Extracellular nucleotides can control proliferation of neural stem cells (NSCs). However, their effects on selection of neuronal subtypes have not been elucidated. Glutamatergic neurons are the most abundant subtype in the mammalian brain. Production of this neuronal subtype can be observed not only in the development of the forebrain, but also through life in the hippocampus. In the development, glutamatergic neurons are produced from neuroepithelial cells at the dorsal side of the anterior neural tube. In the adult hippocampus, stem cells located at the sub-granular zone can produce glutamatergic granule neurons through life. We found that the expression of the nucleotide receptor P2Y4 was transiently augmented in the course of neuronal differentiation of mouse ES cells. Interestingly, a subpopulation of type 2 NSCs of the adult mouse hippocampus also expressed P2Y4. The activation of P2Y4 in those cells increased proportion of glutamatergic subtype in their descendant neurons. Our results provide evidence that differentiating NSCs pass through a stage in which nucleotides can affect subtype marker expression of their descendant neurons.