Eckerle Stephanie, Ringler Mario, Lecaudey Virginie, Nitschke Roland, Driever Wolfgang
Developmental Biology, Institute Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, Hauptstrasse 1, D-79104 Freiburg, Germany.
Developmental Biology, Institute Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, Hauptstrasse 1, D-79104 Freiburg, Germany; BIOSS - Centre for Biological Signalling Studies, Albertstrasse 19, 79104 Freiburg, Germany.
Dev Biol. 2018 Feb 15;434(2):249-266. doi: 10.1016/j.ydbio.2017.12.016. Epub 2017 Dec 26.
Control of microtubule dynamics is crucial for cell migration. We analyzed regulation of microtubule network dynamics in the zebrafish yolk cell during epiboly, the earliest coordinated gastrulation movement. We labeled microtubules with EMTB-3GFP and EB3-mCherry to visualize and measure microtubule dynamics by TIRF microscopy live imaging. Yolk cell microtubules dynamics is temporally modulated during epiboly progression. We used maternal zygotic Pou5f3 mutant (MZspg) embryos, which develop strong distortions of microtubule network organization and epiboly retardation, to investigate genetic control of microtubule dynamics. In MZspg embryos, microtubule plus-end growth tracks move slower and are less straight compared to wild-type. MZspg embryos have altered steroidogenic enzyme expression, resulting in increased pregnenolone and reduced progesterone levels. We show that progesterone positively affects microtubule plus-end growth and track straightness. Progesterone may thus act as a non-cell-autonomous regulator of microtubule dynamics across the large yolk cell, and may adjust differing demands on microtubule dynamics and stability during initiation and progression phases of epiboly.
微管动力学的控制对细胞迁移至关重要。我们分析了斑马鱼卵黄细胞在原肠胚形成(最早的协调原肠胚运动)过程中微管网络动力学的调控。我们用EMTB - 3GFP和EB3 - mCherry标记微管,通过全内反射荧光显微镜实时成像来可视化和测量微管动力学。在原肠胚形成过程中,卵黄细胞微管动力学在时间上受到调节。我们使用母源合子Pou5f3突变体(MZspg)胚胎,其微管网络组织出现严重扭曲且原肠胚形成延迟,以研究微管动力学的遗传控制。在MZspg胚胎中,与野生型相比,微管正端生长轨迹移动较慢且较不笔直。MZspg胚胎的类固醇生成酶表达发生改变,导致孕烯醇酮增加和孕酮水平降低。我们发现孕酮对微管正端生长和轨迹笔直度有正向影响。因此,孕酮可能作为一种非细胞自主的调节剂,作用于整个大的卵黄细胞中的微管动力学,并可能在原肠胚形成的起始和进展阶段调节对微管动力学和稳定性的不同需求。