Evaluation of Adult Mouse Brain Neurogenic Niche Behavior Culturing Adult Mice Brain Slice In Vitro.

Abstract

Adult brain neural precursors carry out their biological activity in specific areas in which they are able to self-renew and differentiate into neurons. This is due to a complex microenvironment of cellular interrelations in which soluble factors from the neighboring cells, vascular structures, and the content of the brain ventricle cavity (cerebrospinal fluid) play a key role. This cellular functional entity, known as the "neurogenic niche," is able to generate new mature neurons, which are functionally integrated into the neuronal circuits of the adult mammal brain. The complexity of neurogenic niche signaling, which include biologically active molecules such as growth factors and morphogens, requires an experimental approach in order to create specific modifications of the biological activity of some of these molecules by means of a model of the active neurogenic niche, allowing an evaluation of neural precursor behavior.Here we describe the adaptation of an in vitro culture technique of adult brain slices with selected coronal sections, involving the two main brain neurogenic niches, the sub-ventricular zone (SVZ), and the hippocampus dentate gyrus, together with their associated sub-ependymal zone (SEZ). We explain certain examples of the experimental approach to modify neurogenic niche soluble signaling, implanting latex microbeads as a carrier for soluble signals. Additionally, we introduce an immune-cytochemical approach involving bromodeoxyuridine detection as a neural precursor cellular lineage tracer in combination with different molecular expressions, as a means of testing progressive states of neural precursor differentiation and neuronal maturation.This system represents a suitable strategy for evaluating the biological role of soluble components of the adult brain neurogenic niche.