Brain cell composition analysis through the isotropic fractionator method in a murine experiment of maternal malnutrition

BARBEITO-ANDRES J; CASTRO E; BERNAL V.; HALLGRIMSSON B; LENT R; GONZALEZ PN

Buenos Aires

Congreso; Second FALAN congress; 2016

Nutrition is one of the environmental factors that mostly affects brain development. Different behavioral, cognitive, physiological and anatomical impairments are associated with nutritional restriction during early life. However, it is still largely unknown if changes in quantitative cellular composition (number, cell type proportions) in different brain regions occur as a result of maternal malnutrition. Using a mouse experimental model of maternal protein and caloric undernutrition, we assessed absolute cell number in six brain regions of the offspring by means of the isotropic fractionator method. This method consists on breaking the cell membranes and releasing the nuclei into a fluid of known volume. From this isotropic solution, aliquots are extracted and the density of cells is determined. All nuclei were stained and counted by means of DAPI while the proportion of neurons was determined using NeuN. Results indicated a significant reduction of the total number of cells in the cerebellum and the cortex under protein restriction. Changes in number of neurons varied across regions. In sum, this study enlarges our knowledge on brain susceptibility to disrupting factors during brain development.