Wyniki wyszukiwania - Biblioteka Nauki

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Badanie mechanizmów zachowania zwierząt - testy behawioralne w naukach biomedycznych

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Boguszewski P. M.

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2011

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tom 48

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Social interaction in rodents - contemporary approach to animal behavior

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Boguszewski P. M.

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nr S

EN

Social behavior is a form of simple communication between members of the same species - both in humans and in animals. It seems to be a quite simple phenomenon, but due to its complexity and multidimensionality is difficult to quantify and analyze. In rodents and other animals social behavior constitutes a good model of the human interpersonal functioning. Various paradigms of social interaction in rats have been successfully used to study neuronal mechanisms of anxiety, aggression, domination, social defeat, stress, autism, individual differences in emotional reactivity as well as the effects of anxiogenic and anxiolytic drugs. A key advantage of this approach is a use of natural stimulus - another conspecific animal - instead of artificial objects or elaborated tasks. In addition animal reactions, that are observed as dependent variables, belongs to their natural repertoire. However, studies with multiple animals present the researcher with special challenges, both in experiment design, measurement techniques and in the analysis of data. Modern computer technology gives assistance to traditional, human observer based behavior coding as well as it does allow development of fully automatic behavior recognition systems. In this talk contemporary approach to social interaction in rodents will be presented along with major challenges and perspectives.

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Sexual behavior of male rats with moderate depression-a preliminary study

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Bialy M. , Nikolaev E. , Boguszewski P. M.

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nr 3

EN

Sexual behavior of male rats is well described type of appetitive behavior. This behavior is described by at least five independent factors: anticipatory, initiation, copulatory rate, hit rate and intromission count. Each factor has specific neuronal networks. The aim of our study was to analyze which factor is the most informative to describe sexual deficit in rats with depressive like syndrome. We investigate 6–8 month old WAG/Rij rats with absent epilepsy commodity with moderate depression and compare sexual behavior with control Crl:Han Wistar and Sprague-Dawley males. Sexually naïve WAG/Rij rats started to copulation less frequently (only 6 from 20 display mounting) compare to Crl:Han Wistar (6 from 6) and Sprague-Dawley (4 from 6). Sexually experienced WAG/Rij rats displayed longer mount latency (initiation factor) compare to both control strains. Surprisingly, sexually experienced WAG/Rij rats vocalized very intensively at 50-kHz band during five minutes before introduction of female. This anticipatory precontact vocalizations did not correlate with initiation of copulation. Latency to initiation of copulation (initiation factor) seems to be the most sensitive parameter describing depressive like syndrome during male rat sexual behavior and seems to be useful to investigation depressive disorders. It correspond to lower libido in men depressive patients.

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New and “cool” methods in behavioral neuroscience

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Boguszewski P. M. , Puscian A. , Knapska E.

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nr 3

EN

Behavioral tests in laboratory rodents play an essential role in basic and applied biomedical research. Development of new animal models for neurological and psychiatric disorders, as well as preclinical phase of drug research require a „proof-of-concept” testing on a system level. Since most of the behavioral procedures are not rigorously standardized, it is difficult to obtain replicable results between laboratories. One of the approaches to solve this problem is designing more ethologically-relevant behavioral tasks, in which behavioral expression is more voluntary and manifold behavioral measures are collected over long periods. Collecting large amounts of data requires automatic control of all stages of a study – experimental cage/system manipulation, data gathering and analysis. Such automated systems offer important advantages. They increase level of standardization that results in more coherent data, save time and manpower, as well as reduce animal numbers required. Automated monitoring, although new and often sophisticated, could be cheap as well. The inspiration comes from growing popularity of amateur robotics, accessibility of 3D printing and progress in electronics. The aim of the lecture is to present new tests and solutions we are developing in Nencki Institute, including RFID tags for social experiments, automatic vocalization classification, video image animals detection and recognition software and even Mindstorm Lego robots.

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Investigation of Ca2plus homeostasis and behavioral changes in transgenic mice overexpressing key store-operated calcium entry proteins

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Maciag F. , Majewski L. , Boguszewski P. M. , Kuznicki J.

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nr Suppl.1

EN

INTRODUCTION: Store-operated calcium entry (SOCE) is the major Ca2+ influx pathway in non-excitable cells. However, recent studies suggest its important roles in neurons. In SOCE, the depletion of Ca2+ from the endoplasmic reticulum (ER) causes an influx of Ca2+ from the extracellular space to refill the intracellular Ca2+ stores. STIMs are Ca2+ ER sensors that mediate SOCE by interacting with the ion channels in the cell membrane – ORAIs. Using transgenic mice with neuronal overexpression of STIMs and/or ORAIs, we investigate their role in neural function. Recently, we showed electrophysiological changes in hippocampi from female mice overexpressing ORAI1. Earlier studies from our group revealed increased cytoplasmic Ca2+ levels in cultured neurons overexpressing both ORAI1 and STIM2. Currently, we are extending these studies with the use of double transgenic STIM2/ORAI1 mice. AIM(S): To investigate the role of SOCE proteins in neurons, and the effect of STIM2 and ORAI1 overexpression on Ca2+ homeostasis, synaptic functions, and behavior. METHOD(S): We use transgenic mice that overexpress STIM and/or ORAI proteins in brain neurons. For studying Ca2+ homeostasis, we stain hippocampal slices with Fura‑2 AM probe. To assess locomotor functions and cognitive abilities of these mice, behavioral tests are utilized. Synaptic transmission and plasticity phenomena are investigated by electrophysiological recordings from hippocampal slices. RESULTS: We have recently observed spontaneous seizure-like events in aged female mice overexpressing ORAI1. These observations correlated with changes in the response of hippocampal slices to pro-epileptic drugs. Currently, we are focusing our analyses on the double transgenic STIM2/ORAI1 mouse line. CONCLUSIONS: Our previous data support the view that SOCE proteins play an important role in neurons. Currently, we aim to elucidate the involvement of STIM2 and ORAI1 proteins in neural function. FINANCIAL SUPPORT: Maestro to JK from NCN (2011/02/A/NZ3/00144).

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Neuroprotective effect of fingolimod and pramipexole in Parkinson's disease animal model

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Motyl J. , Przykaza L. , Kosson P. , Boguszewski P. M. , Strosznajder J. B.

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tom 75

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nr Supl.

EN

BACKGROUND AND AIMS: Recently sphingolipids alterations have been shown to play an important role in pathomechanism of neurodegenerative diseases. Our last study indicated suppression of gene expression and activity of sphingosine kinases (Sphk1/2)/ sphingosine-1-phosphate (S1P) synthesis in cellular model of Parkinson’s disease (PD). Moreover, the cytoprotective effect of S1P and its analog Fingolimod (P-FTY720) in this PD model was observed. The fundamental goal of current research was to determine the impact of FTY720 and dopamine D2/D3 receptors agonist – pramipexole (PPX) on death signalling and motor activity in mice PD model. METHODS: Neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 40 mg/kg) was administrated ip to adult C57BL/6 mice. FTY720 (1 mg/kg) or PPX (1 mg/kg) was injected ip during 10 days. Then behavioral tests (open field, rota-rod, and pole test) were performed. Midbrain and striatum were used for further studies. The immunochemical, spectrofluorometrical, and QPCR methods were applied. RESULTS: Our results indicated significant reduction in the number of dopaminergic cells in the midbrain of MPTP treated animals. Moreover, in this PD model alterations of Sphk1/2 and Akt kinase mediated signalling were found. It was also detected that gene expression of pro-apoptotic proteins in midbrain cells was activated. FTY720 and PPX protected dopaminergic cells against death as a result of Sphks up-regulation and apoptotic signalling suppression. In behavioural examination, MPTP mice exhibited impaired motor coordination in rota-rod test. Total time spent on the accelerating rota-rod was increased two-fold after FTY720 and PPX administration. We have also observed total distance elongation in animals treated with both above mentioned compounds during open-field test. CONCLUSIONS: In conclusion, this study indicated that FTY720 and PPX contribute to improvement of mice motor activity and they offer opportunities for PD therapy. Supported by NCN grant 2013/09/N/NZ4/02045.

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Maternal immune activation alters synaptic protein expression and causes social interaction impairment in rat offspring

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Cieslik M. , Gassowska-Dobrowolska M. , Jesko H. , Boguszewski P. M. , Filipkowski R. K. , Adamczyk A.

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nr Suppl.1

EN

INTRODUCTION: Emerging epidemiology data indicate that maternal immune activation (MIA) resulting from inflammatory stimuli such as bacterial infections during pregnancy may constitute a risk factor for multiple neurodevelopmental diseases including autism spectrum disorders (ASD). Genetic and environmental variation, inflammation during early development, and their interaction can influence synaptic dysfunction in ASD. However, the molecular links between infection-induced fetal development alterations and the risk of ASD are still unclear. AIM(S): The aim of this study was to investigate the effect of MIA on the expression and protein level of key synaptic proteins along with the autism-associated behavior in male rat offspring. METHOD(S): Pregnant Wistar rat dams were injected intraperitoneally (i.p.) at gestational day 9.5 with 0.1 mg/kg lipopolysaccharide (LPS), which induces immune response similar to that against gram-negative bacteria. RESULTS: Our data shown impaired social interaction, tested by the play behaviors (Tickling test on post-natal day PND 45–50). However, we did not observe any changes in ultrasonic vocalization (9–11 PND) and bedding preference (PND 15) in MIA offspring. Along with the social interaction changes, MIA has induced presynaptic protein alterations in adolescent rat offspring. These alterations included decreased level of synaptobrevin and syntaxin-1, the key components of SNARE complex, as well as higher level of synapsin. Together with changes in presynaptic proteins, MIA induced reduction in PSD-95 and down-regulation of SHANK family proteins. Moreover, alteration in the protein level of phospho-Akt, and 4E-BP1 was found in MIA subjects. CONCLUSIONS: It is possible that variations of Akt/ mTOR pathway are responsible for aberrant synthesis of key synaptic proteins. The altered synthesis of these proteins would generate changes in synaptic structure and function, contributing to ASD-like behaviors. FINANCIAL SUPPORT: Supported by the statutory theme 8.

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Sphingosine kinase-1, the new target in the neuroprotective effect of fingolimod and pramipexole in Parkinson’s disease animal model

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Motyl J. , Wencel P. L. , Przykaza L. , Kosson P. , Boguszewski P. M. , Strosznajder J. B.

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nr Suppl.1

EN

INTRODUCTION: Sphingosine kinase (Sphk1) synthetizing sphingosine-1-phoshate (S1P) is a key enzyme responsible for the regulation of cell fate. Sphk1/S1P could be the attractive target in Parkinson’s disease (PD) neuroprotective therapy. Our previous data showed inhibition of Sphk1 expression/activity in PD in vitro model and indicated neuroprotective effect of S1P analog phospho‑fingolimod (FTY720-P). AIM(S): The aim of current research was to investigate the effect of FTY720 and dopamine D2/D3 receptors agonist – pramipexole (PPX) on Sphk1 dependent molecular pathway(s) in selected parts of the brain and on locomotor activity in PD animal model. METHOD(S): Neurotoxin 1-methyl-4-phenyl-1,2,3,6- -tetrahydropyridine (MPTP, 40 mg/kg) was administrated i.p. to adult C57BL/6 mice. FTY720 (1 mg/kg) or PPX (1 mg/kg) were injected i.p. during 10 days. Behavioral tests (open field, rota-rod) were performed. Midbrain and striatum were separated. The immunochemical, spectrofluorometrical, and QPCR methods were applied. RESULTS: Our data indicated that PD mice exhibited significant loss of dopaminergic nerve terminals within striatum, evaluated by reduced tyrosine hydroxylase immunoreactivity level (TH-IR). Moreover we found the lower level of mRNA/ immunoreactivity and activity of Sphk1 in the midbrain of PD mice. Both FTY720 and PPX significantly increased TH-IR in MPTP mice striatum. FTY720 and PPX protected against MPTP-evoked Sphk1 alterations and significantly elevated pro-survival Akt kinase phosphorylation, which indicated its activation. Subsequently, FTY720 increased BAD protein phosphorylation in MPTP mice midbrain, which may protect cells against BAD-mediated death. Then it was observed that FTY-720 and PPX improved locomotor impairment in PD mice. CONCLUSIONS: Our data indicated the new neuroprotective mechanism of PPX and FTY720 action connected with sphingolipid signaling and demonstrated beneficial properties of these compounds on movement alterations in PD animal model. FINANCIAL SUPPORT: This abstract is financially supported by The National Science Centre grant 2013/09/N/ NZ4/02045.

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Antidepressant-like and anxiolytic-like effects of mild hypobaric hypoxia in mice: possible involvement of neuropeptide Y

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Duszczyk M. , Gamdzyk M. , Ziembowicz A. , Boguszewski P. M. , Lazarewicz J. W. , Salinska E.

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nr 4

EN

Previous studies have demonstrated that repeated submission of rats to mild hypobaric hypoxia reduces the persistent behavioral and hormonal depressive symptoms induced by exposure to footshock in the learned helplessness paradigm. The aim of this study was to determine whether hypoxic preconditioning of mice can also induce antidepressant- and anxiolyticlike effects that are detectable with the other commonly used behavioral tests, and to determine whether these effects are accompanied by an increase in neuropeptide Y (NPY) in the hippocampus, which may suggest the involvement of NPY in these mechanisms. The intermittent mild hypobaric hypoxia was generated by 2-h exposure of mice to 0.47 atm for 3 consecutive days. In the tail suspension test a significant decrease in the duration of immobility was observed 24 h, but not 48 h after the last hypobaric session. The elevated plus maze trials performed 48 h after preconditioning showed a significant increase in the frequency of open arm entries, a reduction in the duration of closed arm occupancy and substantially more time spent in the open arms in comparison to the control groups. The open field test demonstrated the absence of increases in general activity or unspecific exploratory behavior in hypoxia-preconditioned mice. The EIA test detected a statistically significant but relatively weak increase in the NPY content in the hippocampus 24 h after preconditioning. Together, our data demonstrate that preconditioning of mice with intermittent mild hypobaric hypoxia induces anxiolytic- and antidepressantlike effects. They are accompanied by up-regulation of NPY which may suggest its mechanistic role.

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Amotl1: a novel synaptic protein important for mice social behavior and brain organization

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Krzemien J. , Rojek K. , Winiarski M. , Boguszewski P. M. , Knapska E. , Proszynski T. J.

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nr Suppl.1

EN

INTRODUCTION: The angiomotin family comprises of three scaffold proteins – Amot, Amotl1, and Amotl2 – that have been implicated in the regulation of cell polarity, migration, and proliferation. Recent in vitro studies have reported that Amot localizes to the synapses in mature neurons and regulates dendritic spine maturation. AIM(S): We have found that Amot, together with Yap1, the Hippo pathway transcription co‑activator, are critical for proper dendritic arborization and mice locomotor coordination. However, to date the function of the two other Angiomotins, Amotl1 and Amotl2, in neurons has not been investigated. METHOD(S): To study Amotl1 function in the mouse brain, we generated systemic and neuron-specific knock‑out (KO) mice. To assess general locomotion, we performed an open field test. Amotl1 KO mice sociability was evaluated with the three-chamber task, automatic Eco‑Hab approach, and nesting test. To record the animal’s anxiety response, we used the marble burying test. RESULTS: In the present study, we show that Amotl1 localizes to the synaptic compartments in neurons. Deletion of Amotl1 leads to hyperlocomotion, decreased anxiety-like behavior, and alteration in mice sociability. Amotl1 ablation causes an increase in volume of lateral ventricles in the mouse brain by 50%. These features have been previously observed in animal models of various psychiatric disorders, such as schizophrenia or autism. Interestingly, mass spectrometry analysis of neuron‑specific interactors demonstrated that Amotl1 binds to FMR1 and FXR1, mutations of which cause Fragile X syndrome. CONCLUSIONS: We identified a novel synaptic protein, Amotl1, the deletion of which causes behavioral deficits and that it could be a potential molecular target for the development of new therapeutics for neurological disorders. FINANCIAL SUPPORT: This research was supported by National Science Center (NCN) grants: UMO- ‑2018/29/B/NZ3/02675, UMO-2018/29/N/NZ3/02682.