Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, VA 22908.
Neuroscience Graduate Program, University of Virginia School of Medicine, Charlottesville, VA 22908.
Proc Natl Acad Sci U S A. 2020 Apr 28;117(17):9584-9593. doi: 10.1073/pnas.1919658117. Epub 2020 Apr 9.
Building a brain of the proper size and structure requires neural stem cells (NSCs) to divide with tight temporal and spatial control to produce different daughter cell types in proper numbers and sequence. Mammalian NSCs in the embryonic cortex must maintain their polarized epithelial structure as they undergo both early proliferative divisions and later neurogenic divisions. To do this, they undergo a polarized form of cytokinesis at the apical membrane that is not well understood. Here, we investigate whether polarized furrowing and abscission in mouse NSCs are regulated differently at earlier and later stages and in a cytokinesis mutant, This mutant was previously shown to have microcephaly and elevated apoptosis of NSCs. We developed methods to live image furrow ingression and midbody abscission in NSCs within cortical explants. We find that polarized furrow ingression occurs at a steady rate and completes in ∼15 min at two different ages. However, ingression is slower in a subset of mutant NSCs. Abscission is usually observed on both sides of the midbody and takes 65 to 75 min to complete. Surprisingly, abscission is accelerated in the mutant NSCs. Postabscission midbody remnants are observed at the apical membranes of daughter cells and are much more abundant in early-stage cortices. After NSC divisions in vitro, midbody remnants are more often retained on the daughter cells of early proliferative divisions. Altogether, these results suggest that regulation of abscission timing and midbody remnants in embryonic NSCs may influence proper brain growth and structure.
构建大小和结构适当的大脑需要神经干细胞(NSCs)进行精确的时空控制分裂,以产生适当数量和顺序的不同子细胞类型。胚胎皮层中的哺乳动物 NSCs 在经历早期增殖分裂和后期神经发生分裂时,必须保持其极化的上皮结构。为了做到这一点,它们在顶端膜经历一种未被很好理解的极化胞质分裂形式。在这里,我们研究了在早期和晚期以及在胞质分裂突变体中,极化的沟和分裂是否在小鼠 NSCs 中受到不同的调节,这个突变体先前被证明具有小头畸形和 NSCs 的凋亡增加。我们开发了在皮质外植体中活细胞成像 NSCs 中沟的侵入和中体的分裂的方法。我们发现,极化的沟侵入以稳定的速度发生,在两个不同的年龄大约需要 15 分钟完成。然而,在一部分 突变体 NSCs 中,侵入速度较慢。分裂通常在中体的两侧观察到,需要 65 到 75 分钟才能完成。令人惊讶的是,在 突变体 NSCs 中,分裂被加速了。分裂后中体的残余物在子细胞的顶端膜上观察到,在早期皮质中更为丰富。在体外 NSC 分裂后,中体残余物更经常保留在早期增殖分裂的子细胞上。总之,这些结果表明,胚胎 NSCs 中分裂时间和中体残余物的调节可能影响大脑的正常生长和结构。