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Volga Neuroscience School 2016 Astroglial control of rhythm genesis in the brain
The Effects of Caloric Restriction and Western Diet on Astrocytes
A. Lebedeva*, A. Plata, V. Tovpyga, P. Denisov, S. Makovkin, A. Semyanov
Institute of Neuroscience, Lobachevsky State University of Nizhny Novgorod, Nizhny Novgorod, Russia. * Presenting e-mail: lebedeva@neuro.nnov.ru
The study of the processes involved in aging has ample importance in neurobiology. Life expectancy has augmented in developed countries with the consequent increase in the incidence of neurodegenerative diseases. Preliminary studies have shown that caloric restriction diet can improve memory in the elderly. Furthermore, high-fatty (Western) diet can contribute to the development of neurodegenerative disorders such as Alzheimer's disease. Most of the recent works have involved different mechanisms of neural networks in aging and neuropathological processes. However, the role of neuroglia has been less frequently studied. In order to elucidate the role of astrocytes in aging, we studied how low caloric and high fatty regimens alter astroglia activity over time. We used male mice C57/BL/6 of different postnatal days (P), slices were taken at day 1-3, 5, 7, 14 and 30 after the beginning of a specific diet. Caloric restriction diet consists of lowering to 70% the standard ration of the animals. Western diet contained 50% more fat than in the standard menu. The control mice receive food ad libitum in compliance with the standards stipulated for laboratory animals. Currents from astrocytes of hippocampal slices were measured by patch clamp technique in physiological stimulation and subsequently under pharmacological blockade of the excitatory amino acid transporters (EAATs) with TBOA (50 |oM).
Acknowledgements
This work was supported by the Russian Science Foundation (project 16-14-00201).
GDNF Influence on the Morpho-Structural Integrity of Hippocampal Neural Network in Hypoxia Modeling In Vitro
T.V. Shishkina12 *, T.A. Mishchenko2,1, E.V. Mitroshina2,1, I.V. Mukhina2A, V.B. Kazantsev1, M.V. Vedunova12
1 The Institute of Biology and Biomedicine, Lobachevsky State University of Nizhny Novgorod, Nizhny Novgorod, Russia;
2 Molecular and cell technologies group, Nizhny Novgorod State Medical Academy, Nizhny Novgorod, Russia. * Presenting e-mail: schischkina.tatiana2012@yandex.ru
The preservation of neural networks' functional integrity under the influence of various stress factors is one of the topical issues in modern neurobiology. Neural networks are regarded as a minimal functional unit of the central nervous system, responsible for the transmission and information storage processes. Stressogenic factors, such as hypoxia, stimulate not only the death of the functionally important neurons, but reorganize the synaptic plasticity processes, which significantly affects the neural network functional structure. Nowadays, a substantial question, concerning the exploration of substances, able to maintain the viability and functional activity of brain cells in stressful conditions, remains open. Particular attention is given to investigation of neurotrophic factors, especially Glial cell line-derived neurotrophic factor (GDNF), which regulate various cellular processes during development and in mature brain. The one of the unique properties, that distinguishes this protein from other neurotrophins, is its ability to affect the synaptic plasticity by influencing on promoter activity of GluR2-subunit of AMPA-receptors.
In this regard, the aim of the investigation is to study the GDNF influence on viability and spontaneous bioelectrical activity of neural networks in primary hippocampal cultures during hypoxia modeling in vitro.
Materials and Methods
Dissociated hippocampal cells were taken from C57Bl6 mice embryos (E18) and cultured during 14 days in vitro according to the previously developed protocol on multielectrode arrays (Alpha Med Science, Japan) or coverslips. Hypoxia modeling was performed on DIV14 by replacing the normoxic cultural medium with a medium containing low oxygen for 10 min. The main parameters of spontaneous bioelectrical activity such as the number of bursts, the number of spikes in a burst, the burst duration were established. In addition, the method of correlation graphs was used. We also conducted the cell viability detection. Moreover, SmartFlareTM RNA Detection Probes (Merck Milli-pore, SFC-534, France) were applied for intravital detection of mRNA GluR2 in primary hippocampal cultures. The carried out experiments revealed an acute increase in the number of dead cells (4,5 times) in primary hippocam-
OM&P
112 Opera Med Physiol 2016 Vol. 2 (S1)
