The differentiation of cerebellar interneurons is independent of their mitotic history.

A narrow time window centered around the terminal mitosis of their precursors has been recognized to be critical for the determination and/or realization of the developmental fate of a variety of neuronal phenotypes. In contrast, individual cell lineages in the cerebellum get separated early during embryonic development, and at least precursors for granule neurons have been found to be specified while still proliferating. We utilized primary dissociated cultures to address the issue of whether the faithful development of cerebellar granule cells and basket/stellate cells is dependent on their mitotic history and on the completion of a fixed number of cell cycles. Neuroblasts derived from embryonic cerebellar anlagen and transferred into primary dissociated cultures stopped proliferating as assessed by a loss of expression of the cell proliferation marker, Ki-67, and a failure to incorporate 5-bromo-2'-deoxyuridine. Although these cells had been forced to leave the proliferating cell pool prematurely, they developed into granule neurons or basket/stellate cells as judged by their distinct pattern of expression of specific molecular markers and the acquisition of a typical morphology. This included the cell intrinsic capacity of granule neurons to position their afferent synapses specifically to their dendrites. Thus, the competence of cerebellar interneurons to differentiate appropriately is independent of the precise timing of their final mitosis; however, their sensitivity towards extrinsic developmental signals appears to vary in a cell cycle-dependent manner, as suggested by the failure to survive of those cells that were in S-phase at the time of cultivation.