Huang Xiaoshuai, Ye Haihui, Huang Huiyang, Yang Yanan, Gong Jie
College of Ocean and Earth Sciences, Xiamen University, Xiamen 361005, China.
College of Ocean and Earth Sciences, Xiamen University, Xiamen 361005, China.
Gen Comp Endocrinol. 2014 Aug 1;204:229-38. doi: 10.1016/j.ygcen.2014.06.002. Epub 2014 Jun 12.
Insulin-like androgenic gland hormone (IAG) produced by androgenic gland (AG) in male crustaceans is regarded as a key regulator of sex differentiation. As a member of the insulin/insulin-like growth factor family, IAG is also likely involved in regulating somatic growth. In this study, a full-length cDNA of IAG (termed Sp-IAG) was isolated from the mud crab, Scylla paramamosain. Genomic DNA of Sp-IAG was also cloned, analysis of which reveals that Sp-IAG gene is organized in a 4 exon/3 intron manner. RNA in situ hybridization analysis detected positive signals in both type I and type II AG cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that Sp-IAG was expressed not only in AG, but also in many other tissues. Sp-IAG expression levels in ovaries were examined at different stages of ovarian development (stages I to V); it was found that the expression was maintained at low levels during undeveloped stage (stage I) to late vitellogenic stage (stage IV) and then increased significantly at mature stage (stage V), suggesting that Sp-IAG may participate in inhibiting oocyte growth and vitellogenesis. The expression pattern of Sp-IAG during the molting cycle of the first stage crabs (C1) was also determined. Sp-IAG expression level continuously decreased from 0 h C1 (postmolt) crabs to 96 h C1 (premolt) crabs, and then increased significantly in the newly molted second stage crabs (C2, postmolt). The combined results suggested for the first time that IAG is involved in regulating ovarian development and somatic growth in crustaceans.
雄性甲壳类动物的雄激素腺(AG)产生的胰岛素样雄激素腺激素(IAG)被认为是性别分化的关键调节因子。作为胰岛素/胰岛素样生长因子家族的成员,IAG也可能参与调节体细胞生长。在本研究中,从拟穴青蟹(Scylla paramamosain)中分离出IAG的全长cDNA(命名为Sp-IAG)。还克隆了Sp-IAG的基因组DNA,分析表明Sp-IAG基因以4个外显子/3个内含子的方式组织。RNA原位杂交分析在I型和II型AG细胞中均检测到阳性信号。逆转录-聚合酶链反应(RT-PCR)分析表明,Sp-IAG不仅在AG中表达,还在许多其他组织中表达。检测了卵巢发育不同阶段(I至V期)卵巢中Sp-IAG的表达水平;发现其表达在未发育阶段(I期)至卵黄发生后期(IV期)维持在低水平,然后在成熟阶段(V期)显著增加,表明Sp-IAG可能参与抑制卵母细胞生长和卵黄发生。还确定了第一期蟹(C1)蜕皮周期中Sp-IAG的表达模式。Sp-IAG表达水平从C1期0小时(蜕皮后)蟹持续下降至C1期96小时(蜕皮前)蟹,然后在新蜕皮的第二期蟹(C2,蜕皮后)中显著增加。综合结果首次表明IAG参与调节甲壳类动物的卵巢发育和体细胞生长。