Neurogenesis – In Vivo Neurogenesis – Survival and Differentiation
Survival and differentiation of progenitor cells into neurons can be measured using a pulse-chase protocol in which animals receive concurrent drug treatment and 2H2O label and are sacrificed 4 weeks after the end of treatment. GC/MS analysis of anti-tetrodotoxin labeled neuronal cells or NeuN labeled nuclei can be used to determine changes in neurogenesis.
Mice were treated with fluoxetine and concurrently received 2H2O in the drinking water and were sacrificed 4 weeks after the end of treatment. Top left: Hippocampal neuronal cells were isolated by fractionation, labeled with tetanus toxin C fragment (TTX), and analyzed by flow cytometry. Top right: GC/MS analysis indicates that the fraction of label-retaining neuronal cells is significantly greater in fluoxetine-treated animals. Bottom left: Hippocampal nuclei were isolated, labeled with Neu-N and sorted by flow cytometry. Bottom right: GC/MS analysis indicates that the fraction of label-retaining neuronal nuclei is greater in fluoxetine-treated animals. ** indicates p < .01, *** indicates p<0.001, n = 8.
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Rats were treated with sodium valproate for 4 weeks and concurrently labeled with 2H2O for the last 3 weeks of drug treatment. Progenitor cells and neuroblasts were isolated from the subventricular zone (SVC) at the end of treatment. The olfactory bulb (OB) was isolated 2 weeks after treatment to assess neuronal differentiation. Sodium valproate treatment significantly increased neurogenesis by increasing both proliferation and differentiation. Bars indicates Mean + S.D., * p < .05
