warsaw, Poland
4Laboratory of Neuroinformatics, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
5Faculty of Physics, University of Warsaw, Warsaw, Poland
Abstract: To accelerate the research on spontaneous social behaviors that reliably activate specific, evolutionarily established neural pathways, we developed a new field assay and accompanying analytical tools.
Eco-HAB Field is an automated-testing environment located in the wild habitat. It consists of 8 subterritories, connected via underground corridors equipped with the RFID antennas, providing continuous recording of behavior in groups of mice. It enables testing behavioral patterns relevant for surviving and thriving in varying natural conditions, as well as social interactions and their stability over time with high ecological accuracy.
We show that the developed methodology not only provides access to naturalistic patterns of behavior under naturalistic conditions but also is characterized by high data collection validity. To demonstrate the latter, we developed custom Python scripts for analyzing the potential antennas' misreadings and skipped registrations, thus determining influence of such events on reliability of the collected data. We established that corrupted data segments are rare, relatively evenly distributed over time, and constitute less than 1% of total registrations.
Our results confirm Eco-HAB Field as a reliable approach to studying neural underpinnings of behaviors crucial for survival and success across different environmental conditions tested in groups of mice.
A. Bryksa1, F. Haque2, J. Borowska1, K. Nazaruk1, J. Jędrzejewska-Szmek3, S. Łęski3, M. Winiarski1, E. Knapska1, A. Puścian1
1Laboratory of Emotions Neurobiology, BRAINCITY - Centre of Excellence for Neural Plasticity and Brain Disorders, Nencki Institute, Polish Academy of Sciences, Warsaw, Poland
2Artificial Intelligence Resource, Molecular Imaging Branch, National Cancer Institute, Bethesda, MD, USA
3Laboratory of Neuroinformatics, Nencki Institute of Experimental Biology of Polish Academy of Sciences, Warsaw, Poland
Abstract: Understanding the role of the prelimbic cortex (PL) in encoding social attachment remains a challenge. This study investigates the functional role of various neuronal classes in the PL, including pyramidal cells (Pyr), parvalbumin- (PV+), somatostatin- (Sst+), and vasoactive intestinal peptide- (VIP+) expressing interneurons, in the processing of familiar and novel social stimuli.
We employed chemo/optogenetics and automated behavioral testing in Eco-HAB® system, which allows automated, individualized measurement of voluntary behavior, while manipulating the activity of specific neuronal populations in the PL. Our evaluation focused on animals' responses to known conspecifics and their interest in novel social stimuli.
The results indicate that inhibiting Pyr and activating PV+ interneurons in the PL reduce sociability by diminishing the time voluntarily spent with cagemates. Interestingly, Pyr inhibition also increases interest in novel social stimuli, a behavior not observed in animals with activated PV+. Furthermore, VIP+ cell activation mirrors the functional role of the PV+ but to a lesser extent. Conversely, Sst+ activation disrupts interest in both familiar and novel social stimuli indiscriminately.
These findings underscore the diverse roles of different PL neuronal classes in processing familiar and novel social information. Importantly, the study emphasizes the crucial role of PV+ neurons in selectively encoding familiarity.
Funding: Nencki Institute, BRAINCITY – Center of Excellence for Neural Plasticity and Brain Disorders, NCN 2020/39/D/NZ4/01785, Kosciuszko Foundation
Bettina U. Wilke¹, Maryke Benz², Sonja Diehl¹, Eliza Koros¹, Wiebke Nissen¹, Susanne Zach¹, Roberto Arban¹, Christopher Pryce³ and Judith Schweimer¹
¹Boehringer Ingelheim Pharma GmbH & Co. KG, CNS DR, Biberach, Germany
²Boehringer Ingelheim Pharma GmbH & Co. KG, DDS, Biberach, Germany
³Department for Psychiatry, Psychotherapy and Psychosomatics, University of Zurich, Zürich, Switzerland
Abstract: Altered central serotonin (5-HT) signalling has been associated with anxiety disorders, among other clinical indications. Human studies indicate a positive correlation between severity of social anxiety symptoms and 5-HT synthesis in the amygdala and dorsal anterior cingulate cortex (ACC). Polymorphisms in the gene encoding tryptophan hydroxylase (Tph2), the rate-limiting enzyme for 5-HT synthesis, are not only linked to elevated 5-HT levels in the amygdala and ACC, but also associated with increased amygdala reactivity to emotional stimuli. Animal studies further support the idea that alterations in 5-HT neurotransmission cause changes in threat responsiveness.
Tph2 gene expression in the dorsal raphe nucleus (DRN) is reported to be upregulated by acute stress. Whereas chronic social stress increases Htr1a and Slc6a4, the gene that encodes the 5-HT transporter (SERT), expression in the DRN, amongst other 5-HT changes. In this study, we aimed to investigate in mice if short-term exposure to social aversion that results in adaptive social avoidance would also lead to elevated tissue levels of Tph2 mRNA in the midbrain raphe nucleus and of levels of 5-HT and its major metabolite in projection regions involved in emotional stimulus processing.
Bartosz Zglinicki, Patrycja Ziuzia andMichał Ślęzak
Biology of Astrocytes Group, Life Sciences & Biotechnology Center, Lukasiewicz Research Network – PORT Polish Center for Technology Development, Wroclaw, Poland
Abstract: Validation of any biological intervention developed for tackling the root or symptoms of neuropsychiatric disorders requires proper tools. Disturbances of social behavior is a hallmark of many psychiatric conditions, such as depression, anxiety and schizophrenia. Capturing and quantifying broad range of social behavior simultaneously within a single tool is therefore essential, as it would allow for registration and clustering of groups of behavior specific for particular disorders. Such method would be of great advantage for better translational studies.
Here, within Same-NeuroID project, we implemented a pipeline for efficient characterization of complex social behavior in mice. We call it Same-SocialBox. In the setup animals are housed together in a semi-naturalistic environment with proper bedding, food and water access, and with night-day cycle. Animals are recorded for long hours to ensure a capture of diverse behavior. SLEAP.ai is used for pose estimation of recorded animals and extracted coordinates are processed by deepOF software to create set of features. These features are then used to “feed” models of both supervised and unsupervised learning for complex behavior classification.
Funding: This work was supported by the NCN OPUS grant 2021/41/B/NZ3/04099 'AstroSyCo' and HE Twinning 'SAME-NeuroID' grant No: 101079181.
Aleksandra Herud1, Sedef Dalbeyler2, Laura Bergauer2, Marcin Piechota3,4, Michał Korostyński3,4, Michał Ślęzak1,2
1Biology of Astrocytes Research Group, Łukasiewicz Research Network – PORT Polish Institute for Technology Development, Wrocław, Poland
2BioMed X Institute, Heidelberg, Germany
3Laboratory of Pharmacogenomics, Department of Molecular Neuropharmacology, Maj Institute of Pharmacology, PAS, Kraków, Poland
4Intelliseq, Kraków, Poland
Abstract: Stress is an adaptive reaction of the organism to alterations of the environment. The stress response is coordinated by the hypothalamus-pituitary-adrenal (HPA) axis, largely through secretion of glucocorticoids (GCs). Glucocorticoids receptors (GRs) act primarily as transcription factors activated by increased levels of GCs, such as those observed upon stress. Growing evidence suggest that astrocytes play an important role in mediating central effects of GCs. Astrocytes sense local synaptic activity, they regulate neurotransmitter homeostasis, and signal back to the synapse, securing physiological performance of neural networks. Our group showed that GR deletion in astrocytes affects the formation of aversive memory, which correlated with the changes in the expression of metabolic genes. Here, we examined GR-dependent transcriptional effects of chronic social defeat stress in astrocytes. We isolated astrocytes from 2 brain regions relevant for stress response and found differential effects of CSDS in the two brain structures examined, with certain overlap. We identified a significant contribution of the GR to these changes. Furthermore, elimination of GR from astrocytes abolished vast majority of CSDS-induced changes, while other molecular pathways, previously unaltered by CSDS, were affected in GR astroKO mice. We conclude that GR signaling in astrocytes mediates transcriptional effects of chronic stress in these cells.
Funding: This work was supported by the NCN OPUS grant 2021/41/B/NZ3/04099 'AstroSyCo'.
Aniqa Saiyara1, Aleksandra Koszałka1, Kinga Sałaciak1, Karolina Pytka1
1Department of Pharmacodynamics, Faculty of Pharmacy, Jagiellonian University Medical College, Krakow, Poland
Abstract: Depression, a highly prevalent mental health condition, presents a significant challenge to healthcare, especially for women, who experience twice the prevalence compared to men. However, despite the availability of various treatment options, there are notable variations in treatment responses.
In this study, using the unpredictable chronic mild stress (UCMS) model, we aimed to assess the efficacy of vortioxetine in female mice, focusing on its effects on recognition memory and anhedonic behavior.
Female C57BL/6 mice underwent 6 weeks of UCMS, while a control group remained unstressed. Daily administration of vortioxetine at a dosage of 5 mg/kg began during the final 14 days of the study. Depressive-like behavior was assessed using the sucrose preference test, while cognition was evaluated using the object recognition test.
We observed the successful induction of anhedonia after the third week of UCMS, with a more pronounced effect evident by the fourth week. Furthermore, repeated administration of vortioxetine effectively reversed anhedonia in female mice. However, an antidepressant did not attenuate recognition memory deficits.
While our findings indicate the potential of vortioxetine to reverse anhedonia in female mice, its effect on memory was not present, suggesting possible differences in the treatment response depending on the sex.
Funding:This study was conducted as part of a research project funded by the National Science Centre, Poland (grant 2019/34/E/NZ7/00454).
Klaudia Misiołek1, Zofia Harda1, Magdalena Chrószcz1, Marta Klimczak1, Łukasz Szumiec1, Aleksandra Rzeszut1, Barbara Ziółkowska1, Jan Rodriguez Parkitna1
1Maj Institute of Pharmacology Polish Academy of Sciences, Krakow, Poland
Abstract: Adolescence is a period of intense brain reorganization, accompanied by heightened reward sensitivity. Though endogenous opioid system is implicated in this phenomenon, the specific mechanisms remain elusive. We have discovered that in the social conditioned place preference test (social CPP) the social reward temporarily decrease in mid-adolescence male C57BL/6 mice. This decrease appears to be specific to social domain since we did not observe similar results in another natural reward (food CPP). The social reward result is in contrast to cocaine reward (cocaine CPP) that peaks during mid-adolescence. Treatment with a selective κ-opioid receptor antagonist, norbinaltorphimine (i.p., 10mg/kg), induced a drop in social reward in early adolescent mice. Furthermore, using qPCR we investigated changes in the relative mRNA expression of the μ, δ and κ-opioid receptors (Oprm1, Oprd1 and Oprk1, respectively) and the opioid peptide precursors prodynorphin (Pdyn) and preproenkephalin (Penk) in key areas of the brain’s reward system areas (prefrontal cortex (PFC), nucleus accumbens (NAc), dorsal striatum (DS)). We found a decrease in expression of (mRNA) Pdyn in mid-adolescent mice in the PFC and DS. The relative expression of (mRNA) Oprd1 in NAc appears to increase with age. Taken together, these data shows the developmental changes of sensitivity to the rewards across mice adolescence and point at κ-opioid signalling as potential mechanism shaping adolescent social behaviour.
Funding:This research was funded by the National Science Centre, Poland OPUS 2019/35/B/NZ7/03477 and PRELUDIUM-21 2022/45/N/NZ4/01504.
Sylwia Drabik1, Aleksandra Trenk1, Anna Gugula1, Patryk Sambak1, Angelika Kaleta1, Gabriela Stopka1, Mohammed Akhter Hossain2, Andrew L. Gundlach2, Anna Blasiak1
1Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland
2The Florey Institute of Neuroscience and Mental Health, and Florey Department of Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia
Abstract: The midbrain interpeduncular nucleus (IPN) plays a significant role in social behaviour and anxiety control. IPN neurons highly express the nerve-growth factor (NGF) receptors – TrkA and are densely innervated by the stress-sensitive nucleus incertus (NI). NI is a the primary source of relaxin-3 (RLN3), and it innervates the ventral hippocampus (vHPC), vital for social behaviour and anxiety signalling. However, the relationships within of the NI-IPN-vHPC axis remain unknown.
Fluorescent in situ hybridisation showed that GABAergic IPN neurons co-express TrkA and RLN3 receptor (RXFP3) mRNA. Neural tract-tracing unveiled extensive RLN3 projections from the NI to the IPN. Whole-cell patch clamp recordings demonstrated that RXFP3-selective agonist A2 induces outward currents in IPN neurons, and multielectrode array recordings revealed both excitatory and inhibitory NGF action within the IPN. Furthermore, tract-tracing studies revealed dense IPN originating innervation of the vHPC, particularly from the rostral and lateral nuclei, that at the same time are strongly innervated by the NI.
Taken together, our findings show that NGF-sensitive IPN neurons innervating vHPC receive innervation from NI and remain under the control of RLN3/RXFP3 system, implicating important role of NI-IPN-vHPC axis in the control of social interactions and anxiety.
Funding: National Science Centre, Poland (UMO-2018/30/E/NZ4/00687; UMO 2021/41/N/NZ4/04499; UMO-2023/49/B/NZ4/01885).
Olga Mierniczek1, Gniewosz Drwiega1, Martyna Gorkowska1, Joanna Roszkowska1, Gabriela Izowit1, Wojciech Solecki2, Tomasz Blasiak1
1Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University, Cracow, Poland
2Department of Neurobiology and Neuropsychology, Institute of Applied Psychology, Jagiellonian University, Cracow, Poland
Abstract: The brainstem nucleus incertus (NI) is involved in modulating stress response and aversive stimuli processing. It is also a primary source of the stress related neuropeptide relaxin-3 (RLN-3). NI innervates ventral tegmental area (VTA), a key component of the brain's reward system. By tracing neural pathways using a retrograde viral vector, the NI-originating innervation of the VTA was confirmed within the midbrain of Sprague-Dawley rats. Immunohistochemical staining indicated that only a small percentage of neurons in the NI, which contain the RLN-3, also extend projections to the VTA, implying a specialized connectivity pattern between these regions. Behavioral experiments have shown increased activity in NI neurons, as indicated by c-Fos expression, in response to painful stimuli. This highlights the NI's involvement in processing adverse conditions, bolstering its role in stress response. Electrophysiological recordings further reveal excitatory reactions of NI neurons to painful stimuli, and intriguingly, the response direction of some neurons varied in correlation with alternating brain states, indicating dynamics in the information processing that takes part within the NI. This dynamic information processing within the NI contributes to understanding stress and reward mechanisms, providing potential therapeutic avenues for stress-related disorders.
Funding: National Science Centre in Poland, UMO-2022/45/N/NZ4/03171.
Roszkowska J1, Drwięga G1 , Walczak M1, Mierniczek O1, Gorkowska M1, Pradel K1,2, Solecki W3, Błasiak T1
1Department of Neurophysiology and Chronobiology, Jagiellonian University, Cracow, Poland
2Institute for Systems Physiology, University of Cologne, Cologne, German
3Department of Neurobiology and Neuropsychology, Jagiellonian University, Cracow, Poland
Abstract: The brainstem nucleus incertus (NI) is involved in modulating stress response and aversive stimuli processing. It innervates ventral tegmental area (VTA) and rostromedial tegmental nucleus (RMTg) – the primary inhibitory input to the VTA dopaminergic (DA) neurons. In order to visualise the circuit’s anatomy, retrogradely transported viral vectors, carrying genes for fluorescent proteins, were unilaterally injected into the RMTg of Sprague-Dawley rats. The results revealed that, unlike the ipsilaterally innervated VTA, the RMTg receives bilateral innervation from the NI. The following electrophysiological recordings in urethane-anesthetised rats were preceded by injections of two viral vectors: one retrograde, carrying Cre recombinase gene, to the VTA or RMTg and another, carrying Cre-dependent genes for an excitatory opsin, which targeted the NI. The results revealed functional effects of NI-originating innervation on the two midbrain structures. Lastly, preceding behavioural experiments, retrograde viral vectors carrying YFP were injected bilaterally into the animals’ RMTg. Following a stress-induction procedure in operant conditioning chamber, a series of immunostaining was performed. The resulting data provide insight into the presumed overlap between c-fos-reactive NI subpopulation and the neurons innervating the RMTg. Altogether, this suggests that NI-originating innervation of midbrain structures forms a complex system involved in stress and reward processing.
Funding: National Science Centre: OPUS_17_2019/33/B/NZ4/03127, Preludium_21_2022/45/N/NZ4/03171
Patryk Sambak1,3, Anna Gugula1, Agata Szlaga1, Sylwia Drabik1,3, Gniewosz Drwiega1,3, Piotr Rywczak1, Aleksandra Trenk1, Angelika Kaleta1, Tomasz Blasiak1, Andrew L. Gundlach2, Anna Blasiak1
1Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland
2The Florey Institute of Neuroscience and Mental Health, and Florey Department of Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia
3Doctoral School of Exact and Natural Sciences, Jagiellonian University, Krakow, Poland
Abstract: Accurate discrimination between new and familiar stimulus is crucial for adaptive responses, with disruptions strongly linked to neuropsychiatric disorders. Stress significantly impairs novelty related responses. Thus, we investigated how the stress-sensitive brainstem nucleus incertus (NI), which is the primary source of the neuropeptide relaxin-3 (RLN3), may control the interpeduncular nucleus (IPN), which is crucial in signaling novelty preference. Multiplex in situ hybridization revealed RXFP3 mRNA expression in dopaminergic D1 and cholinergic α5 receptor mRNA-expressing IPN neurons, implicated in novelty and familiarity signaling, respectively. Patch-clamp recordings combined with optogenetic stimulation showed direct NI originating, inhibitory innervation of IPN in male rats. Viral tract-tracing unveiled NI-IPN neurons projecting to regions controlling stress and novelty related behaviours. Behavioral studies showed that chemogenetic activation of NI-IPN axis increased exploration time and reduced grooming in open field test. Our findings suggest NI involvement in stress-related novelty preference control, and sheds light on the neuronal mechanisms underlying stress-related impaired novelty preference.
Funding: National Science Centre Poland UMO-2018/30/E/NZ4/00687, UMO-2023/49/B/NZ4/01885; Jagiellonian University U1U/W18/NO/28.40.
Aleksandra Koszałka1,2 , Klaudia Lustyk1 , Henryk Marona3, Karolina Pytka1
1Department of Pharmacodynamics, Faculty of Pharmacy, Jagiellonian University Medical College, Krakow, Poland
2Doctoral School of Medical and Health Sciences, Jagiellonian University Medical College, Poland
3Chair of Organic Chemistry, Department of Bioorganic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Poland
Abstract: Anxiety disorders stand among the most widespread mental health challenges, impacting approximately 4% of the worldwide population. Central to this is stress, which, especially when coupled with ineffective coping mechanisms, can augment anxiety levels, and compromise therapeutic outcomes. Given that women experience anxiety more frequently than men, biological sex should be regarded as a potential variable in effective treatment development. Therefore, our study examines the effects of HBK-15, a novel multimodal compound, on anxiety-like behaviors and acute stress-coping in female mice. Animals were given a single dose of HBK-15 and then underwent the four-plate test to measure its anxiolytic-like properties. Another group of mice underwent the forced swim test to evaluate how HBK-15 influenced the stress-coping strategy. At a dose of 0.15 mg/kg, HBK-15 significantly elevated the number of punished crossings in the four-plate test. Meanwhile, administration of 0.3 mg/kg resulted in a 20% decrease in immobility time during the forced swim test. Our results demonstrate that HBK-15 exerts an anxiolytic-like effect on female mice. Moreover, it promotes an active coping strategy following acute stress exposure. Although in both cases the effect was observed in a narrow dose range. Such a dual-action profile holds promise for advancing therapeutic approaches in anxiety treatment.
Funding: This study has been conducted as part of a research project financed by the National Science Centre, Poland (grant number 2019/34/E/NZ7/00454).
Paulina Dudzik1, Nathalie Idlin1, Diego Pascal Garcia2, Henryk Marona3, Dorota Żelaszczyk3, Jacek Sapa1, Karolina Pytka1, Klaudia Lustyk1
1Department of Pharmacodynamics, Faculty of Pharmacy, Jagiellonian University Medical College, Krakow, Poland
2Faculty of Pharmacy, University of Valencia, Valencia, Spain
3Department of Bioorganic Chemistry, Chair of Organic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Krakow, Poland
Abstract: Schizophrenia is a severe mental disorder defined by disturbances in thought processing and perception of reality, leading to a lifelong disability. Although conventional antipsychotic therapy can alleviate symptoms, such as hallucinations and delusions, addressing memory deficits remains a challenge. Therefore, there is an urgent need to develop novel antipsychotics with additional procognitive properties. Building on our previous study on HBK-10, a multimodal compound targeting the D2 and 5-HT1A receptors, we aimed to further explore its antipsychotic-like and antiamnesic activity.
First, we conducted the MK-801- and amphetamine-induced hyperlocomotion tests. Subsequently, we performed an object recognition test with test phase 15 min and 24 hours after the administration of compounds, using MK-801 to impair memory. In all experiments, we employed male CD-1 mice.
HBK-10 at higher tested doses showed antipsychotic-like activity, reducing hyperlocomotion caused by both MK-801 and amphetamine. Additionally, at doses of 1.25, 5 and 10 mg/kg, it reversed MK-801-induced short-term recognition memory impairments. Considering its antiamnesic effect on long-term recognition memory, only the dose of 0.625 was active. Our study reveals HBK-10's antipsychotic-like and antiamnesic properties, encouraging further research.
Funding: This study was financed by the Jagiellonian University Medical College (grant number N42/DBS/000288 and N42/DBS/000329).
Anurag Soren1, Klaudia Lustyk1, Kinga Sałaciak1, Dorota Żelaszczyk2, Henryk Marona2, Karolina Pytka1
1Department of Pharmacodynamics, Faculty of Pharmacy, Jagiellonian University Medical College, Cracow, Poland
2Department of Bioorganic Chemistry, Chair of Organic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Cracow, Poland
Abstract: Cognitive deficits of varying severity have often been observed across neuropsychiatric conditions. Memory impairments significantly deteriorate the daily functioning of the patients and may affect the effectiveness of the used medications. Hence, there is an urgent need for therapies that improve cognitive performance.
Our previous research has identified HBK-10, an antagonist of 5-HT1A and D2 receptors, as a promising compound with demonstrated antidepressant-like properties in mice.
To further investigate, our study concentrated on determining the effect of HBK-10 on short-term, intermediate, and long-term recognition memory using the object recognition test in CD-1 mice.
Our findings indicate that HBK-10, within a dosage range of 0.625 to 10 mg/kg, does not significantly impair short- and long-term recognition memory in mice. However, at doses of 0.625 and 5 mg/kg, it adversely affects intermediate recognition memory.
This calls for more research to thoroughly explore the full potential of HBK-10.
Funding: This study was financed by the Jagiellonian University Medical College (grant number N42/DBS/000288).
Vilius Kasparavičius1, Simonas Kutanovas2, Neringa Daugelavičienė2, Urtė Neniškytė1,2
1Institute of Bioscience, Life Sciences Center, Vilnius University, Vilnius, Lithuania
2VU LSC-EMBL Partnership for Genome Editing Technologies, Life Sciences Center, Vilnius University, Vilnius, Lithuania
Abstract: Genome editing is making its way towards therapeutic applications. However, genome editing tools are usually researched using cancerous, undifferentiated cell lines as research models, which do not accurately represent living organisms. Widely used neuronal model SH-SY5Y neuroblastoma cell line is commonly used in undifferentiated state that is not representative of mature neurons. Differentiated SH-SY5Y neural-like cells are distinguished from undifferentiated ones by their neural marker expression, neurite formation and halt of proliferation. We established a dual-reporter SH-SY5Y cell line expressing EGFP and mRuby2, which could be used for gene editing tool screening, evaluating tool efficiency in differentiated and undifferentiated cell states at the single cell level. The dual-reporter system was generated by template knock-in via CRISPR-mediated HDR into the safe harbour AAV1 locus of SH-SY5Y cells. Donor template and CRISPR-Cas were transfected via lipofection. In the template plasmid, mRuby2 was positioned near its own promoter, for easy visualization upon successful transfection. EGFP, along with the puromycin resistance gene, was driven by the cell’s endogenous promoter, which ensured EGFP expression only after a successful integration into the genome. Furthermore, we assessed SH-SY5Y differentiation conditions for neural-like cell generation.
Anna Alwani, Piotr Chmielarz
Department of Brain Biochemistry, Maj Institute of Pharmacology PAS, Kraków, Poland
Abstract: Brainstem is a midline structure comprised of numerous nuclei. Notable among these nuclei are the locus coeruleus (LC), rich in noradrenergic neurons, and the nuclei raphes (NR), predominantly composed of serotonergic neurons. These neurons are linked with the regulation of mood, arousal, stress responses, sleep-wake cycles, and cognitive functions. Despite their significance, they remain largely understudied in the context of Parkinson’s Disease (PD), despite being among the major cell groups affected by degeneration in this condition. Establishing methodology for isolating and culturing primary noradrenergic and serotonergic neurons holds promise for advancing our understanding of their involvement in PD etiology.
Brainstem was isolated from embryonic mice at days 15-18 and specific fragments located within the fourth ventricle were excised followed by a cell dissociation procedure and plated onto polyornithine-coated 96-well plates in Neurobasal medium with B-27 supplement. After 7 or 14 days, the cells were fixed with 4% PFA and stained by immunofluorescence method to visualize a noradrenergic (NET/DBH marker) or serotonergic neurons (TPH marker).
Fluorescent imaging confirmed successful culturing of primary noradrenergic and serotonergic neuronal populations for prolonged periods of time.
Established protocols hold significant potential for investigating Parkinson's Disease-related pathology across diverse neuronal types, opening new avenues for further research exploration.
Funding: This work was supported by the statutory funds of the Maj Institute of Pharmacology, PAS, Poland
Fionä Caratis1,2, Mikołaj Opiełka1, Martin Hausmann3, Maria Velasco-Estevez4, Bartłomiej Rojek5, Cheryl de Vallière3, Klaus Seuwen3, Gerhard Rogler3, Bartosz Karaszewski1,5, Aleksandra Rutkowska1,2
1Brain Diseases Centre, Medical University of Gdansk, Gdansk, Poland
2Department of Anatomy and Neurobiology, Medical University of Gdansk, Gdansk, Poland
3Department of Gastroenterology and Hepatology, University Hospital Zurich, University of Zurich, Zurich, Switzerland
4H12O-CNIO Hematological Malignancies Group, Clinical Research Unit, Centro Nacional de Investigaciones Oncologicas (CNIO), Madrid, Spain
5Department of Adult Neurology, Medical University of Gdansk & University Clinical Centre, Gdansk, Poland
Abstract: Acidosis is a distinctive feature of demyelinating lesions in multiple sclerosis (MS) in the central nervous system (CNS). The cellular response to acidic pH is primarily regulated by a family of G protein-coupled proton-sensing receptors, including OGR1, GPR4, and TDAG8, which are inactive under alkaline conditions and maximally activated in an acidic environment. Genome-wide association studies have linked a locus within the TDAG8 gene to autoimmune diseases, including MS. We found an upregulation of TDAG8 expression in MS plaques, distinguishing it from GPR4 or OGR1. This led to a detailed exploration of TDAG8 expression and function in oligodendrocytes using in vitro and in vivo models in mice and humans. Surprisingly, we observed significant upregulation of TDAG8 in human MO3.13 oligodendrocytes during maturation and in response to acidic conditions. However, its deficiency did not impact normal myelination in the mouse CNS, and TDAG8 expression remained unchanged under demyelinating conditions in mouse organotypic cerebellar slices. Notably, we found no expression of TDAG8 in primary mouse oligodendrocyte progenitor cells (OPCs), in contrast to its presence in primary human OPCs. These findings highlight substantial species differences in the expression of the proton-sensing receptor TDAG8 in OPCs, underscoring the limitations of the models employed in comprehending the role of TDAG8 in myelination. Despite these model-specific constraints, our results suggest a potential role of TDAG8 in human myelination, implicating its involvement in the pathophysiology of MS, and provide valuable insights into the judicious selection of models for future scientific investigations.
Funding: This project received funding from the National Science Centre, Poland, grant registration number: 2019/33/B/NZ4/03000 (AR)
Sawicka Aleksandra1, Harackiewicz Oliwia1, Świątek Grzegorz1, Grembecka Beata1, Majkutewicz Irena1, Pogorzelska Karolina1, Szczepaniak Magdalena1, Honkisz-Orzechowska Ewelina2, Wrona Danuta1
1Department of Animal and Human Physiology, Faculty of Biology, University of Gdańsk, Gdańsk, Poland
2Departament of Technology and Biotechnology of Drugs, Faculty of Pharmacy, Jagiellonian University in Cracow Medical College, Cracow, Poland
Abstract:
Background: Recent studies demonstrated that Delta9-Tetrahydrocannabinol (THC) prevents neurodegenerative processes occurring in animal models of Alzheimer’s disease (AD) and protects from inflammation-induced cognitive damage in old mice.
Methods: In the present study we investigated an influence of intraperitoneal injections of THC at a dose of 5 mg/kg b.w. for 7 consecutive days on spatial memory, measured as a latency to reach the platform and total swimming distance before escaping onto the visible platform in Morris water maze test (MWM) in rats with the streptozotocin (STZ)-induced inflammation model of Alzheimer’s disease (AD).
Results: In the STZTHC group a significantly shorter (p<0.05) total distance (25947.23±2968.37 cm; mean±SD) rather than in the STZTHC solvent group (32016±3580.99 cm) was observed. As compared to the control VEHTHC solvent group, both the latency to reach the platform (2.58 ± 1.17 s) and total distance (22144.88 ± 3025.34 cm) were significantly (p<0.05) longer in the STZTHC animals. Moreover, increased total distance (p<0.05) but not latency in the STZTHC solvent rather than VEHTHC solvent (22144.88±3025.34 cm) animals was noticed.
Conclusions: The seven-day THC (5 mg/kg b.w.) injections could improve spatial memory disorders, as indicated by decreased total swimming distance in the MWM in rats with the inflammation-induced AD model.
Funding: This work was supported by the grant no. 2021/43/D/NZ3/01440 (Sonata) from the National Science Centre, Poland.
Bernadeta A. Pietrzak-Wawrzyńska, Agnieszka Wnuk, Karolina Przepiórska-Drońska, Andrzej Łach, Małgorzata Kajta
Maj Institute of Pharmacology, Polish Academy of Sciences, Laboratory of Neuropharmacology and Epigenetics, Krakow, Poland
Abstract: Alzheimer's disease (AD) presents a serious challenge in neurodegenerative disorders due to the accumulation of amyloid-β (Aβ), leading to neuronal dysfunction and cognitive decline. Estrogen receptors (ERs) have emerged as potent mediators of neuroprotection against AD, with non-nuclear ER activation considered safer than nuclear activation. This study investigates PaPE-1, a compound activating non-nuclear ER pathways, for its neuroprotective effects in AD. To model AD, we used mouse primary neocortical cell cultures exposed to Aβ. PaPE-1 was applied with 24 hour-delay and the treatment lasted for 6 hours. The presence of extracellular Aβ aggregates and changes in the expression of AD-related markers (Rbfox, Ache, Apoe, Chat, Ngrn) confirmed the adequacy of our model. We showed that Aβ‑induced cell death involves apoptosis‑specific pathways. PaPE-1 influenced the expression of AD-related markers and decreased apoptotic cell death in terms of caspase-3 activity. PaPE-1 exerts regulatory control over apoptosis at different stages, including DNA methylation, mRNA expression, and protein level of Bcl2/BCL2 and Bax/BAX. This study highlights PaPE-1's neuroprotective potential and unveils its novel mechanisms of action, supporting the therapeutic promise of targeting non-nuclear ER signaling pathways in developing innovative AD therapies.
Funding: National Science Centre of Poland, grant number 2020/39/NZ7/00974
Natalia Siwecka, Grzegorz Galita, Zuzanna Granek, Wojciech Wiese, Wioletta Rozpędek-Kamińska, Ireneusz Majsterek
Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, Lodz, Poland
Abstract:
Introduction: Parkinson’s disease (PD) is a neurodegenerative disorder caused by death of dopaminergic neurons. The main molecular mechanism underlying PD is oxidative stress, which may be induced by pro-oxidants like 6-hydroxydopamine (6-OHDA). JNK is a major pro-apoptotic kinase involved in pathogenesis of many diseases, and it was also reported to have a key role in PD.
Aim: The present study aimed to investigate the effect of pharmacological JNK inhibition in cellular model of PD.
Methods: The study was conducted on SH-SY5Y cells differentiated with retinoic acid. Neurodegeneration was induced by treatment with 6-OHDA. Cells were treated with JNK inhibitor V either before or after 6-OHDA-induced damage. The cell viability was measured by XTT assay, genotoxicity – by comet assay, and the mRNA expression level of specific genes – by RT-qPCR analysis.
Results: Inhibition of JNK significantly improved cell viability, even when the inhibitor was applied after 6-OHDA-induced damage. The protective effect of JNK inhibition was also observed against 6-OHDA genotoxicity. Gene expression analysis revealed a significant downregulation of MAPK10, XBP1 and DDIT3 by JNK inhibitor.
Conclusions: The results obtained indicate neuroprotective effects of pharmacological JNK inhibition against 6-OHDA-induced damage. Thus, JNK inhibitors could potentially be applied for the selective treatment of PD.
Funding: Grant no. 2021/43/O/NZ5/02068 from the National Science Centre, Poland.
Zuzanna Granek, Natalia Siwecka, Wioletta Rozpędek-Kamińska, Grzegorz Galita, Ireneusz Majsterek
Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, Poland
Abstract:
INTRODUCTION
Neurodegeneration in Parkinson’s disease (PD) is associated with accumulation of α-synuclein and induction of Endoplasmic Reticulum (ER) stress in dopaminergic neurons, which activates the PERK branch of the UPR signalling pathway. PERK orchestrates neural cell apoptosis via upregulation of specific pro-apoptotic genes, which makes it a perfect target for development of novel treatment strategies against PD.
OBJECTIVES
The primary objective of the study was to evaluate the effectiveness of the selected small-molecule PERK inhibitor LDN-87357 (LDN) in PD in vitro model.
METHODS
XTT assay was used for the cytotoxicity analysis. SH-SY5Y cells were exposed to LDN at 0.75-100µM, 50µM 0.1% DMSO (solvent for LDN) and 500nM ER stressor, thapsigargin (Th). Cells treated with Th only served as a positive control, whereas untreated cells as a negative control. qPCR was performed to assess the mRNA expression levels of specific ER stress related pro-apoptotic genes including DDIT3, BAX, ATF4 and GADD34 and anti-apoptotic gene BCL‐2. Cells were treated with LDN at 0.75 or 50µM, 0.1% DMSO, 500nM Th or with LDN+Th. Untreated cells constituted a negative control.
RESULTS
XTT test demonstrated no cytotoxic effect of investigated PERK inhibitor towards SH SY5Y cells. Importantly, the viability of cells exposed to Th was significantly increased upon pretreatment with LDN. qPCR revealed a significant decrease in the expression of pro-apoptotic genes, and an increase in anti-apoptotic genes expression in SH SY5Y cells with induced ER stress conditions, as compared to only Th-treated cells.
CONCLUSIONS
To effectively treat PD, new drugs which directly target the molecular pathways involved in pathophysiology of the disease need to be developed. We may assume that targeting PERK via small-molecule inhibitors, such as LDN-87357, may contribute to development of a novel targeted therapy against neurodegenerative diseases, that would provide neuroprotection and have no cytotoxic effect.
Funding: PRELUDIUM BIS 3 no. 2021/43/O/NZ5/02068, National Science Centre, Poland
K. Maziarz1, M. Jankowska-Kiełtyka1, J. Barut1, G. Burda1, P. Chmielarz1
1Maj Institute of Pharmacology Polish Academy of Sciences, Department of Brain Biochemistry, Kraków, Poland
Abstract: Parkinson’s Disease (PD) is neurodegenerative disorder which motor symptoms are connected with progressive loss of dopamine neurons. Another sign of PD is Lewy Pathology which formation and transmission are linked with alpha-synuclein’s (a-syn) prion properties. We have previously demonstrated that activation of Akt and Src pathways by neurotrophic factors reduce accumulation of a-syn.
Here our goal was to determine if activation of those pathways by Ghrelin Receptor agonist MK-0677 will be also protective.
Mice injected into striatum with prion a-syn fibrils (PFFs) were treated with saline (SAL) or MK-0677 at different doses for either 1 or 4 months, followed by behavioral tests and immunostaining for markers of dopamine neurons and a-syn aggregates.
After 1 month PFF+SAL mice exhibited impaired motor coordination on multiple static rod test which was reversed by MK-0677 at 1mg/kg and 2mg/kg. PFF+SAL group had decreased number of TH+ neurons, rescued by MK-0677 at 0.2mg/kg, 1mg/kg and 2mg/kg. Number of a-syn aggregates in TH+ cells increased in 0.2mg/kg dose, decreased in 2mg/kg. After 4 months MK-0677 at 2mg/kg clearly decreased amount of a-syn aggregates in TH+ cells.
Our results strongly support further investigations of GHS-R agonist for treatment of PD at early stages.
Funding: This research was funded by National Science Centre, Poland, grant number 2019/35/D/NZ7/03200 (Sonata 15)
Julia Jarco, Natalia Malek
Department of Chemical Biology and Bioimaging, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland
Abstract: The GPR84 receptor, an orphan G-protein-coupled receptor, is expressed in various immune cells, including microglia, the resident macrophages of the central nervous system (CNS). Microglia play a significant role in the pathophysiology of CNS disorders. While the activation of GPR84 is associated with the regulation of inflammatory processes, the precise mechanism remains elusive. As microglial death and its consequences serve as a means of communication within the CNS, our study aims to investigate the effects of GPR84 activation on the regulation of pyroptosis and apoptosis pathways in human microglial cells. Employing a pharmacological approach, we analysed gene expression associated with pyroptosis and apoptosis using RT-qPCR. We observed an increase in GPR84 expression during microglial apoptosis, whereas its expression decreased during pyroptotic cell death. Interestingly, treatment with DL175 (a GPR84 agonist) resulted in increased expression of the main pyroptotic factor, NLRP3, in control cells, but decreased expression during pyroptosis. Additionally, altered expression of caspase genes was noted in HMC3 cells following GPR84 activation. These findings suggest that GPR84 may regulate microglial cell death by modulating pyroptosis and apoptosis through distinct mechanisms, contributing to our understanding of its role in CNS immune response regulation.
Funding: Supported by National Science Center, Poland grant OPUS 2020/37/B/NZ7/03411.
Emilija Napieralska, Joanna Kula, Katarzyna Zofia Kuter
Department of Neuropsychopharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences, Cracow, Poland
Abstract: Astrocytes – star-shaped glial cells support neurons structurally and energetically, but also coordinate their functions in e.g. memory or compensatory processes. Astrocytes are naturally heterogeneous and vary in terms of localization, morphology, protein expression and activation state. In this study, we documented those differences in cerebral cortex, striatum and substantia nigra. To observe this heterogeneity in vivo, immunohistochemically stained brain sections were examined in a rat model for selective, prolonged astrocyte dysfunction by constant infusion of fluorocitrate and in a 6-OHDA dopaminergic system lesion model. Astrocyte-specific GFAP, glutamine synthetase and s100β protein expression were detected. In astrocyte depletion model, activated astroglia surrounded lesioned area probably creating a „glial scar”. In contrast, neuronal degeneration did not affect astrocytic markers. Regional differences in cell density and shape were visible in particular markers. Marker protein expression showed that astrocytes tailor their morphology to specific brain parts, most probably to perform region and neuron type-dependent functions. Observed variability also hints at functional flexibility and indicates necessity to co-detect multiple markers to better understand molecular context of changes. Astrocyte-specific markers have already been put into discussion as perspective tools for neurodegenerative disease diagnostics.
Funding: National Science Centre grants 2017/27/B/NZ7/00289 and statutory funds of IP PAS.
Justyna Barut, Michał Wilczkowski, Katarzyna Maziarz, Katarzyna Rafa-Zabłocka, Piotr Chmielarz, Grzegorz Kreiner
Department of Brain Biochemistry, Maj Institute of Pharmacology, Polish Academy of Sciences, Kraków, Poland
Abstract: Neuroinflammation and glial reactivity contribute to dopaminergic neuron degeneration in Parkinson's disease. Noradrenergic enhancement shows promise as a neuroprotective strategy against dopaminergic cell loss, partly by reducing glial activation. We investigated whether noradrenergic transmission is mediated by astrocytes.
We induced a reactive phenotype in primary mouse astrocytes using IL-1α-TNFβ or IL-1β-TNFβ treatment, leading to characteristic changes: altered morphology, decreased GFAP expression, increased proliferation, and Nf-kB translocation to the nucleus. Cytokine stimulation increased monocarboxylate transporter expression, indicating metabolic changes, confirmed in seahorse studies. In midbrain neuronal cultures, cytokine treatment didn't affect TH+ dopaminergic neuron numbers, but co-culturing with astrocytes led to dose-dependent TH+ neuron reduction, mitigated by α-adrenergic receptor agonists. In mouse models mimicking Parkinson's disease (PD), progressive noradrenergic cell degeneration and phenotypic changes occurred without TH+ cell loss in the SN/VTA but with increased neurodegenerative markers, GFAP expression, and astrocytic chemokines. Elevated genes in the SN included among other Sgk1, an astrocytic GR-dependent gene.
Our results provide data on the possible negative influence of noradrenergic degeneration on SN/VTA, confirming the neuroprotective role of noradrenaline, possibly also mediated by astrocytes. Primary astrocytes after IL-1α-TNFβ or IL-1β-TNFβ treatment acquire reactivity and are a useful model for in vitro research.
Funding: 2019/35/N/NZ7/03300 financed by the Polish National Science Center
Carla Ramon-Duaso, Júlia Pinho and Arnau Busquets-Garcia
Hospital del Mar Research Institute, Cell-type mechanisms in normal and pathological behavior, Barcelona, Spain
Abstract: Astrocytes are implicated in several cellular and synaptic mechanisms that play a crucial role in different pathophysiological and behavioral phenotypes linked to Alzheimer's disease (AD). Our aim is to elucidate the contribution of astrocytes in different AD phenotypes, opening a window to new treatments against cognitive deficits in AD.
In this study, we used male and female APP/PS1 mice as a model of AD. First, we set up a behavioral analysis combining EzTrack® and DeepLabCut™ with handmade Python scripts. Notably, we observed sex- and genotype-dependent effects in different cognitive domains. Second, we used in-vivo fiber photometry to investigate whether astrocyte calcium dynamics are linked with memory alterations. Animals at five-months-old were operated and infused with pZac2.1 gfaABC1D-cyto-GCaMP6f virus in dorsal and ventral hippocampus. After 4 weeks, animals performed the behavioural paradigms in a within-subjects design (novel object recognition and light-tone sensory preconditioning). Our preliminary data suggest sex- and/or genotype-dependent differences in hippocampal calcium dynamics at different phases of the behavioral tasks.
Overall, this data suggests a direct link between astroglial calcium dynamics and the behavioral differences found in an AD’ animal model, which should be further confirmed with causal experiments.
Magdalena Siwarga, Milena Damulewicz
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Cracow, Poland
Abstract: α-Synuclein (α-Syn) is a protein encoded by SNCA gene, involved in the regulation of synaptic vesicle trafficking and neurotransmitter release. However, mutated form of α-Syn has high ability to aggregation and it is a major component of protein inclusions known as Lewy bodies, which are hallmarks of synucleinopathies including Parkinson's disease (PD). In our studies we used Drosophila melanogaster as a model of familial PD linked with mutation of SNCA. Insects do not express synuclein, however using transgenic flies we were able to prepare flies with expression of human wild-type and E46K- mutated α-synuclein in glial cells and neurons, respectively. Previous research, has shown that E46K α-synuclein caused remarkable climbing defects, as well as reduced survivorship. However, there is still much that remains unknown about the role of α-synuclein. In the present project, we compared the effects of expression of wild-type and E46K- mutated α-synuclein on behaviour of young and old flies.
Kamila Zientara, Milena Damulewicz
Department of Cell Biology and Imaging, Jagiellonian University, Cracow, Poland
Abstract: Parkinson's disease (PD) is the second most common neurodegenerative disorder. One of the early onset form of PD is associated with mutations in the park gene, which is constitutive among species. PARKIN starts the process of ubiquitination and degradation of damaged mitochondria on the pathway called mitophagy, which disruption leads to increase oxidative stress which causes cell death. There are strong evidences about connection between PD progression and disruptions of the circadian machinery. One of the main models in chronobiology is Drosophila melanogaster due to its simplified clock system consisting of 150 neurons divided into 7 groups, including the sLNv neurons that release PDF (Pigment Dispersing Factor) neurotransmitter crucial for circadian activity pattern. In this study, we tried to investigate how ageing contributes to circadian disruptions caused by park mutation. First, we examined the locomotor activity and sleep profiles of young and 30-day-old park mutants. Then, we isolated brains at two time points, at the beginning of the day and the night, and using anti-PDF immunostaining we visualized sLNv neurons to analyse daily axonal plasticity. Young PD model flies exhibited reduced sleep time and increased sleep fragmentation, along with disrupted rhythmicity in sLNv neuron axons complexity. Compared to those results, old flies showed decreased activity amplitude during the day. We also observed changes in the branching pattern of sLNv neurons.
Overall, our results showed that PD model flies have disrupted circadian clock and observed changes increase with age.
Kornel Szypulski, Milena Damulewicz
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Cracow, Poland
Abstract: Paracetamol is a popular over-the-counter analgesic and antipyretic drug. Despite being considered relatively safe it can have toxic effects via one of its metabolites called N-acetyl-p-benzoquinone imine (NAPQI). NAPQI depletes reduced form of glutathione and binds to cellular macromolecules resulting in oxidative stress and cytotoxicity. In our study we investigated the effect of paracetamol on Canton-S (wild type control), w1118 (genetic background control) and park1 (mutation in park gene, model of early-onset autosomal recessive Parkinson’s disease (PD)) flies. For park1 strain experiments were performed separately on individuals that were homo- or heterozygous for the mutation. Young flies were exposed to 100mM paracetamol for 24h and then collected for gene expression analysis or transferred back on the standard cornmeal medium for survival assay. Exposure to paracetamol increased mortality of homozygous mutants, but did not affect heterozygotes or control flies. There were also changes in the expression levels of gstd2 (glutathione S-transferase D1) gene, genes related to oxidative stress response: sod1 (superoxide dismutase 1) and trxr1 (thioredoxin reductase 1) and dopamine-pathway-related genes: dat (dopamine transporter) and ple (tyrosine hydroxylase). Since the toxic effects of paracetamol are especially detrimental for PD-modelling Drosophila strain these results suggest that mutation in park gene may increase susceptibility to toxic effects of otherwise safe drug.
Helena Kotlicka, Emil Wawak, Mateusz Zaborski, Milena Damulewicz
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Cracow, Poland
Abstract: The Earth rotation around the Sun provides dramatic changes in light conditions during the day and night. Most of organisms is light-sensitive, and keep diurnal or nocturnal life style. Light has both, positive and negative effects on an organism; from being a regulator of circadian rhythms to being a factor in ROS production, which can lead i.e. retina degeneration.. Drosophila melanogaster is a great model for studying molecular and cellular mechanisms of human diseases such as Parkinson’s disease. Short lifespan and simple behavior causes that this organism is commonly used in neurobiology of aging.
In our study we tried to investigate how different light conditions affect survival of fruit flies. We used males coming from different strains: wildtype CantonS, white mutants and park mutants (both homozygous and heterozygous).
We used three different light settings; 12h of light:12h of darkness (LD12:12), constant darkness (DD) and 12h of light:12h of dim light (L-dim). The last conditions simulated light pollution in the dark phase observed in urbanized areas. Experiment started with 30 males per genotype. Survival was checked every day starting from emergence until the death of the last fly in the group. Experiment was repeated three times. We observed statistically significant differences between flies kept under different light regimes. Our results suggest that presence of light may shorten the lifespan, which may be connected with increased oxidative stress levels.
Kornelia Tryzno, Jakub Frydrych, Ewa Trojan and Agnieszka Basta-Kaim
Laboratory of Immunoendocrinology, Department of Experimental Neuroendocrinology, Maj Institute of Pharmacology, Polish Academy of Sciences, Kraków, Poland
Abstract: Despite extensive research over many years, there is still no consensus on the etiology and pathogenesis of Alzheimer's disease (AD), making it difficult to establish effective treatment and/or prevention strategies. Insulin has been observed to be significantly involved in cell survival, metabolic function and neuronal plasticity. In the present study, the age-dependent expression of insulin (Ins1) and its receptor (Insr) in the frontal cortex and hippocampus of control and transgenic APP NL-F/NL-F mice was examined using real-time polymerase chain reaction. Due to the involvement of adaptor proteins in the insulin pathway, the expression of Irs1, Irs2, Shc-1 and Grb2 was also investigated. It was found that in the AD model studied, the mutation significantly affects the expression patterns of genes involved in insulin signalling, with different effects depending on the brain region studied and the age of the animals. These changes are dynamic and may play a key role in AD pathology, especially given their occurrence early in animal development. Further exploration of the mechanisms underlying these regulatory dysfunctions may lead to the development of new therapies or preventive measures for AD.
Funding: Funded by the National Science Centre, Grant No: 2021/43/B/NZ4/01133 Task 5 and partially statutory founds of the Department of Experimental Neuroendocrinology, Maj Institute of Pharmacology PAS
Monika Mitka, Jolanta Górska-Andrzejak
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research,
Jagiellonian University, Kraków, Poland
Abstract: Exosomes have been widely recognised as communication vesicles that transfer bioactive proteins and RNAs between cells under normal and pathophysiological conditions. In this study, we investigate at the cellular and behavioural levels whether this type of cellular communication is subject to circadian regulation in the fruit fly Drosophila melanogaster.
We used transgenic flies with the targeted expression of GFP-tagged mammalian tetraspanin (Tsp), CD63, to examine its daily expression in neurons and glial cells. We also examined the prominent output of Drosophila circadian clock, the circadian rhythm of locomotor activity, in flies with silenced expression of Tsp42Eg and Tsp42Ee in neurons and glial cells of the circadian network. For this study we used confocal microscopy and the Drosophila Activity Monitoring System (DAMS).
We found daily changes in CD63 expression in glial cells of Drosophila brain. We also detected changes in the daily and circadian pattern of locomotor activity of flies expressing CD63 (with overexpression of tetraspanins) and flies with silenced expression of Tsp42Eg and Tsp42Ee. This implies circadian regulation of this type of cellular communication and suggests that exosomal communication may be an important part of the functioning of the circadian clock/network.
Funding: This work was supported by the Department of Cell Biology and Imaging of the Institute of Zoology and Biomedical Research (Grant number N18/DBS/000015).
Lysikova Terezia, Leskova Majdova Katarina, Racay Peter, Kaplan Peter, Tatarkova Zuzana
Department of Medical Biochemistry, Jessenius Faculty of Medicine in Martin, Comenius University Bratislava, Slovakia
Abstract: Ischemic preconditioning (IPC) triggers endogenous neuroprotection to defend against subsequent, more severe ischemia. IPC takes advantage of brain plasticity for its neuroprotective purposes, especially for energy and redox homeostasis. Mitochondria sense and respond to diverse harmful stresses, aiding cells in adaptation and resilience, with proteins being a primary focus of their defensive mechanisms. In our study, 33 hippocampal and 15 cortical mitochondrial proteins exhibited different susceptibility to IPC and ischemia-reperfusion (IR). IPC upregulated glyceraldehyde-3P dehydrogenase and glutamine synthetase in the hippocampus while downregulating isocitrate and succinate dehydrogenase. Peroxiredoxins (Prx) stand out among antioxidant enzymes, with Prx5 and Prx6 levels declining post-IPC but escalating after IR, notably in CA1 and CA3 hippocampal regions. IPC elevated ATP synthase subunit beta and Mn superoxidase dismutase in cortical mitochondria, alongside enhanced ATP synthase, glutathione peroxidase and glutathione reductase activities. Conversely, IR downregulated cytochrome c reductase aconitate hydratase and pyruvate dehydrogenase. Hippocampal proteins exhibit ROS sensitivity, particularly impacting Prx pathways, leading to energy metabolism dysregulation post-IPC. In contrast, the cortex maintains mitochondrial antioxidant availability during cerebral IR injury, indicating region-specific responses to IPC and IR, crucial for targeted neuroprotective strategies.
Funding: This research was supported by grant VEGA 1/0004/19.
Dorota Malicka, Bozena Kuzniewska, Marta Magnowska, Magdalena Dziembowska
Laboratory of Molecular Neurobiology, Department of Animal Physiology, Institute of Functional Biology and Ecology, Faculty of Biology, University of Warsaw
Abstract: Neuronal cells depend on mitochondrial activity to maintain membrane excitability, neurotransmission, and synaptic plasticity. Production of ROS can cause mitochondrial DNA damage. The most common mutation is the oxidation of guanine to 8-oxo guanine. The accumulation of mutations in mtDNA can lead to the impairment of mitochondria function. Fusion and fission determine the mitochondrial morphology and adapt it to the metabolic needs of the cells. Therefore, these two processes are crucial to optimize mitochondrial function.
TRAP1 is a mitochondrial chaperone protein that belongs to the HSP90 family. This protein is involved in protection against oxidative stress and regulation of mitochondrial dynamics. In a knock-in mouse model of Trap1 (p.Gln639Ter), the expression level of this protein is significantly reduced and the behavior of mouse mutants is affected. We isolated mitochondrial DNA from the brains of mutant and WT mice. MtDNA was sequenced to compare the amount of the mutations accumulation in different genotypes. Moreover, we isolated hippocampal and cortical mitochondria to investigate the levels and phosphorylation status of proteins involved in fission and fusion.
N. Ceccato1, M. Damulewicz2, G. Mazzotta1
1Department of Biology, University of Padua, Italy
2Department of Cell Biology and Imaging, Jagiellonian University, Krakow, Poland
Abstract: Circadian rhythms are daily fluctuations in behaviour and physiology, generated by internal timekeeping mechanisms and entrained by environmental cues. Our research focuses on the retina as it possesses a functional circadian clock mechanism and represents a hallmark for neurodegeneration and senescence. Using Drosophila melanogaster as a model organism, we aim to elucidate the involvement of the circadian clock in mitochondrial network within the retina. Indeed, these findings serve as a foundation for exploring the impact of circadian disruption on mitochondrial function in the context of neurodegeneration.
In this study we investigated daily changes in the number of mitochondria in specific photoreceptor type. We used transgenic flies in which mitochondrial-tagged fluorescent proteins were expressed under control of specific Rhodopsin type. Heads were collected at four time points and cryosections were prepared. Next, fluorescent signal was enhanced using immunostaining with anti-RFP antibodies. Finally, we analysed fluorescent intensity and the number of particles in the photoreceptor cell bodies and terminals, respectively. Our data suggest that mitochondria number in the retinal cells changes during the day.
Agata Małoburska, Milena Damulewicz
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Cracow, Poland
Abstract: Light intensity and composition changes throughout the day, which provides circadian clock synchronization. However, intense light induces production of reactive oxygen species which results in the physical and chemical changes in DNA structure. The retina is at the highest risk of light-induced DNA damages due to its phototransduction function and high level of metabolic processes. Although retina possess many protective mechanisms, they weaken with aging. In addition, the level of retinal DNA damage depends on the time during the day, when it is exposed to UV radiation, because the protective mechanism against the light is controlled by circadian clock.
In this study we investigated the consequences of UV radiation on Drosophila melanogaster retina in the morning (ZT1) and in the evening (ZT13). To investigate whether protective mechanisms change their effectiveness with age we used young (5-7 days) and old (30 days) wild type Canton-S flies. They were kept under the normal light condition (12h of light and 12h of darkness, LD 12:12) and decapitated at selected time points. Brains cryosections were immunostained using the anti-8-hydroxyguanosine primary antibodies to mark DNA breaks, which were then measured by comparing the reaction intensity using ImageJ. In young flies, more severe changes were observed after evening UV exposure. However, in old flies this difference between timepoints was not statistically significant, which suggest that age-related molecular changes in the circadian clock may affect retina vulnerability to light-dependent degeneration.
Kinga Skoczek, Milena Damulewicz
Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Kraków, Poland
Abstract: The aging process of the retina significantly impacts its structure, function, and susceptibility to degeneration, ultimately affecting overall health and visual function. Understanding this process offers valuable insights into age-related visual impairments and holds profound implications for neuroscience, given the close relationship between retinal and brain aging. Drosophila melanogaster, with its easy genetics, short lifespan, well-defined retinal structure, conserved molecular and genetic pathways, serves as an ideal model for such investigations.
A general decline in neuron function is associated with aging. This study examines whether specific changes occur also in photoreceptor neurons, particularly in Drosophila’s R1-R6 cells. These alterations may include molecular shifts affecting presynaptic proteins like Bruchpilot (BRP), crucial for calcium channels aggregation in the synaptic active zone and neurotransmitter release. Consequently, synaptic transmission relies on BRP, with decreased levels resulting in weaker transmission.
BRP expression was previously shown to be regulated by both, light and circadian clock, expressing distinct pattern – reaching maximum at the beginning of the day (ZT1) and night (ZT13). To investigate possible changes in the retina rhythmicity during aging we collected old flies at specific time points throughout the day (ZT1, ZT4, ZT13, ZT16). We cryosectioned heads and immunostained them with anti-BRP antibodies. Analysis of confocal images showed that aging retina maintains daily presynaptic protein profile observed in young flies. These results highlight the need for further investigation into the molecular mechanisms underlying age-related changes in Drosophila’s retina.
Patryk Mizia1, Bernard Kordas1, Julia Jarosławska2, Natalia Szyryńska3, Natalia
Ziółkowska3, Bogdan Lewczuk3, Judyta Juranek1
1Department of Human Physiology and Pathophysyology, Faculty of Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
2Department of Biological Functions of Food, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland
3Department of Histology and Embryology, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland
Abstract:
Background:
Diaphanous homolog 1 (Diaph1) is a protein crucial in neural tissue development, especially actin polymerization and microtubule formation. Lack of Diaph1 leads to neurodevelopmental disorders such as blindness, deafness, microcephaly, and seizures. The study aimed to compare the retinal structure of wild-type (WT) C57BL/6 mice with Diaph1 knockout (DKO) mice.
Methods:
Retinas from six mice (three WT, three DKO) were fixed in 2.5% glutaraldehyde and 1% paraformaldehyde, postfixed in 2% osmium tetroxide, dehydrated, and embedded in Epon 812. Morphometric analysis on semi-thin sections stained with toluidine blue was performed across five layers: Inner Plexiform Layer (IPL), Inner Nuclear Layer (INL), Outer Plexiform Layer (OPL), Outer Nuclear Layer (ONL), and Photoreceptor Layer (PL) using CaseViewer.
Results:
Statistical results highlighted significantly thinner OPL and PL in DKO mice (OPL: 11.94±3.62 µm DKO vs. 14.67±3.56 µm WT; PL: 44.80±4.85 µm DKO vs. 49.98±7.97 µm WT).
Conclusions:
The results highlighted Diaph1's essential role in retinal cilia formation. The OPL layer is a complex structure that consist of dendrites and axons of many neuronal cells, thus the impact of Diaph1 knockout requires further investigation.
Dorottya Molitor1, Alexandra Vaczy1, Edina Szabo1, Evelin Patko1, Balazs Meresz1, Dora Reglodi1, Tamas Atlasz1,2
1Department of Anatomy, MTA-PTE PACAP Research Team, University of Pecs Medical School, Pecs, Hungary
2Department of Sportbiology, Faculty of Sciences, University of Pecs, Pecs, Hungary
Abstract:
Background: A disorder associated with premature birth, called retinopathy of prematurity (ROP), is a neurovascular disease of the retina. The oxygen-induced retinopathy (OIR) is a well-established model of ROP characterized by vessel abnormalities. There are several factors which can result in premature birth, such as smoking during pregnancy. Our aim was to examine the vulnerabilities of maternal smoking on OIR.
Methods: C57BL/6 laboratory mice were used in this experiment. During the pregnancy mice had to smoke two times a day. To induce OIR, pups were exposed to hyperoxia from postnatal day (PD) 7-12 then returned to room air. On PD17 retinas were isolated. Isolectin GS-IB4 was used to label the endothelial cells of the retinas, then a computational tool was used for further quantitative analysis of the retinal vascular networks. Several antibodies were detected and quantified by western blotting of pooled retinas distributed by treatment groups.
Results: Our computational analysis of retinal vasculature showed quantitative changes in several parameters as well as in case of the protein expressions examined by western blotting compared to control conditions.
Conclusion: Based on our results we showed that maternal smoking caused a greater degree of retinal damage in ROP.
Funding: ÚNKP-22-3-I-PTE-1660; FK129190, K135457; National Brain Research Program NAP2017-1.2.1-NKP-2017-00002; MTA-TKI-14016; PTE AOK-TANDEM; GINOP-2.3.2-15-2016-00050 “PEPSYS”; EFOP-3.6.2-16-2017-00008; “The role of neuroinflammation in neurodegeneration: from molecules to clinics”; and Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary: 20765/3/2018/FEKUTSTRAT, 2020-4.1.1-TKP2020—FIKP III. Project No. TKP2020-IKA-08 has been implemented with the support provided from the National Research, Development and Innovation Fund of Hungary, financed under the 2020-4.1.1-TKP2020 funding scheme.
Li Lina1, Evelin Patko1, Andras Varga1, Diana Denes1, Dorottya Molitor1, Edina Szabo1, Balazs Meresz1, Dora Reglodi1, Alexandra Vaczy1, Tamas Atlasz1,2
1Department of Anatomy, HUN-REN-PTE PACAP Research Team, Centre for Neuroscience, Medical School, University of Pecs, Hungary
2Department of Sportbiology, Faculty of Sciences, University of Pecs, Hungary
Abstract:
Introduction: Type 2 diabetic retinal disease (T2DRD) is a leading cause of permanent blindness in the diabetic population. Pituitary adenylate cyclase-activating polypeptide (PACAP) has neuroprotective effects. One of the isoforms, PACAP38 has an impact against the apoptotic signaling pathways and creates an anti-inflammatory environment in the retina. Our aim was to test the possible retinoprotective effect of topical administration of PACAP38 in type 2 diabetic animal model.
Methods: Type 2 diabetes was induced in Wistar rats with the combination of streptozotocin administration and high-fat diet. All rats were treated topically two times a day for 4 months, accordingly. Diabetes model was validated by fasting oral glucose tolerance test and C-peptide ELISA test. Electroretinography (ERG), optical coherence tomography (OCT), immunohistochemistry, and vessel analysis were performed in the retinal samples.
Results: Our ERG results showed that the visual function of PACAP38-treated diabetes group was preserved. The OCT measurements correlated with the ERG data where the total retinal thickness was preserved in the diabetes+PACAP38 group. The retinal microvascular structure and the ganglion cell number were also protected.
Conclusions: Topically administered PACAP38 has displayed its potent neuroprotective effect against T2DRD, therefore it could be a promising therapeutic approach for the treatment of the disease.
Funding: The research was funded by FK129190, K135457; National Brain Research Program NAP2017-1.2.1-NKP-2017-00002; MTA-TKI-14016; PTE AOK-TANDEM; GINOP-2.3.2-15-2016-00050 “PEPSYS”; EFOP-3.6.2-16-2017-00008; “The role of neuroinflammation in neurodegeneration: from molecules to clinics''; and Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary: 20765/3/2018/FEKUTSTRAT, 2020-4.1.1-TKP2020—FIKP III. Project No. TKP2020-IKA-08 has been implemented with the support provided from the National Research, Development and Innovation Fund of Hungary, financed under the 2020-4.1.1-TKP2020 funding scheme.
Balazs Meresz1, Inez Bosnyak1, Dorottya Molitor1, Bela Kocsis2, Kornelia Farkas3, Dora Reglodi1, Tamas Atlasz1,4, Alexandra Vaczy1
1MTA-PTE PACAP Res. Group, Dept. of Anatomy, University of Pecs Medical School, Pecs, Hungary
2Dept. of Medical Microbiology and Immunology, University of Pecs Medical School, Pecs, Hungary
3Institute of Bioanalysis, University of Pecs Medical School, Pecs, Hungary
4Faculty of Sciences, Dept. of Sportbiology, University of Pecs, Pecs, Hungary
Abstract:
Introduction: Bacterial keratitis is an infection of the corneal tissue caused by various bacterial species. Keratitis is still responsible for the most sight-threatening lesion. Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide, known for its neuroprotective and anti-inflammatory effects, which are mainly mediated through PAC1 receptor (PAC1R). The aim of our research is to investigate the protective role of the PAC1R in endotoxin-induced keratitis model in mice.
Methods: In our study, we induced bacterial keratitis in a CD1-IGS mouse strain by intraperitoneal injection of lipopolysacharide (LPS). To investigate the role of PAC1R, half of the animals were treated intravitreally with a PAC1R antagonist, maxadilan. 24 hours after injection, optical coherence tomography (OCT) and routine histology were performed. Cytokine arrays were executed to map inflammatory pathways. Five weeks after the LPS administration, a four-step scoring system was used to determine the degree of keratitis.
Results: Based on OCT and histology results, maxadilan attenuated the increase of central corneal thickness and inhibited the activation of inflammatory cytokines. Five weeks later the group receiving LPS and treated with maxadilan showed less severe keratitis.
Conclusion: Our results suggest that targeting the PAC1R could be a promising therapeutic approach for the treatment of bacterial keratitis.
Funding: The research was funded by: FK129190, K135457; National Brain Research Program NAP2017-1.2.1-NKP-2017-00002; MTA-TKI-14016; PTE AOK-TANDEM; GINOP-2.3.2-15-2016-00050 “PEPSYS”; EFOP-3.6.2-16-2017-00008; “The role of neuroinflammation in neurodegeneration: from molecules to clinics''; and Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary: 20765/3/2018/FEKUTSTRAT, 2020-4.1.1-TKP2020—FIKP III. Project No. TKP2020-IKA-08 has been implemented with the support provided from the National Research, Development and Innovation Fund of Hungary, financed under the 2020-4.1.1-TKP2020 funding scheme.
Dominika Szlachcikowska, Anna Tabęcka-Łonczyńska
Department of Biotechnology and Cell Biology, Medical College, University of Information Technology and Management in Rzeszów, Rzeszów, Poland
Abstract:
Tris(2,3-dibromopropyl)isocyanurate (TBC) is one of the novel brominated flame retardants (NBFR), widely used in industry. Current data showed significant biaccumulation of TBC in the environment and living organisms. Because it has been showed that TBC is a lipophilic substance, while nervous tissue is rich in fat, so probably TBC can pass through the blood-brain- barrier (BBB). To date, TBC is also known as an endocrine disruptor which may have a particularly negative effect on the nervous system of the human and animals. This may be important for the induction of inflammatory processes that cause disorders and development of neurodegenerative diseases. Therefore, the aim of our research was to evaluate the effect of TBC treatment on the mouse hippocampal neuronal cell line (HT-22) in vitro as a model cells. It is also important to understand the mechanism of action of TBC. The basic parameters of the research were changed in HT-22 cells after use TBC with cotreatment. Additionally, we analyzed the effect of TBC and cotreatment on the expression of proteins related to inflammatory processes. Our results indicate the involvement of TBC in these processes, which may indicate a negative impact on neuronal cells.
Funding: This work was supported by statutory funds from the University of Information Technology and Management in Rzeszow, Poland (DS 503-07-01-59).
Katarina Bobić1, Dunja Drakulić1, Snežana Pejić1, Tijana Milovanović2, Jadranka Miletić Vukajlović3, Snežana Pajović1, Ana Todorović1
1Department of Molecular Biology and Endocrinology, “VINCA” Institute of Nuclear Sciences - National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Republic of Serbia
2Public Company “Nuclear Facilities of Serbia”, Belgrade, Republic of Serbia
3Department for Physical Chemistry, “VINCA” Institute of Nuclear Sciences - National Institute of the Republic of Serbia, University of Belgrade, Belgrade, Republic of Serbia
Abstract:
The broad use of titanium(IV) oxide (TiO2) particles in various products, and toxicity indicators of particles with diameters < 100 nm (nanoparticles, NPs), inspire scientists to find a suitable, non-toxic replacement. An effective approach to diminish TiO2 NPs toxic impact might comprise altering their physicochemical properties by surface modification. Therefore, commercially available TiO2 NPs were surface-modified with antioxidants: salicylic acid (SA-TiO2) or 5-aminosalicylic acid (ASA-TiO2), with a goal to compare their ability to reduce oxidative stress, specifically levels of lipid peroxidation (LPO) and advance oxidation protein products (AOPP) in the brain of Wistar rats. Oxidative stress markers were determined in whole cell brain extracts, isolated 14 days following acute oral intake of either vehicle (V, 2.5 ml 0.01 M HCl) or TiO2/SA-TiO2/ASA-TiO2 NPs (1000 mg/kg dissolved in V). The results showed that LPO levels were evenly elevated in all TiO2 treated groups relative to the V group. However, AOPP levels in ASA-TiO2 group were similar to V group and significantly lower compared to TiO2 and SA-TiO2 groups; suggesting that this modification moderates some of the TiO2 NPs-induced toxicity. Additional experiments are required to investigate their effects on toxicity mechanisms and fully elucidate the potential of ASA-TiO2 as TiO2 NPs substitute.
Funding: Ministry of Science, Technological Development and Innovation of the Republic of Serbia, grant 451-03-66/2024-03/200017.
Bartosz Skóra, Karolina Kosińska, Konrad A. Szychowski
Department of Biotechnology and Cell Biology, Medical College, University of Information Technology and Management in Rzeszow, Poland
Abstract:
Elastin is present in many tissues e.g. brain. It is well-described that elastin-derived peptides (EDPs) are created during age due to elastin decay. The increasing amount of EDPs during aging correlates with the first symptoms of many neurodegenerative diseases. Therefore, the study aimed to determine the impact of EDP on basic parameters of neuroblastoma-derived neurons, together with determining its ability to induce neurodegeneration in these cells as well as engagement of certain roles of the specific histone deacetylase (HDAC) in this process. The obtained results show no significant changes in the cells during dose-response analysis after 24h and 48h of treatment, however, the confocal microscope-based analysis proved the ability of the EDP to induce morphological changes in the neurons i.e. shortening axons' length. Furthermore, the changes in the antioxidant enzymes were detected in the tested time intervals of treatment. Lastly, the increased expression of HDAC and tubulin-related proteins were observed. Summarizing, the results show - for the first time - that tested EDP can cause neurodegeneration phenotype in neurons with potential engagement of specific HDAC. However, more studies are needed in this field.
Funding: Statuary funds of University of Information Technology and Managament in Rzeszow, DS.: 503-07-01-77.
Anjani Kumar Tiwari1.2,3, Anil Mishra2, Ming R. Zhang3
1Department of Chemistry, Babasaheb Bhimarao Ambedkar University, Lucknow, Uttar Pradesh, India
2Division of Cyclotron and Radiopharmaceutical Sciences (DCRS), Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, New Delhi, India
3Department of Advanced Nuclear Medicine Sciences, Institute for Quantum Medical Science, National Institutes for Quantum Science and Technology, Japan
Abstract:
The 18kDa Translocator protein (TSPO), formerly recognized as the peripheral benzodiazepine receptor (PBR), has been considered a positron emission tomography (PET) biomarker for various types of neuroinflammation. While it has been demonstrated that TSPO ligands may decrease the level of beta-amyloid peptide associate in Alzheimer's but their role for tau pathology remains unexplored. To investigate this aspect, we conducted an analysis of the influence of the third-generation TSPO ligand, BBMP regarding the progression of the tau transgenic mice neuropathology.
Methods: In pursuit of this objective, we conducted an analysis of the impact of the third-generation TSPO ligand BBMP (2-(5-Bromo-2-oxo-1,3-benzoxazol-3(2H)-yl)-N-methylN-phenylacetamide) on development of neuropathology in TauTg. Analysis were made through MR and PET imaging and immunohistochemistry/autoradiography.
The in vivo animal neuro-imaging results were corroborated with immunohistochemistry to analyse markers associated with inflammation (TSPO, Controller of the complement cascade (1q / C1q), microglial tauopathy marker (AT8) and the neuronal survival (NeuN), in the brain sections obtained from the mice that underwent scanning.The administration of BBMP significantly reduced atrophy during the 6 to 10-month period, resulting in a 50% reduction in cortical and hippocampal volumes in comparison to TauTg mice administered with the vehicle
Administering BBMP to TauTg mice resulted in a decrease in average concentration levels of the three assessed inflammatory markers. Distance-based redundancy analysis revealed a strong correlation (80%) between the variation in inflammatory markers among groups and C1q (0.996). The indicators of neuroinflammation/degeneration in cortex and hippocampus exhibited a negative association with inflammatory markers.
In conclusion, our data point to a safeguarding role of new generation TSPO ligands in tauopathies, with the potential to lower neuroinflammation/degeneration, and brain atrophy.
Naira Rashid and Sayeed ur Rehman A Kalam
Department of Biochemistry, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, India
Abstract:
HSPB8 also known as the Alpha-crystallin C chain , Protein kinase H11 or Small stress protein-like protein HSP22 is a small heat shock protein. This protein functions as a chaperone in association with Bag3, a stimulator of macroautophagy. It intervenes in the governance of cell proliferation, apoptosis, and carcinogenesis. Mutations in this gene have been linked with various neurodegeneration. HSPB8 manoeuvres its role by expediting autophagy by inhibiting the deposition of misfolded proteins in afflicted cells. Ameliorating the discerning attempt of misfolded proteins by HspB8-BAG3-Hsp70 to autophagy might also result in reduced delivery to the proteasome by the BAG1-Hsp70 complex so that the probable proteasome overwhelming is reduced.
With the help of bioinformatics, we have predicted novel exons of the human HSPB8 gene. The addition of novel exons in the mature RNA transcript resulted in the removal of the essential residues of the HSPB8 protein. These novel isoforms can be analysed to understand their role in neuroprotection and several other processes in which HSPB8 plays a cardinal role. These novel isoforms can also be targeted for therapeutic purposes by developing splice-switching antisense oligonucleotides (SSOs) in order to regulate their functioning.
Michal Rafal Zareba1, Krzysztof Bielski2, Victor Costumero1, Maya Visser1
1Department of Basic and Clinical Psychology and Psychobiology, Jaume I University, Castellon de la Plana, Spain
2Institute of Psychology, Jagiellonian University, Krakow, Poland
Abstract:
Maladaptive forms of guilt, such as excessive self-blame, are common characteristics of anxiety disorders. Nevertheless, despite understanding the anatomy of the guilt processing circuitry, it is currently unknown how its network-level interactions are related to subclinical anxiety and self-blaming behaviour. To fill this gap, we used resting-state functional and diffusion-weighted magnetic resonance imaging (MRI) data to construct the guilt networks in 78 young healthy adults, and subsequently investigated associations of their graph theory parameters with these two phenomena. Increased functional contributions (higher clustering coefficient, local efficiency and strength) of the left superior anterior temporal lobe (sATL) were a common feature for individuals with higher self-blaming and trait-anxiety, while functional isolation (lower clustering coefficient and local efficiency) of the left inferior frontal gyrus pars opercularis and insula was related to higher trait-anxiety. Trait-anxiety was also linked to the structural network’s global parameters (mean clustering coefficient), with the circuitry’s architecture favouring increased local information processing in individuals with increased anxiety levels. Previous research suggests aberrant interactions between conceptual (sATL) and affective (fronto-limbic) regions underlie maladaptive guilt and the current results align and expand on this theory by detailing network changes associated with self-blame and trait-anxiety.
Amelia Jasińska¹, Łukasz Krzywoszański², Radosława Herzog-Krzywoszańska³
¹Uniwersytet Komisji Edukacji Narodowej, Kraków, Poland
²Institute of Psychology, Uniwersytet Komisji Edukacji Narodowej, Kraków, Poland
³Institute of Psychology, Uniwersytet Komisji Edukacji Narodowej, Kraków, Poland
Abstract:
Current knowledge of Gray’s Reinforcement Sensitivity Theory (RST) and stimulating substances is mainly focused on drugs and alcohol. Relationships between BIS/BAS and caffeine have not yet been researched. The study explored relationships between RST, caffeine consumption, and self-control in 187 Polish adults aged 18 to 30 years old. A Polish version of the RSQ (Reinforcement Sensitivity Questionnaire), a Polish version of the SCS (Self-Control Scale) and a self-made caffeine intake questionnaire were used. Questionnaires were sent out online in the form of a survey.
Relationships between caffeine and BIS/BAS scores were not determined. No relationships between caffeine intake and self-control were found.
There were no differences between subjects with high and low scores on the sensitivity to reward (BAS) scale in caffeine intake.
There was a moderate negative correlation between the sensitivity to punishment (BIS) scale and self-control. Additionally, there were significant differences between people with high and low BIS scores; people with low BIS had higher scores on a self-control scale than people with high BIS. Individuals with high BIS might be driven by the avoidance of risk and punishment, which takes away control of their own actions.
The study highlights the lack of research between RST and caffeine intake.
Mehlika Merve Yolcu
Psychological Counselling and Guidance, Institute of Graduate Education, Istanbul Sabahattin Zaim University, Istanbul, Turkey
Psychology, Emotion Regulation and Parenting Lab, Norwegian University of Science and Technology, Trondheim, Norway
Abstract:
This study, aims to examine the relationship between the level of intolerance for uncertainty and the affective neuroscience personality traits. The sample is a total of 174 participants, aged between 18-25, female (112) and male (62) in Turkey. It was used a non-experimental cross-sectional and correlational design in this study. Demographic data were collected using a Demographic Information Form, Affective Neuroscience Personality Scale and Intolerance of Uncertainty Scale were used to data collection. Positive correlations were identified between Intolerance of Uncertainty and emotions such as SAD, ANGER, FEAR and CARE, elucidating the emotional associations of uncertainty in the affective neuroscience personality scale. It is planned that this study be expanded into a cross-cultural investigation by collecting data from Norway in the future. This proposed research aims to explore tolerance to uncertainty within the context of cross-cultural affective neuroscience, focusing on the influence of culture on the regulation of basic emotional systems. By explaining how tolerance for uncertainity is influenced by cultural context and how it relates to emotions in affective neuroscience, intends to contributes to improving cross-cultural understanding and insights into its implications for mental health outcomes like anxiety disorders, guiding interventions to enhance resilience and coping mechanisms.
Beyza Nur Salcıoğlu, Malwina Ankiewicz and Halszka Ogińska
Department of Cognitive Neuroscience and Neuroergonomics, Institute of Applied Psychology
Jagiellonian University, Krakow, Poland
Abstract:
Sleep deficit results in a variety of neurocognitive effects, impaired emotional recognition and psychosocial functioning. This study explores individual differences in vulnerability to sleep deprivation in terms of subjective socio-emotional and cognitive functioning.
Thirty healthy volunteers (mean age 23.4 ± 3.7 years) participated in three sleep conditions: full-rest sleep/baseline (B), chronic partial sleep deprivation (C), and acute/total sleep deprivation (A). Two sets of subjective questionnaires assessed individual traits (chronotype, sensitivity, and personality dimensions) and well-being measures including sleepiness, physiological symptoms, irritation, cognitive attenuation, positive and negative affectivity, and pleasure experience.
Results indicated varying physiological symptoms, irritation, and cognitive attenuation across conditions, with total sleep deprivation showing the largest effects. Differences in pleasure were observed between full-rest and chronic sleep deprivation conditions, participants reported less pleasure in the latter.
As far as subjective well-being is concerned, total sleep deprivation seems to be a more difficult experience than chronic sleep restriction. Resistance to chronic sleep restriction is linked to morning orientation, low sleep need, low sensitivity, and high extraversion. Resistance to total sleep deprivation is associated with low circadian rhythm amplitude and lower levels of sensitivity, neuroticism, agreeableness, and openness to experience. These findings contribute to a better understanding of the individual response to sleep loss.
Funding: The research was supported by the grant from the Polish National Science Centre No 2018/29/B/HS6/01934.
Patrycja Ściślewska1,2, Michał Rafał Zaręba3, Julia Lengier1, Aaron Schirmer4, Piotr Bębas1, Iwona Szatkowska2
1Department of Animals Physiology, Faculty of Biology, University of Warsaw, Warsaw, Poland
2Laboratory of Emotions Neurobiology, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland
3Department of Basic and Clinical Psychology and Psychobiology, Jaume I University, Castellon de la Plana, Spain
4Northeastern Illinois University, Dept. of Biology, Chicago, USA
Abstract:
Several studies found behavioral and neural differences corresponding to chronotypes. However, the impact of other dimensions of the circadian rhythm is unclear. This study explored how circadian rhythm distinctness (subjective amplitude) influenced reward and punishment processing. We examined 37 healthy participants aged 20-30. Distinctness was measured using the Morningness-Eveningness Stability-Scale improved (MESSi) questionnaire. We also administered questionnaires assessing personality (NEO-Five Factor Inventory; NEO-FFI) and reinforcement processing: the Sensitivity to Punishment and Sensitivity to Reward Questionnaire (SPSRQ), and the Positive and Negative Affect Schedule (PANAS). Moreover, we used functional magnetic resonance imaging (fMRI) during the Monetary Incentive Delay Task, which is a common method of assessing reward- and punishment-motivated behavior. Analyses were conducted using the FSL program and statistical packages in Python. We found a positive correlation between a value of distinctness and a level of neuroticism, sensitivity to punishment, and negative affect. fMRI results showed increased activity during the anticipation phase of ‘punishment’ trials in the bilateral Superior Frontal Gyrus and Supplementary Motor Cortex (BA 8, 6) for higher values of the distinctness. The results emphasize the negative emotionality aspect of high-distinctness people and suggest the association between a higher value of distinctness and enhanced neural processing of punishment-related information.
Funding: This work was supported by the Ministry of Science and Higher Education (Poland) as a project under the program Excellence Initiative – Research University (2020-2026) no. BOB-IDUB-622-28/2023 (IV.4.1.).
Chris Townsend, Stephane De Brito
Centre for Human Brain Health, School of Psychology, University of Birmingham,
Birmingham, UK
Abstract:
Psychopathy is a personality disorder that manifests as a syndrome characterized by a constellation of affective, interpersonal, and behavioral features. Every conceptualization of psychopathy considers impulsivity a central feature of the disorder. However, both psychopathy and impulsivity are multifaceted constructs and associations between the different facets of those constructs has never been subjected to a systematical assessment. We conducted a pre-registered meta-analysis to estimate the direction and the strength of association between psychopathy and impulsivity. Moderator analyses included factors of psychopathy, measures of impulsivity, demographic characteristics of the participants, recruitment sources, as well as measures of the disorder. One-hundred and sixty-eight articles, consisting of 173 independent samples (N = 43,944) were included. The overall association between psychopathy and impulsivity was reliable and yielded a moderate effect size (r = .23), with a stronger association for Trait (r = .27) than State (r = .16) measures with no evidence of publication bias. Factor 1 had a weaker association (r =.10; no Trait-State moderation) than Factor 2 (r = .26; Trait-State moderation: Trait: r = .30; State: r = .13). We show that impulsivity is positively associated with psychopathy, but this association varies by psychopathy factors and the type of impulsivity measure.
Daria Nowak1, Natalia Kruszyna1, Jakub Nastaj1, Jacek Skalski1, Tibor Szikszay2, Kerstin
Luedtke1,2, Wacław M. Adamczyk1
1Laboratory of Pain Research, Institute of Physiotherapy and Health Science, The Jerzy Kukuczka Academy of Physical Education, Katowice, Poland
2Institute of Health Sciences, Department of Physiotherapy, Pain & Exercise Research Lübeck (P.E.R.L), University of Lübeck, Lübeck, Germany
Abstract:
One of the most challenging health conditions to address is chronic widespread pain, the mechanisms of which are not fully understood. We used classical conditioning procedures to investigate if the expanded area of pain results from classical conditioning between nociceptive stimuli originating from different nociceptive foci.
Healthy adult participants were randomly assigned to one of four experimental groups. In the first group, participants underwent classical conditioning by pairing visual cues with the magnitude of the painful area. They were also informed, through verbal instruction, that one of the visual cues would announce pain distributed at a larger area. In the second group, participants were exposed only to classical conditioning, while in the third group, they only received verbal suggestion. The fourth group served as a control (no conditioning, no instruction). Nociceptive stimuli were delivered through electrodes attached to participants’ hands, activating two electrodes for a large distribution or one for a small distribution of pain.
Classical conditioning combined with instruction was effective at inducing pain at a larger area despite nociceptive stimulation targeting a small area of the body (p < 0.001). Furthermore, this effect was preserved even if two methods were used separately: Classical conditioning alone (p < 0.05) or verbal suggestion alone (p < 0.01) produced conditioned pain distribution. We did not find a statistical effect in the control group (p > 0.05).
Funding: The study was funded by a OPUS19 grant awarded by the National Science Centre in Poland (no. 2020/37/B/HS6/04196).
Wiktoria Rabińska1, Dennis Larsson2, Henrik Ehrsson2
1Institute of Psychology, Jagiellonian University, Kraków, Poland
2Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden
Abstract:
The full-body illusion is a well-established experimental paradigm for inducing illusory ownership over fake or virtual bodies. While subjective ratings and proprioceptive drift have been used to measure this phenomenon, our study aimed to explore the potential of the cross-modal congruency effect (CCE) as an alternative measurement of body ownership. In our study, participants wore VR goggles displaying a mannequin's body and felt vibrations on their feet. Red dots appeared on the mannequin’s feet simultaneously, congruently or incongruently with the felt vibration, and participants’ task was to report immediately which foot they felt the vibration on. Additionally, they received synchronous or delayed stroking on their abdomen and thighs, while observing the same actions in VR. While synchronous trials yielded stronger subjective rating of ownership in synchronous trials and faster CCE task response time during congruent trials, there was no significant main effect of illusion condition or interaction between illusion condition and congruency on CCE task reaction time. These findings suggest successful induction of the illusion through the first-person perspective, yet the lack of correlation with CCE implies a dissociation between body ownership and cross-modal congruency. Further research is needed to explore the relationship between CCE, peripersonal space, and self-location experiences.
Department of Neurobiology and Biophysics, Vilnius University, Vilnius, Lithuania
Abstract:
The dual-hormone hypothesis states that testosterone is positively related to status-seeking behaviors only when cortisol level is low. This idea has been validated in the research on status-relevant behaviors. However, there are no studies investigating this hypothesis in the context of social stress perception. Socially stressful situations might be perceived as a status threat. Therefore, joint effects of testosterone and cortisol are plausible. This study aimed to evaluate if the dual-hormone hypothesis applies to the subjective evaluation of perceived social stress.
Mild social stress was induced in 36 males by using a short Sing-a-Song Stress Test. An electrocardiogram was recorded to evaluate heart rate variability (HRV) parameters. Participants indicated perceived stress on the visual analog scale. Before the stress manipulation, saliva samples were collected to measure basal levels of testosterone and cortisol.
HRV analysis confirmed stress induction. Moderation analysis revealed a negative association between testosterone and perceived stress when cortisol levels were low. Whereas when cortisol levels were high, testosterone was positively associated with perceived stress. Cortisol was negatively related to perceived stress only when testosterone levels were low. These findings support the dual-hormone hypothesis on the perception of social stress.
1Department of Behavioural Neuroscience, Centre of Experimental Medicine, Slovak Academy of Sciences, Bratislava, Slovakia
2Department of Psychiatry, Faculty of Medicine, Slovak Medical University, Bratislava, Slovakia
Abstract:
To examine how the fear of falling influences balance control independently of aging and pathology, healthy individuals are exposed to various postural threats, representing natural stressors. The most common postural threat is exposure to height by elevating the surface on which individuals stand. The aim of the study was to examine postural and psychophysiological responses to virtual height exposure in 42 young individuals with different intensities of fear of heights. The measurements were carried out during stance on an unstable surface at ground level and at heights of 20 m and 40 m in a virtual reality environment. The height exposure elicited a complex, robust, and reliable psychophysiological response with significant changes in emotional state, sympathetic activity, and postural control, which were enhanced in individuals with a fear of heights. Those with higher fear showed a rigid and more stiffened posture characterized by smaller magnitude and simultaneously increased velocity of body sway. For the assessment of postural stability in stressful environments, such as height exposure (both real and virtual), it is important to take into account the currently experienced fear and anxiety that can significantly contribute to resulting postural stability and may have more significant implications in populations with a fear of falling.
Funding: This work was supported by the grants VEGA 2/0080/22 and APVV-20-0420.
Paulina Lewandowska1,8, Claude J. Bajada2, Mikołaj Compa1, Aleksandra Domagalik3, Katarzyna Kaczmarek-Majer4, Krzysztof Skotak4, Małgorzata Lipowska5,6, Clemens Baumbach1, Yarema Mysak1, Katarzyna Sitnik-Warchulska6, James Grellier1,7, Iana Markevych1,9,10 and Marcin Szwed1
1Institute of Psychology, Jagiellonian University, Krakow, Poland
2Department of Physiology and Biochemistry, Faculty of Medicine and Surgery, University of Malta
3Brain Imaging Core Facility, Małopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland
4Institute of Environmental Protection-National Research Institute, Warsaw, Poland
5University of Gdansk, Gdansk, Poland
6Faculty of Management and Social Communication, Institute of Applied Psychology, Jagiellonian University, Krakow, Poland
7European Centre of Environment and Human Health, University of Exeter Medical School, Royal Cornwall Hospital, Truro, UK
8Doctoral School of Social Sciences, Jagiellonian University, Krakow Poland
9Environmental Health Division, Research Institute at Medical University of Plovdiv, Medical University of Plovdiv, Plovdiv, Bulgaria
10Environmental Health Division, Research Institute at Medical University of Plovdiv, Medical University of Plovdiv, Plovdiv, Bulgaria
Abstract:
Air pollution has been proven to have a negative impact on human health. There is evidence that it can cause cardiovascular disease, pulmonary disease, and breathing problems but there is still little known about how air pollution affects the nervous system.
Here, we present preliminary results on how early life (0-4 years) and prenatal air pollution exposure affects global brain measures - fractional anisotropy and mean diffusivity - in children aged 10 to 13 from the NeuroSmog project. We found significant associations between early life NO2 exposure and global white matter integrity measures: fractional anisotropy, and mean diffusivity. There were no significant associations between prenatal exposure to NO2 or early life and prenatal exposure to PM10 and any of the global measures. Our result is in line one previous study and suggests that air pollution exposure during the crucial developmental period may have a negative effect on the development of the children’s brain. It is a prelude for the main fixel-specific whole brain analysis.
Funding: Supported by “NeuroSmog: Determining the impact of air pollution on the developing brain” (Nr. POIR.04.04.00-00-1763/18) grant to Marcin Szwed implemented as part of the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources obtained from the European Regional Development Fund under the Smart Growth Operational Programme.
Mikołaj Compa1,8, Aleksandra Domagalik3, Yarema Mysak1, Katarzyna Kaczmarek-Majer4, Krzysztof Skotak4, Małgorzata Lipowska5,6, Clemens Baumbach1, Katarzyna Sitnik-Warchulska6, James Grellier1,7, Iana Markevych1,9,10 and Marcin Szwed1
1Institute of Psychology, Jagiellonian University, Krakow, Poland
2Department of Physiology and Biochemistry, Faculty of Medicine and Surgery, University of Malta
3Brain Imaging Core Facility, Małopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland
4Institute of Environmental Protection-National Research Institute, Warsaw, Poland
5University of Gdansk, Gdansk, Poland
6Faculty of Management and Social Communication, Institute of Applied Psychology, Jagiellonian University, Krakow, Poland
7European Centre of Environment and Human Health, University of Exeter Medical School, Royal Cornwall Hospital, Truro, UK
8Doctoral School of Social Sciences, Jagiellonian University, Krakow Poland
9Research Group “Health and Quality of Life in a Green and Sustainable Environment”, Strategic Research and Innovation Program for the Development of MU – Plovdiv, Medical University of Plovdiv, Plovdiv, Bulgaria
10Environmental Health Division, Research Institute at Medical University of Plovdiv, Medical University of Plovdiv, Plovdiv, Bulgaria
Abstract:
Air pollution is one of the main environmental hazards to human health. Toxic pollutants in the air not only affect lungs and cardiovascular systems, but can also negatively impact attention, and brain structure. In the present study we investigated whether long-live and early life exposure to air pollution - nitrous dioxide (NO2) and particulate matter (PM10) - has impact on functional brain activity during an inhibition control task in 447 10–13-year-old children with and without diagnosis of ADHD from the NeuroSmog study. We found that exposure to PM10, but not NO2 was related to decreased task-related activity in right anterior cingulate cortex and right pre-supplementary motor area, regions typically related to attention. To our knowledge, this is the first demonstration of a fMRI task-related activity deficit related to air pollution exposure.
Funding: This work was supported by the “NeuroSmog: Determining the impact of air pollution on the developing brain” (Nr. POIR.04.04.00-1763/18-00) grant to Marcin Szwed implemented as part of the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources obtained from the European Regional Development Fund under the Smart Growth Operational Programme. Additional funding came from a grant from the Priority Research Area (“Anthropocene”) to Marcin Szwed under the Strategic Programme Excellence Initiative at Jagiellonian University, Poland.
Sylwia Adamus1,2,3, Małgorzata Draps3, Krzysztof Bielski4, Mateusz Gola3,5
1Faculty of Medicine, Medical University of Warsaw, Warsaw, Poland
2Faculty of Physics, University of Warsaw, Warsaw, Poland
3Institute of Psychology, Polish Academy of Sciences, Warsaw, Poland
4NeuroEconomics Research Group, Institute of Psychology, Faculty of Philosophy, Jagiellonian University, Cracow, Poland
5Swartz Center for Computational Neuroscience, Institute for Neural Computations, University of California San Diego, San Diego, CA, USA
Abstract:
Compulsive Sexual Behavior Disorder (CSBD) is an impulse control disorder characterized by the inability to resist engaging in various sexual behaviors. It has been suggested that the amygdala could play an important role in the pathogenesis of this disorder.
This study aimed to assess whether resting-state functional connectivity of the amygdala subdivisions’ is correlated to the severity of CSBD symptoms.
An amygdala parcellation based on Recurrence Quantification Analysis was performed on resting-state fMRI data from 45 heterosexual CSBD male patients. The severity of CSBD was assessed by the Sexual Addiction Screening Test-Revised (SAST-R) and its score was used as a covariate in seed-to-voxel connectivity analysis with amygdala subdivisions as regions of interest.
The SAST-R score negatively correlated with the resting-state functional connectivity between the Right Dorsomedial Amygdala and Left Middle & Inferior Temporal Gyri and Right Angular Gyrus. Such a negative correlation was also observed between the Left Ventrolateral Amygdala and Left Postcentral Gyrus.
To the best of our knowledge, this is the first attempt to investigate the amygdala of CSBD patients via a parcellation pipeline, which provides a new insight into neural mechanisms underlying this disorder.
Funding: Data collection was supported by the Polish National Science Centre OPUS grant (2014/15/B/HS6/03792) to MG, and MD is supported by the Foundation for Polish Science (FNP), scholarship number START 014.2023.
Agata Szymańska, Beata Pacula-Leśniak
Emotion and Perception Lab, Institute of Psychology, Department of Philosophy, Jagiellonian University, Krakow, Poland
Abstract:
In today's digital age, authenticity assessment of media content faces challenges due to the blend of traditional and social media, and the rise of deep fakes.
we aimed to uncover cognitive processes engaged in evaluating the authenticity of the photo in a media-centric environment by investigating the impact of the emotional nature of the photograph on its authenticity assessments and examining the correlation with political orientation of participants, We have shown 1000 contemporary press photographs - all of them real - to 300 people from the general population and asked them to classify each as real or fake. In a subsequent online study 600 participants rated those photos on valence and arousal scales, as well as marked emotions they felt while viewing the picture. At the end participants filled the political orientation questionnaire with economic and cultural subscale.
Anger, compassion, disgust, excitement, fear, guilt, hope, pain, sadness, and tenderness significantly correlated with image authenticity score. Women reported more sadness, fear, and regret compared to men, while men reported more excitement and disgust. Depending on polarization on the cultural and economic subscale, individuals also reported feeling different emotions. Nonetheless, our results highlight how emotions, gender, and political beliefs may differently influence media perception.
Funding: NCN Grant Project OPUS edition 39, 2020/39/B/HS6/03324
Izabela A. Łaska¹, Aleksandra Domagalik-Pittner², Halszka Ogińska³
¹Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Cracow, Poland
²Centre for Brain Research, Jagiellonian University, Cracow, Poland
³Department of Cognitive Neuroscience and Neuroergonomics, Institute of Applied Psychology, Jagiellonian University, Cracow, Poland
Abstract:
Food intake is regulated by two complementary drives: the homeostatic pathway based on energy balance regulation, and hedonic involving the brain reward system. Hedonic regulation can override the homeostatic pathway during energy abundance by increasing the desire to consume foods that are highly palatable. Sleep is bi-directionally linked to energy balance. The mechanisms mediating the relationship between sleep duration and food intake are likely multifactorial and include differences in appetite-related hormones, hedonic pathways, extended hours of food intake. The present study examined the effects of chronic sleep deprivation on appetite regulation by analyzing subjective ratings of visual stimuli presenting food. Thirty healthy volunteers (mean age, 23.4 ± 3.58 years; 57% females) took part in a study that compared their responses in two conditions: chronic sleep deficit (5 nights with sleep restricted to 5 hours) and regular sleep. In both conditions, participants were presented with 64 visual stimuli representing various high-energy and low-energy foods.
Based on our findings, we can say that subjective appetizing ratings of foods increase with the levels of subjective sleepiness and not with objective sleep curtailment.
It can also be observed that appetite ratings do not correlate with the actual level of hunger - these represent separate mechanisms.
Funding: This research was supported by the grant from the Polish National Science Centre Nr 2018/29/B/ HS6/01934.
Martyna Piasecka
School of Psychology, Institute of Neuroscience, Newcastle University, Newcastle Upon Tyne, UK
Abstract:
Approximately 3 million people in the UK suffer from the most common red/green colour-vision
deficiency (CVD). It affects different aspects of lives, including education, social interactions, and general quality of life. Children with CVD may perform poorly on colour-related tasks in comparison with their peers. This study aimed to investigate differences in self-esteem between school children with CVD and with normal colour vision. Children were tested using battery of colour vision tests (Ishihara Plates, Anomaloscope, Farnsworth-Munsell D15, Medmont C-100). Their self-esteem was measured with MMG, which includes two motives: achievement (Hope for Success (HS), Fear of Failure (FF)) and affiliation (Hope for Affiliation (HA), Fear of Rejection (FR)). Data from 35 CVD participants and 60 controls was analysed. An independent 2-samples t-test was performed to test for significant differences in their MMG scores. Results of the main analysis did not show a significant difference. Additional analyses revealed a significant difference between CVD students and controls in the FF motive in art scenario (p<.05). The main limitation of this study was the small sample size. Further research should consider different types of CVD and its severity independently and account for age.
Michał Remiszewski, Gabriela Rajtar, Tomasz S. Ligęza
Institute of Psychology, Jagiellonian University, Cracow, Poland
Abstract:
Physical exercise offers many benefits for both physical and mental health. Recent studies have demonstrated that acute exercise positively affects emotional responsiveness by enhancing neural responses to positive stimuli while decreasing responses to negative stimuli. However, it remains an open question whether long-term exercise exerts a similar influence on emotional responsiveness. To address this gap, we conducted a longitudinal experiment in which 53 participants either engaged in a series of aerobic exercises over a 1.5-month testing period (18 training sessions in total) (n = 23, experimental group) or maintained their usual habits (n = 30, control group). Before and after intervention participants underwent a passive picture presentation task involving viewing positive, neutral, and negative images. EEG was concurrently recorded to assess a neural marker of emotional responsiveness: LPP component. Findings revealed a significant effect of exercise in one of the subcategories of pictures. Specifically, participants in the experimental group exhibited a lower amplitude of the LPP component in the posttest compared to the pretest in response to erotic images. We speculate that these findings may suggest a decreased sexual drive, possibly resulting from the energy expended during exercise. The evolutionary explanation of the results may be a potential energy-saving mechanism.
Funding: Project funded by NCN: Heart & brain interactions as a new approach to understanding the relationship between exercise and mood, 2021/43/D/HS6/02959.
