摘要
用原位杂交技术,以地高辛标记的反义RNA为探针,检测了金鱼(Carassiusauratus)DEAD box家族基因vasa在卵子及精子发生中的分布及表达。结果表明在金鱼卵子发生中,在各个时期的卵母细胞的胞质中均有金鱼vasaRNA的杂交信号表达。在Ⅰ、Ⅱ期卵母细胞中vasaRNA的杂交信号强烈,均匀地分布在整个胞质。随着卵母细胞的生长发育及卵黄的积累,Ⅲ、Ⅳ期卵母细胞胞质中vasaRNA的杂交信号急剧减弱,而外周皮层区域,其阳性信号仍较强。在金鱼精子发生中,在精原细胞和初级精母细胞中可检测到金鱼vasaRNA杂交信号,后者的阳性信号比前者微弱;而精子细胞中没有阳性信号。推测vasa基因在金鱼精子发生早期发挥着重要作用;而在卵子发生中主要作为遗传信息储备物质,用于调控早期胚胎发育过程中原始生殖细胞的形成与分化。
Using in situ hybridization with DIG antisense RNA probe, we detected the expression and distributing of goldfish (Carassius auratus) DEAD-box family gene vasa during oogenesis and spermatogenesis. The results showed that during oogenesis, the positive signals of vasa RNA were detected in the cytoplasm of oocytes at all stages. In stage Ⅰand Ⅱ oocytes, strong hybridization signals of vasa RNA were dispersed uniformly throughout the cytoplasm. With the development of oocytes and accumulation of yolk, the signals of vasa RNA significantly decreased in most of the regions while remained strong at cortical region in stage Ⅲ, Ⅳ oocytes. During spermatogenesis, the positive signals of vasa RNA could only be detected in spermatogenia and primary spermatocytes while the signals in the former were much stronger than in the latter. However, no signals could be detected in spermatids. Our results suggested that gene vasa may play an important role in the early stage of goldfish spermatogenesis; vasa RNA is deposited in oocytes and subsequently regulate the germ cells specification and differentiation during the early embryogenesis.
基金
国家"863"项目(2001AA222071)
国家自然科学基金资助项目(30270675)
