Abstract: In order for the brain to develop with proper size and structure, neural stem cells (NSCs) at early
ages must make proliferative divisions and maintain their stemness to expand the stem cell pool, but then
switch to undergo neurogenic divisions at the correct time to create neurons. However, the mechanism of
this fate choice from remaining an NSC early on, to later choosing to exit the cell cycle and become a
neuron, is still poorly understood. The last step of cell division is abscission, which severs the daughter from
the mother cell. Abscission occurs during the time when the fate decision is made, and at the apical
membrane, where many fate signals are located. We developed methods and tools to quantitatively analyze
abscission in cortical NSCs, in vivo and in vitro. We found that abscission is not simply necessary to cut cells
apart and keep them alive. Rather, we made the surprising discovery that both abscission duration and
remnants of abscission (midbody remnants) are developmentally regulated, changing as development
proceeds. Furthermore, we found that a small-brained mouse mutant with altered abscission duration has a
reduced proportion of proliferative NSC divisions. These data led to our central hypothesis that changes in
abscission duration and midbody remnant persistence can shift NSC daughter cells fate choices as
development proceeds. We will test this hypothesis through the use of innovative genetic and cell biological
approaches, on single NSC divisions and whole tissue analyses. We will utilize two mouse mutants that
perturb abscission specifically, affecting duration and midbody remnants differentially. We will carry out three
Specific Aims: 1) test whether abscission duration is correlated with daughter cell fate outcomes in vitro, 2)
dissect the primary and secondary effects of dysregulated abscission on cortical NSC daughter cell fates,
morphologies and lineage progression in vivo, and 3) investigate a candidate signaling mechanism at the
apical membrane that could link abscission regulation to stem cell maintenance. The contributions of the
proposed research will be to increase understanding of the fundamental question of how stem cells in
developing tissues maintain high proliferative capacity early and then reduce it later in favor of differentiated
daughter cell types. It will also elucidate how regulation of NSC divisions affect daughter cell fates,
structures, and subsequent divisions. These contributions will be significant because they will reveal novel
mechanisms and gene pathways that regulate how brain size and structure are controlled, and will elucidate
how specific alterations in NSC division mechanisms during development can lead to brain malformations, or
other neurodevelopment phenotypes.