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. On the 14th day of the cultivation, the cells exposed to hypoxia. 1ng / ml BDNF was preemptively added in the examined cultures. To measure the functional activity of the hippocampal cultures RNA detection probe SmartFlare was used. To assess the changes in the functional activity of the 1st day after the simulation hypoxia detection of mRNA BDNF was carried out.For detection we used RNA probe SmartFlare, whose fluorescence was determined as helium-neon laser with λ=543. During examination of the percentage BDNF mRNA-positive cells in primary cultures of dissociated hippocampal cultures between 7 and 14 days of development (Fig. 1) we found a significant increase of BDNF mRNA-positive cell group which preventively got BDNF, relatively to the control group. There also a slight increase in mRNA BDNF-positive cells relatively to controls at 21 days of development, but no significant differences were found. During analyzing the changes in mRNA BDNF-positive cells in the temporal dynamics, we found that the proportion of BDNF mRNA was significantly increased at the 14th day of development in comparison with 7 days. Next 21 hours of significant drop of the mRNA BDNF. Statistical differences between 7 and 21 days was found. These data suggests that the preventive addition of BDNF percentage to dissociated primary hippocampal cultures on the 14th day of the development affects the synthesis of endogenous BDNF in the most positive way.