Adult neurogenesis is the lifelong generation of new neurons that occurs into restricted regions of the adult mammalian brain, namely the dentate gyrus of the hippocampus and the olfactory bulb. In this chapter, we describe the procedures aimed to investigate adult neurogenesis in the murine brain. Specifically, we explain how to inject into animals exogenous markers of proliferation (i.e., BrdU) and prepare brain tissues to perform immunofluorescence reactions for neuronal markers in combination with BrdU staining. As BrdU is incorporated in the DNA during the S-phase of the cell cycle of proliferating cells and is then inherited by daughter cells, by coupling BrdU-immunoreactivity together with the immunolabeling for neuronal markers, we provide the general procedures that can be applied to identify adult-born neurons and to characterize their specific phenotypes in different brain regions, under physiological condition or in pathological states.
Keywords: Adult neurogenesis; BrdU labeling; Multiple immunolabeling; Neuron birth-dating; Neuronal differentiation.