Cell Cycle-dependent Cues Regulate Temporal Patterning of the Central Brain Neural Stem Cells

During nervous system development, diverse types of neurons and glia are sequentially generated by self-renewing neural stem cells (NSCs). Temporal changes in gene expression within NSCs are thought to regulate neural diversity; however, the mechanisms regulating the timing of these temporal gene transitions remain poorly understood. type II NSCs, like human outer radial glia, divide to self-renew and generate intermediate neural progenitors, amplifying and diversifying the population of neurons innervating the central complex, a brain region required for sensorimotor coordination. Type II NSCs express over a dozen genes temporally, broadly classified as early and late-expressed genes. A conserved gene, Seven-up mediates early to late gene expression by activating ecdysone receptor (EcR) expression. However, what determines the timing of EcR expression and, hence, early to late gene transition is unknown. This study investigates whether intrinsic mechanisms of cell cycle progression and cytokinesis are required to induce the NSC early-late transition. By generating mutant clones that arrest the NSC cell cycle or block cytokinesis, we show that both processes are necessary for the early-to-late transitions. When NSCs are cell cycle or cytokinesis arrested, the early gene Imp failed to be down-regulated and persisted into the old NSCs, while the late factors EcR and Syncrip failed to be expressed. Furthermore, we show that the early factor Seven-up is insufficient to drive the transition despite its normal expression in the cell cycle- or cytokinesis-inhibited NSCs. These results suggest that both intrinsic (cell cycle/cytokinesis) and extrinsic (hormone) cues are required for the early-late NSC gene expression transition.