MITOCHONDRIAL RESPIRATION OF A PRIMARY MIXED CULTURE OF NEURONS FROM HIPPOCAMPUS AT VARIOUS STAGES OF DIFFERENTIATION

Abstract

BACKGROUND: Primary mixed culture of neurons is often used to evaluate the underlying molecular mechanisms of neurodegenerative disorders. However, the isolation of cells from the brain is accompanied by profound changes in cell morphology, behavior, metabolic rate due to enzymatic disintegration and changes in the microenvironment for cells. In this regard, it is relevant to study the basal respiration of mitochondria of neurons, as an indicator of the formation of the functional activity of the cell.AIM: to evaluate the mitochondrial respiration of a primary mixed culture of hippocampal neurons from embryos on the 18th day of gestation (E18) and 2-day-old pups at various stages of differentiation. METHODS: Primary embryonic (on the 18th day of gestation) and postnatal (on the 2nd day after birth) neuroglial culture of the CD1 mouse hippocampus was used in the work. The functional parameters of cell metabolism were measured by the Agilent analyzer (USA). The rate of the oxygen consumption was calculated based on the results of which of the profile of mitochondrial respiration of the primary mixed culture was built during its differentiation. RESULTS: Mitochondrial respiration in the postnatal hippocampus fully corresponds to the embryonic hippocampus. The oxidation rate increased almost 2-fold in the embryonic culture of hippocampal neurons, and an increase in metabolic potential was observed as cells differentiated in culture from 2 to 11 days. Neurons of the postnatal culture during differentiation showed a significant decrease in the rate of acidification of the medium by ~4.5 times against the background of a practically unchanged rate of oxidation, which indicates a decrease in the metabolic potential of the culture in the process of maturation. CONCLUSION: The results obtained prove that hippocampus can be used to study the role of mitochondria in neurogenesis.