i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto 4200-135, Portugal.
IBMC-Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto 4200-135, Portugal.
Genetics. 2024 Mar 6;226(3). doi: 10.1093/genetics/iyae005.
The microtubule motor dynein is critical for the assembly and positioning of mitotic spindles. In Caenorhabditis elegans, these dynein functions have been extensively studied in the early embryo but remain poorly explored in other developmental contexts. Here, we use a hypomorphic dynein mutant to investigate the motor's contribution to asymmetric stem cell-like divisions in the larval epidermis. Live imaging of seam cell divisions that precede formation of the seam syncytium shows that mutant cells properly assemble but frequently misorient their spindle. Misoriented divisions misplace daughter cells from the seam cell row, generate anucleate compartments due to aberrant cytokinesis, and disrupt asymmetric cell fate inheritance. Consequently, the seam becomes disorganized and populated with extra cells that have lost seam identity, leading to fatal epidermal rupture. We show that dynein orients the spindle through the cortical GOA-1Gα-LIN-5NuMA pathway by directing the migration of prophase centrosomes along the anterior-posterior axis. Spindle misorientation in the dynein mutant can be partially rescued by elongating cells, implying that dynein-dependent force generation and cell shape jointly promote correct asymmetric division of epithelial stem cells.
微管马达动力蛋白对于有丝分裂纺锤体的组装和定位至关重要。在秀丽隐杆线虫中,这些动力蛋白的功能在早期胚胎中得到了广泛研究,但在其他发育环境中仍未得到充分探索。在这里,我们使用一个低功能的动力蛋白突变体来研究马达在幼虫表皮中不对称干细胞样分裂中的贡献。在 seam 细胞分裂之前的活细胞成像,这些 seam 细胞分裂先于 seam 合胞体的形成,表明突变细胞正确地组装,但经常错误地定位它们的纺锤体。错位分裂会导致子细胞从 seam 细胞列中错位,由于异常胞质分裂产生无核区,并破坏不对称细胞命运的遗传。结果,seam 变得混乱,并充满了失去 seam 身份的额外细胞,导致致命的表皮破裂。我们表明,动力蛋白通过沿前后轴引导前期中心体的迁移,通过 GOA-1Gα-LIN-5NuMA 皮层途径来定位纺锤体。动力蛋白突变体中的纺锤体错位可以通过延长细胞部分得到挽救,这意味着动力蛋白依赖性力的产生和细胞形状共同促进上皮干细胞的正确不对称分裂。