Takamura Katsumi, Fujimura Miyuki, Yamaguchi Yasunori
Department of Marine Biotechnology, Faculty of Engineering, Fukuyama University, 729-0292, Japan.
Dev Genes Evol. 2002 Feb;212(1):11-8. doi: 10.1007/s00427-001-0204-1. Epub 2002 Jan 24.
The origin of germ cells in the ascidian is still unknown. Previously, we cloned a vasa homologue (CiVH) of Ciona intestinalis from the cDNA library of ovarian tissue by polymerase chain reaction and showed that its expression was specific to germ cells in adult and juvenile gonads. In the present study, we prepared a monoclonal antibody against CiVH protein and traced the staining for this antibody from the middle tailbud stage to young adulthood. Results showed that positive cells are present in the endodermal strand in middle tailbud embryos and larvae. When the larval tail was absorbed into the trunk during metamorphosis, the CiVH-positive cells migrated from the debris of the tail into the developing gonad rudiment, and appeared to give rise to a primordial germ cell (PGC) in the young juvenile. The testis rudiment separated from the gonad rudiment, the remainder of which differentiated into the ovary. PGCs of the testis rudiment and the ovary rudiment differentiated into spermatogenic and oogenic cells, respectively. When the larval tail containing the antibody-positive cells was removed, the juveniles did not contain any CiVH-positive cells after metamorphosis, indicating that the PGCs in the juvenile originated from part of the larval tail. However, even in such juveniles, positive cells newly appeared in the gonad rudiment at a later stage. This observation suggests that a compensatory mechanism regulates germline formation in C. intestinalis.
海鞘生殖细胞的起源仍然未知。此前,我们通过聚合酶链反应从卵巢组织的cDNA文库中克隆了肠鳃纲动物的一个vasa同源物(CiVH),并表明其表达在成年和幼年性腺的生殖细胞中具有特异性。在本研究中,我们制备了一种针对CiVH蛋白的单克隆抗体,并追踪了该抗体从尾芽中期到成年早期的染色情况。结果显示,在尾芽中期胚胎和幼虫的内胚层索中存在阳性细胞。在变态过程中,当幼虫尾巴被吸收到躯干中时,CiVH阳性细胞从尾巴的残骸迁移到发育中的性腺原基,并似乎在幼年早期产生了原始生殖细胞(PGC)。睾丸原基与性腺原基分离,性腺原基的其余部分分化为卵巢。睾丸原基和卵巢原基的PGC分别分化为生精细胞和生卵细胞。当去除含有抗体阳性细胞的幼虫尾巴时,变态后的幼体不含任何CiVH阳性细胞,这表明幼体中的PGC起源于幼虫尾巴的一部分。然而,即使在这些幼体中,后期性腺原基中也会新出现阳性细胞。这一观察结果表明,一种补偿机制调节着肠鳃纲动物的种系形成。