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1

Behavior and Metabolism Research Laboratory at Mossakowski Medical Research Centre

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Olszynski K. H., Filipkowski R. H.
Acta Neurobiologiae Experimentalis
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2014
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tom 74
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nr 3
Our laboratory provides a venue to conduct behavioral and metabolism research on mice and rats. We offer all levels of help concerning designing appropriate behavioral/metabolism experiments, training or conducting the experiments as well as the analysis, discussion and presentation of the results. Various forms of cooperation within our institution and with external investigators are possible. Please visit http://www. imdik.pan.pl/pl/pracownie-srodowiskowo-uslugowe/srodowiskowe-laboratorium-behawioralno-metaboliczne for the full list of equipment, behavioral models and description of metabolism unit. The equipment assembled allows us to run sensorimotor, motor & anxiety (open field, elevated-plus maze), learning & memory (e.g., fear conditioning, avoidance, Morris water maze) and social tests. Additionally, Automated Blood Sampling and Telemetry system for full automated, chronic metabolic and telemetric experiments on rodents provides: (i) automated system for blood and other body fluids in vivo sampling and collecting in cooling collector, (ii) automated system of solution infusions to vessels or other selected locations in animal and (iii) telemetry able to measure various physiological parameters (blood pressure, biopotentials, etc.). Behavior and Metabolism Research Laboratory equipment is located in air-conditioned, partly noise-attenuated rooms with regulated day/ night cycle.
2

Fear conditioning affects reaction to ultrasonic signals in SHR and wistar rats

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Wardak A., Polowy R., Grymanowska A., Filipkowski R. K., Olszynski K. H.
Acta Neurobiologiae Experimentalis
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2019
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tom 79
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nr Suppl.1
INTRODUCTION: Ultrasonic vocalizations (USV) are means of communication between rats. We are studying them by presenting USV or artificial tones from a speaker (playback experiments) and observing vocal (rat’s own USV), behavioral, and cardiovascular (heart rate, HR) reactions. We used Wistar rats, which are common in USV experiments, and SHR (spontaneously hypertensive rats), whose USV habits have not been investigated. AIM(S): We are especially interested in the role of the autonomic nervous system. Therefore, we are investigating the impact of fear conditioning which affects autonomic balance in Wistar rats and SHR with higher activity of the sympathetic system. METHOD(S): Three different protocols were used (1x, 6x, or 10x; 1 mA, 1 s shocks) and later, the animals were presented with 50 kHz (appetitive) or 22 kHz (aversive) USV. On the day of conditioning, Wistar rats emitted 22 kHz USV immediately after the first electric impulse, while SHR remained silent typically to the sixth-eight shock. Levels of freezing were similar in both strains. On the following day, during ultrasonic signals presentation, after 50 kHz USV playback, SHR did not show a rise in HR nor an increase in their own USV emission, which were both observed in Wistar rats. Both strains responded to 22 kHz USV by a decrease in HR, independently of fear protocol. During the conditioning test, the day after playback experiments, Wistar rats showed lower HR following 1x conditioning. Also, a dramatic rise in numbers of USV was observed in some of 6x animals. Only the HR of 1x conditioned Wistar rats was lower than in the control (not conditioned) group (HR of 6x and 10x Wistar rats did not differ from control group), while in SHR, all conditioned groups tended to have higher HR than controls. CONCLUSIONS: We confirm that fear conditioning affects the reaction to ultrasonic signals in SHR and Wistar rats. Presumably the autonomic nervous system participates in reactions to USV playback; however, further research with pharmacological agents is essential. To our best knowledge, these are the first studies about USV in SHR. FINANCIAL SUPPORT: This work was supported by National Science Centre, Poland, grant no. 2015/19/B/ NZ4/03393.
3

Ultrasonic and cardiovascular responses to ultrasonic sounds and vocalizations

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Filipkowski R. K., Polowy R., Malz M., Gawrys O., Olszynski K. H.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: Ultrasonic vocalizations (USV) of adult rats are thought to be means of social communication and are divided into two categories, 55 kHz and 22 kHz, signaling, respectively, appetitive and aversive states. These states are also known for changes in the heart rate (HR). The autonomous system has a role in both USV shaping and HR response. A common signaling pathway via the vagus nerve connects the laryngeal muscles and the heart. This causes an overlap in HR parameters and many behavioral reactions (the polivagal theory). AIM(S): The aim of this study was to investigate USV emissions and HR changes in rats evoked by USV presentation. METHOD(S): Ten weeks old Wistar male rats were housed in pairs or separately for 4 weeks. Telemetry transmitters for HR acquisition were implanted in the peritoneum with the detector placed in the aorta. Rats were exposed to five 10‑s sets of sounds (counterbalanced): 55-kHz, 22-kHz USV (both natural, collected from other animals), 55-kHz, 22-kHz tones, 22-kHz uninterrupted tone (all three artificial, software‑generated) separated with 5 min silence intervals. HR and USV emitted were registered. RESULTS: Rats of both groups responded with USV mostly and more often to 55-kHz tones and vocalizations than during presentation of 22-kHz sounds. The responses were, almost exclusively, within 55 kHz range. In general, single-housed rats vocalized more often than pair-housed ones but the effect was not strong. Also, HR changes were more pronounced following presentation of natural USV. During 55-kHz USV presentation, there was an elevation of HR in single-housed animals, while in pair-housed animals, this elevation was preceded by a transient HR drop. During 22-kHz USV presentation, a decrease in HR in both groups was observed, although it was more clear in paired-housed rats. CONCLUSIONS: Social context may have an impact on HR levels and USV emissions in response to ultrasounds presentations. However, it does not seem to influence the distinction between artificial and natural USV. FINANCIAL SUPPORT: The work was supported by National Science Centre, Poland, grant no. 2015/19/B/ NZ4/03393.
4

Irregular nuclear shapes of neural stem cells in adult murine dentate gyrus

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Grymanowska A. W., Martin A., Olszynski K. H., Wilczynski G. M., Magalska A., Filipkowski R. K.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: In adult hippocampal neurogenesis, stem cells and their derivative progenitors are generated. They differentiate into neurons as they migrate from the subgranular zone to the granule cell layer of the dentate gyrus, maintaining homeostatic tissue regeneration and supporting brain plasticity. Depending on the stage of the neurogenesis, different subpopulations of cells of the neurogenic lineage can be distinguished, i.e. neural stem cells (NSC, type 1), intermediate progenitor cells (type 2a and type 2b), neuroblasts (type 3) and granule neurons (type 4). Little is known about the architecture of nuclei in these cells, while the cell nucleus is known to be highly organized with numerous functional and structural domains as well as dynamic organization of chromatin governed by epigenetic mechanisms which were shown to respond to external signals. AIM(S): We aimed to distinguish type 1 through 4 cells and investigate their nuclear shape. METHOD(S): Transgenic Nestin-GFP mice were used. Cell types were identified with immunohistochemistry and morphological features: type 1 (GFP+, one long neural process), type 2a (GFP+), type 2b (GFP+/DCX+), type 3 (DCX+), type 4 (NeuN+). Confocal microscopy was used to collect z‑stack files of the nuclei of different cell types. RESULTS: We observed irregularity in shape of the nuclei in type 1 cells (NSC) with the presence of nuclear envelope invaginations. When selected layers were analyzed, NSC nuclei turned out to have reduced circularity, roundness and solidity when compared with other cell types. CONCLUSIONS: The irregularity observed and nuclear envelope invaginations seem to be characteristics of the “stemness” as the shape of the nucleus becomes more regular with successive stages of neurogenesis. The biological significance of the observed phenomenon is not yet clear and further studies are necessary to better understand the process of adult neurogenesis at the nuclear level. FINANCIAL SUPPORT: The work was supported by National Science Centre, Poland, grant no. 2014/14/M/NZ4/00561.
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Arginine and tetrahydrobiopterin supplementation in rats with salt-induced blood pressure increase: minor hypotensive effect but improvement of renal haemodynamics

61%
Kuczeriszka M., Walkowska A., Olszynski K. H., Rafalowska J., Sadowski J., Kompanowska-Jezierska E.
Journal of Physiology and Pharmacology
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2019
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tom 70
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nr 2
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