The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, 361021, China.
Mol Biol Rep. 2012 Apr;39(4):4327-35. doi: 10.1007/s11033-011-1220-5. Epub 2011 Aug 13.
VASA is one of the important regulatory factors that determine the development of the reproductive system. However, no information on vasa gene from Pleocyemata Brachyura is available. By using Race, we obtained a full-length cDNA of Sp-vasa of the green mud crab Scylla paramamosain. The full-length (2,851 bp) cDNA of vasa encodes a peptide of 631 amino acids. Real-time PCR results indicated that the expression level of Sp-vasa in the growth stage of ovary was higher than in the maturation stage, and in stage I and II of testis, the expression level of Sp-vasa were higher than in stage III. By using in situ hybridization, Sp-vasa RNAs were detected in the large part of oocyte plasm in stage I, nucleus zone in stage III and perinuclear zone in stage V. As the size of oocytes increases during oogenesis, the signals change from strong to weak. In addition, in stage I and II of testis, the expression levels of Sp-vasa were higher than in stage III, and the hybridization intensity of Sp-vasa gene gradually increased during spermatogenesis from spermatogonia to spermatids. However, no hybridization signal was detected in spermatozoon. Real-time PCR and in situ hybridization were consistent. These findings suggest that Sp-vasa is likely to serve as a useful and specific marker for germ cell development of S. paramamosain.
VASA 是决定生殖系统发育的重要调节因子之一。然而，尚无关于十足目短尾亚目的 vasa 基因的信息。通过使用 Race，我们获得了青蟹 Scylla paramamosain 的 Sp-vasa 的全长 cDNA。全长（2851bp）的 vasa 编码一个 631 个氨基酸的肽。实时 PCR 结果表明，Sp-vasa 在卵巢生长阶段的表达水平高于成熟阶段，在睾丸的 I 期和 II 期，Sp-vasa 的表达水平高于 III 期。通过原位杂交，在 I 期的卵母细胞质的大部分、III 期的核区和 V 期的核周区检测到 Sp-vasa RNA。随着卵母细胞发生过程中卵母细胞的增大，信号从强变弱。此外，在睾丸的 I 期和 II 期，Sp-vasa 的表达水平高于 III 期，并且 Sp-vasa 基因的杂交强度在从精原细胞到精细胞的精子发生过程中逐渐增加。然而，在精子中未检测到杂交信号。实时 PCR 和原位杂交结果一致。这些发现表明，Sp-vasa 可能是 S. paramamosain 生殖细胞发育的有用且特异性标记物。
