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The capacity of neural stem cells (NSC) to generate different types of neurons and glia depends on the action of intrinsic determinants and extracellular signals. Here, we isolated adult olfactory bulb stem cells (aOBSC) that express nestin, RC2 and Sox2, and that have the capacity to generate neurons possessing mature features in culture and in vivo. The differentiation of aOBSC into neurons and glia, as well as their genetic profile, was compared to that of embryonic OBSC (eOBSC) and ganglionic eminence stem cells (GESC). While these eOBSC express neurogenin (Ngn) 1 and 2, two telencephalic dorsal markers, GESC only express Ngn2. Adult OBSC express either little or no detectable Ngn1 and 2, and they produced significantly fewer neurons in culture than eOBSC. By contrast, Dlx2 transcripts (a telencephalic ventral marker) were only clearly detected in GESC. When transplanted into the early postnatal P5-P7 OB, each of the three populations gave rise to cells with a distinct pattern of neuronal migration and/or dendritic arborization. Overall, these findings suggest that cultured NSC partially maintain their regional and temporal specification. Notably, significant neuronal migration and differentiation were only observed in vivo when the NSC were briefly exposed to fibroblast growth factor-2 (FGF-2) before grafting, a treatment that enhanced the neurogenin expression. Hence, the migration and maturation of neurons derived from transplanted NSC can be promoted by upregulating neurogenic gene expression with FGF-2.

Neural stem cells depend on insulin-like growth factor I (IGF-I) for differentiation. We analysed how activation and inhibition of the PI 3-kinase-Akt signalling affects the number and differentiation of mouse olfactory bulb stem cells (OBSCs). Stimulation of the pathway with insulin and/or IGF-I, led to an increase in Akt phosphorylated on residues Ser473 and Thr308 (P-Akt(Ser473) and P-Akt(Thr308), respectively) in proliferating OBSCs, and in differentiating cells. Conversely, P-Akt(Ser473) levels decreased by 50% in the OB of embryonic day 16.5-18.5 IGF-I knockout mouse embryos. Overexpression of PTEN, a negative regulator of the PI 3-kinase pathway, caused a reduction in the basal levels of P-Akt(Ser473) and P-Akt(Thr308) and a minor reduction in IGF-I-stimulated P-Akt(Ser473). Although PTEN overexpression decreased the proportion of neurons and astrocytes in the absence of insulin/IGF-I, it did not alter the proliferation or survival of OBSCs. Accordingly, overexpression of a catalytically inactive PTEN mutant promoted OBSCs differentiation. Inhibition of PI 3-kinase by LY294002 produced strong and moderate reductions in IGF-I-stimulated P-Akt(Ser473) and P-Akt(Thr308), respectively. Consequently, LY294002 reduced the proliferation of OBSCs and the number of neurons and astrocytes, and also augmented cell death. These findings indicate that OBSC differentiation is more sensitive to lower basal levels of P-Akt than proliferation or death. By regulating P-Akt levels in opposite ways, IGF-I and PTEN contribute to the fine control of neurogenesis in the olfactory bulb.