Single and Dual Birthdating Procedures for Assessing the Response of Adult Neural Stem Cells to the Infusion of a Soluble Factor Using Halogenated Thymidine Analogs

The factors that regulate the switch from adult neural stem cell (aNSC) quiescence to active proliferation are poorly understood. Here we describe a method to study the in vivo effect of a soluble factor on cell cycle entry and proliferation of aNSCs located in the brain neurogenic niches. First, we provide information for implanting osmotic minipumps that will deliver the compound of interest directly into the mouse brain. When combined with the administration of the thymidine analog bromodeoxyuridine (BrdU), this technique is the most basic procedure to study the effects of a soluble factor on aNSC proliferation. We also describe a dual replication labeling protocol using two different halogenated thymidine analogs, chloro- and iododeoxyuridine (CldU and IdU), that allows tracking of proliferating cells and assessing cell cycle re-entry of aNSCs at different time points. Curr. Protoc. Stem Cell Biol. 21:2D.10.1-2D.10.20. © 2012 by John Wiley & Sons, Inc. Keywords: neural stem cell; label-retaining cell; IdU; CldU; BrdU; intracerebroventricular infusion; osmotic minipump