Rose K L, Winfrey V P, Hoffman L H, Hall D H, Furuta T, Greenstein D
Department of Cell Biology, Vanderbilt University School of Medicine, 21st and Garland, Nashville, Tennessee 37232-2175, USA.
Dev Biol. 1997 Dec 1;192(1):59-77. doi: 10.1006/dbio.1997.8728.
In Caenorhabditis elegans, specialized contractile myoepithelial cells of the somatic gonad, the gonadal sheath cells, are closely apposed to oocytes and are required for normal meiotic maturation and ovulation. Previously we found that mutations in the ceh-18 gene, which encodes a POU-class homeoprotein expressed in sheath cells, result in oocyte defects. To determine the basis for these oocyte defects, we have used time-lapse video Nomarski microscopy to observe meiotic maturation, ovulation, and early embryogenesis in ceh-18 mutants. In ceh-18 mutants sheath cell contractions are weaker, less frequent, and uncoordinated throughout the sequence of ovulation events, and ovulation is defective. Defective ovulation can result in the formation of endomitotic oocytes in the gonad, the formation of haploid embryos, and reversals in embryonic polarity. ceh-18 mutant oocytes exhibit defects prior to nuclear envelope breakdown, suggesting that they are physiologically different from the wild type. We observed delays in meiotic maturation, as well as maturation out of the normal spatial and temporal sequence, suggesting that proximal sheath cells directly or indirectly promote and spatially restrict meiotic maturation. Analysis of sheath cell differentiation in ceh-18 mutants using antibodies to proteins of the contractile apparatus reveals that although contractile proteins are expressed, the sheath cells appear disorganized. Transmission electron microscopy reveals that ceh-18 mutant sheath cells are morphologically irregular and only loosely cover oocytes. Taken together, these observations indicate that ceh-18 is a crucial determinant of sheath cell differentiation, a function required for normal meiotic maturation and ovulation.
在秀丽隐杆线虫中,体细胞性腺的特化收缩性肌上皮细胞,即性腺鞘细胞,与卵母细胞紧密相邻,是正常减数分裂成熟和排卵所必需的。此前我们发现,编码在鞘细胞中表达的POU类同源蛋白的ceh-18基因突变会导致卵母细胞缺陷。为了确定这些卵母细胞缺陷的原因,我们使用延时视频Nomarski显微镜观察了ceh-18突变体中的减数分裂成熟、排卵和早期胚胎发育。在ceh-18突变体中,鞘细胞收缩较弱、频率较低,并且在整个排卵事件序列中不协调,排卵存在缺陷。有缺陷的排卵会导致性腺中形成核内有丝分裂卵母细胞、单倍体胚胎的形成以及胚胎极性的反转。ceh-18突变体卵母细胞在核膜破裂之前就表现出缺陷,这表明它们在生理上与野生型不同。我们观察到减数分裂成熟存在延迟,以及成熟偏离正常的时空序列,这表明近端鞘细胞直接或间接促进并在空间上限制减数分裂成熟。使用针对收缩装置蛋白的抗体对ceh-18突变体中的鞘细胞分化进行分析,结果显示尽管收缩蛋白表达,但鞘细胞看起来杂乱无章。透射电子显微镜显示,ceh-18突变体鞘细胞形态不规则,只是松散地覆盖卵母细胞。综上所述,这些观察结果表明ceh-18是鞘细胞分化的关键决定因素,这是正常减数分裂成熟和排卵所必需的功能。