Opera Medica et Physiologica

E.V. Mitroshina

Neuroprotective and Antihypoxic Effects of Glial Cell Line-Derived Neurotrophic Factor (Gdnf) in Hypoxia Modeling

INTRODUCTION

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Abstract

The aim of the investigation was to assess antihypoxic and neuroprotective properties of the glial cell line-derived neurotrophic factor (GDNF) in hypoxia models in vitro and in vivo. In vitro experiments were carried out on primary hippocampal cultures. Hypoxia modeling was performed on day 14 of culture development in vitro (DIV) by replacing the normoxic cultural medium with a medium containing low oxygen for 10 minutes. Registration of extracellular action potentials was conducted by MEA systems (Multichannel Systems, Germany) application. Study the effect of GDNF on synaptic plasticity was performed using SmartFlare RNA Detection Probes (Merck Millipore, France) and fluorescent microscopy. In vivo experiments were carried out on C57BL/6j male mice. For acute hypobaric hypoxia a vacuum flow-through chamber was used at the ambient temperature of 20–22°C. We have investigated the resistance of animals to hypoxia and their spatial memory retention in Morris water maze test 24 hours after hypoxia. In vitro and in vivo data demonstrated that GDNF has strong antihypoxic and neuroprotective properties. Preventive GDNF application before hypoxia contributed to the animal survival and spatial memory retention as well as the maintenance of cells viability in primary hippocampal cultures. 

The Role of Cannabinoid Receptors (Type 1 and Type 2) in Implementation of Neuroprotective and Antihypoxic Effects of N-Arachidonoyldopamine in Acute Hypoxia In Vitro

INTRODUCTION

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Abstract

The aim of the investigation was to study a role of cannabinoid receptors type 1 (СВ1) and type 2 (СВ2) in implementation of antihypoxic and neuroprotective effects of N-ADA in hypoxia model in vitro. The experiments were carried out on primary hippocampal cultures. N-ADA effect on the spontaneous bioelectrical and calcium network activity in dissociated hippocampal cultures in normal and hypoxic conditions as well as the role of CB1 and CB2 in the implementation of these effects were investigated. Registration of extracellular action potentials was conducted by MEA systems (Multichannel Systems, Germany) application. For the detection of patterns of spontaneous calcium oscillations we used fluorescent calcium dye Oregon Green 488 BAPTA-1 AM (Invitrogen) and a confocal laser scanning microscope (Zeiss LSM510, Germany). Study the expression of mRNA CB1 receptors was performed using SmartFlare RNA Detection Probes (Merck Millipore, France) and fluorescent microscopy. Our data demonstrated that N-ADA has strong antihypoxic and neuroprotective properties associated with activation of cannabinoid receptors type 1. 

The Influence Of Brain-Derived Neurotrophic Factor (Bdnf) On Functional Activity Of The Culture Hippocampus During Hypoxia (In Vitro Modelling)

Oxygen deficiency is the major cause of cell death at a large range of pathologies. The neurons are among body cells, which are the most sensitive to lack of oxygen, concerning the problem of brain hypoxia retains emergency medical and biological significance. The purpose of research is studying the impact of brain-derived neurotrophic factor (BDNF) on the functional activity of dissociated cultures of hippocampus in modeling normobaric hypoxia. In the in-vitro study we used dissociated hippocampal cell cultures derived from CBA mice 18 day embryos.

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