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1

Selected behavioral and motor parameters in Bcl-2 knockout mice

100%
Slugocka A., Barski J. J.
Acta Neurobiologiae Experimentalis
|
2013
|
tom 73
|
nr Suppl.1
Bcl-2 knockout mice doesn’t produce Bcl-2 protein – one of the major protein factors regulating apoptotic processes involved in development of the nervous system. Bcl-2 deficiency result in increased cell death of motoneurons, sensory and sympathetic neurons (Michaelidis et al. 1996). Only very sparse data on impact of this mutation on motor activity and behavior are available. In the present study we examined the exploratory activity of knockout mice in the open field task, and checked motor related behaviors by means of RotaRod, elevated runway test, strength grip test, and balance rod test. Preliminary data suggest lack of prominent alteration of analyzed parameters, but more experiments are planned to look for possible more subtle changes.
2

Ultrasonic vocalization (USV) in calbindin D-28k mutant mice

81%
Grabowska M., Slugocka A., Barski J. J.
Acta Neurobiologiae Experimentalis
|
2013
|
tom 73
|
nr Suppl.1
Mice similarly to some other rodent species communicate with specialized sounds in the ultrasonic range called ultrasonic vocalizations (USV). Evaluation of this behavioral activity enables estimation of the social interactions in animal models of diseases involving psychiatric manifestations related to the social environment like autistic spectrum disorders (ASD). Because of the growing evidence for involvement of cerebellum in ethiology of ASD, we decided to change physiological properties of Purkinje cells and look for signs of alterations in USV activity of newborns. In our experiments we switched off expression of the major protein calcium buffer (calbindin D-28k) in these neurons by means of the Cre/loxP technology. It is known from previous reports (1), that the lack of calbindin D-28k in cerebellar cortex results in motor coordination deficits due to disturbed processing of calcium signaling in Purkinje cells. Because coordination of the vocal apparatus depends on the cerebellar input we were curious if altered Purkinje cells function results in altered USV calling.
3

Impact of ketogenic diet on gait in Purkinje cell specific transgenic mouse model of tuberous sclerosis complex

81%
Slugocka A., Grabowska M., Barski J. J.
Acta Neurobiologiae Experimentalis
|
2019
|
tom 79
|
nr Suppl.1
INTRODUCTION: The ketogenic diet is a high fat low carbohydrate diet, wherein the majority of caloric needs is covered by fats with very low carbohydrate intake. The diet is widely used not only by athletes and patients suffering from obesity or diabetes, but also by patients with intractable epilepsy. The high fat, low carbohydrate diet is extensively studied within the fields of numerous diseases including cancer and neurological disorders. METHOD(S): In present study, we used Purkinje cell (PCs) specific knockout mice lacking hamartin (tsc1), a key protein involved in mTORC1 pathway. Deletion of Tsc1 gene in PCs results in a loss of these cells and gait impairments. We implemented a ketogenic rodent chow to reveal its potential influence on prevention of loss of PCs in the cerebellum. We assessed the effects of treatment with the ketogenic diet on the quality of mice gait. The gait was analysed in the CatWalk system from Noldus. Obtained data were compared among groups: control animals, with tsc1 in PCs, fed with standard rodent diet, animals with tsc1 but fed with ketogenic chow and knockout mice, fed respectively with two types of abovementioned diets. RESULTS: Our results revealed that, as expected, animals without hamartin in PCs present severe gait disturbances. Supplementation of the ketogenic diet has no effect on gait disturbances caused by deletion of tsc1 in PCs. CONCLUSIONS: Additionally, statistical analysis of data obtained from animals without gene deletion didn’t bring any proof of differences in gait parameters between groups fed with two different chows.
4

Ultrasonic vocalization (USV) patterns in Eker rat newborns

81%
Slugocka A., Chojnacka G., Grabowska M., Barski J. J.
Acta Neurobiologiae Experimentalis
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2014
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tom 74
|
nr 3
Tuberous sclerosis complex (TSC) is one of the most frequent neurocutaneous disorders mostly present in pediatric patients. In TSC development we can observe neuropsychiatric signs with symptoms of epilepsy, mental retardation, and autism. Molecular mechanism of this disease involves two genes TSC1 and TSC2. Expression of both of them results in two proteins tuberin and hamartin negatively regulating mTOR signaling pathway. Mutations in TSC1 or TSC2 disturb their inhibitory function, what impairs the proliferation machinery downstream of mTOR. It results in growth of tumors (tubers) in many organs including the brain structure. Eker rat is a strain, that carries a spontaneously occurring germline mutation in the TSC2 gene. By the age of 1 year all TSC2Ek/- animals develop renal epithelial tumors. Tumors also appear in the brain tissue and make the strain a suitable model of autism spectrum disorders (ASD). One of the symptoms specific for autistic-like behaviors is the impaired communication between mother and child. Many rodent species including mouse and rat emit at the ultrasonic range (USV) sounds – a vocal communication between mother and offspring. This phenomenon is a good indicator of social interactions and alterations of the USV can be analyzed by means of specialized recording systems. In our study we attempted to check if there are any differences between TSC2Ek/- genotype newborns and wild type ones in patterns of USV using the standard isolation test. We analyzed sonographic structure of calls, number of calls per minute, duration of single call, etc.
5

Changes in the repetitive stereotyped characteristic of calcium binding proteins (CaBPs) knockout mice in the autistic-like behavior

81%
Grabowska M., Slugocka A., Barski J. J.
Acta Neurobiologiae Experimentalis
|
2014
|
tom 74
|
nr 3
Autism is a multifactorial disorder that involves impairments in social interactions and communication, as well as restricted and repetitive behaviors. Recent whole-genome exon sequencing studies of ASD (autism spectrum disorders) samples estimate that as many as 100 to 1 000 genes may be involved. So different candidate genes have been tested in mouse models by knocking them out. Calbindin D-28k (CB) and parvalbumin (PV) are cytosolic calcium-binding proteins expressed in many neurons without general preference for functionally and morphologically defined subpopulations. Deletion of CB and/or PV alters intracellular calcium signaling, and physiological properties of affected neurons. General knockouts for both proteins display a distinct and permanent motor impairment which is revealed only when adaptation of movement to novel environmental conditions is required. In order to determine whether the absence of CB and PV influences locomotor properties and behavior we compared mouse lacking CB, PV, or both with wild type controls. The mice were compared in the open field task and in the light-dark compartment tests to measure activity, exploratory behavior, and restricted and repetitive behaviors (TruScan, Coulbourn, USA). Relative to wild-type mice, the transgenic mice exhibited much more stereotypic movements in the open field test. Multiple mouse behavioral effects suggest that the CB and PV genes may play a role in modulating behaviors relevant to psychiatric disorders.
6

Ketogenic diet and sexual motivation in male rats

61%
Bogacki-Rychlik W., Bialy M., Grajoszek A., Przybyla M., Slugocka A., Barski J. J.
Acta Neurobiologiae Experimentalis
|
2019
|
tom 79
|
nr Suppl.1
INTRODUCTION: The ketogenic diet (KD) is a very‑low‑carbohydrate, high‑fat and adequate protein nutritional approach that induces metabolic shift to the use of ketone bodies instead of glucose as a main energy source. For decades, the KD has been employed to manage drug-resistant epilepsy, but recently it is increasingly considered as an alternative or add-on therapy in many other disorders. The positive effect of the KD on social behavior has been recently reported in rodent models of autism spectrum disorder (ASD) and wild type early adult male rats. AIM(S): Given, however, the influence of ketone bodies on many humoral parameters including testosterone, we decided to look closer at the sexual interactions of male rats subjected to the KD. METHOD(S): In our study, we examined behavior of males in response to female presence with concomitant analysis of ultrasonic vocalizations (USV) during sexual interactions. RESULTS: Percent of males starting to copulation significantly decreased during the second sexual session, which suggests lower sexual motivation in male rats on the KD. FINANCIAL SUPPORT: Funding: Ministry of Science and Higher Education, grant number KNW‑1‑053/N/8/I.
7

Ketogenic diets based on fat of either animal or plant origin have different effects on the abundance of autophagic vesicles in mouse brain

51%
Liskiewicz D., Liskiewicz A., Nowacka-Chmielewska M., Jablonska K., Grabowski M., Student S., Slugocka A., Przybyla M., Barski J. J., Malecki A.
Acta Neurobiologiae Experimentalis
|
2019
|
tom 79
|
nr Suppl.1
INTRODUCTION: Autophagy is a cellular recycling mechanism essential for maintenance of cell homeostasis and viability, especially during stress conditions; hence, autophagy is involved in a number of physiological and pathological processes. Autophagy is thought to be involved in anti‑aging and neuroprotective effects of caloric restriction, Sirtuin 1 activation, inhibition of insulin/ insulin-like growth factor signaling, and administration of rapamycin, resveratrol, and metformin. The ketogenic diet mimics the biochemical actions of fasting and exerts many physiological and cellular responses similar to those evoked by intermittent energy restriction. Despite this, the relationship between nutritional ketosis and autophagy has been a largely unexplored field. AIM(S): The aim of this study was to verify the hypothesis that ketogenic diets affect the process of autophagosome formation in the hippocampus and/or cerebral cortex. METHOD(S): 9-week-old male mice were fed with one of two differently composed ketogenic chows – based on the fat of either animal or plant origin (KA, KP respectively) or with standard rodent chow (SD) – for 6 subsequent weeks. Western blotting, (LC3, p62), QRT‑PCR (LC3A, LC3B, p62), and confocal microscopy (LC3 puncta) were employed to monitor autophagy in hippocampal and cerebrocortical samples. RESULTS: Western blot results revealed increased levels of LC3 II protein – a marker of autophagosomes – in the hippocampus and frontal cortex of mice treated with the ketogenic diet. This observation was confirmed by the evaluation of a number of LC3 puncta with immunofluorescence microscopy. The size of this effect was dependent on the composition of the diet. CONCLUSIONS: This study reports, for the first time, an upregulation of autophagosome synthesis in the brain of animals fed with the ketogenic diet. Our results make a significant contribution to the understanding of the mechanisms of ketogenic diet action. FINANCIAL SUPPORT: This research is supported by the National Science Center grant no. 2017/01/X/ NZ3/00984.
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