Restriction on self-renewing asymmetric division is coupled to terminal asymmetric division in the Drosophila CNS

Neuronal precursor cells undergo self-renewing and non-self-renewing asymmetric divisions to generate a large number of neurons of distinct identities. In Drosophila, primary precursor neuroblasts undergo a varying number of self-renewing asymmetric divisions, with one known exception, the MP2 lineage, which undergoes just one terminal asymmetric division similar to the secondary precursor cells. The mechanism and the genes that regulate the transition from self-renewing to non-self-renewing asymmetric division or the number of times a precursor divides is unknown. Here, we show that the T-box transcription factor, Midline (Mid), couples these events. We find that in mid loss of function mutants, MP2 undergoes additional self-renewing asymmetric divisions, the identity of progeny neurons generated dependent upon Numb localization in the parent MP2. MP2 expresses Mid transiently and an over-expression of mid in MP2 can block its division. The mechanism which directs the self-renewing asymmetric division of MP2 in mid involves an upregulation of Cyclin E. Our results indicate that Mid inhibits cyclin E gene expression by binding to a variant Mid-binding site in the cyclin E promoter and represses its expression without entirely abolishing it. Consistent with this, over-expression of cyclin E in MP2 causes its multiple self-renewing asymmetric division. These results reveal a Mid-regulated pathway that restricts the self-renewing asymmetric division potential of cells via inhibiting cyclin E and facilitating their exit from cell cycle.Author summary: Nerve cells in the brain, spinal cord, gut and so on in all organisms are generated from stem cells. These primary cells divide to self-renew and at the same time generate a secondary precursor cell that terminally divides to produce two cells that differentiate into neurons of different identities, or glial cells or a neuron and a glia. The secondary cells never self-renew, the reason for which is not known. We found that in embryos that lack the activity of a gene called midline, precursors such as MP2 that normally divides into two neurons, self-renews and generates a neuron at the same time. The identity of the differentiating progeny is tied to how the asymmetrically localized determinant Numb is distributed in the precursor cell. When this gene, midline, is over expressed, it blocks MP2 division. The way Midline protein works is that it represses the cyclin E gene via binding to sites in its promoter, preventing the over-expression of Cyclin E and thus blocking cells from entering the cell cycle. A deregulation of cyclin E as in loss of function midline mutants allows one of the daughter cells of MP2 to re-enter cell cycle as MP2, just as an over-expression of the cyclin E gene also does. These results show a mechanism by which restriction on self-renewing asymmetric division is coupled to terminal asymmetric division and works through Midline and Cyclin E. This work addresses one of the fundamental problems is biology.