Oka T, Futai M
Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Japan.
J Biol Chem. 2000 Sep 22;275(38):29556-61. doi: 10.1074/jbc.M002756200.
Immunofluorescence analysis indicated that VHA-11, the C subunit of Caenorhabditis elegans V-ATPase, was localized in dot-like structures around the nuclei of early embryonic cells and was also detected in embryonic intestinal cells after comma stage. Vital staining with acridine orange showed that the intestinal cells had acidic compartments generated by V-ATPase, consistent with the intracellular localization of VHA-11. RNA interference could efficiently silence vha-11 gene expression: introduction of vha-11 double strand RNA led to embryonic lethality. Worms injected with the vha-11 double strand RNA produced embryos that became lethal. The development of embryos was arrested at various stages. However, their numbers gradually decreased, and the worms eventually became sterile due to the failure of ovulation. Similar results were obtained for RNA interference of the V-ATPase proteolipid genes. These results suggest that V-ATPases, and thus inside-acidic organelles, are required for ovulation and embryogenesis.
免疫荧光分析表明，秀丽隐杆线虫V-ATP酶的C亚基VHA-11定位于早期胚胎细胞核周围的点状结构中，在逗号期后的胚胎肠道细胞中也能检测到。吖啶橙活体染色显示，肠道细胞具有由V-ATP酶产生的酸性区室，这与VHA-11的细胞内定位一致。RNA干扰能够有效沉默vha-11基因表达：导入vha-11双链RNA会导致胚胎致死。注射了vha-11双链RNA的线虫所产生的胚胎会致死。胚胎发育在各个阶段停滞。然而，胚胎数量逐渐减少，线虫最终因排卵失败而不育。对V-ATP酶蛋白脂质基因进行RNA干扰也得到了类似结果。这些结果表明，V-ATP酶以及由此产生的细胞内酸性细胞器是排卵和胚胎发生所必需的。
