罗氏沼虾Doublesex基因的cDNA克隆及性别二态性表达分析

cDNA cloning and sex dimorphism expression analysis of the Doublesex gene in the prawn Macrobrachium rosenbergii

摘要:

克隆了罗氏沼虾(Macrobrachium rosenbergii)含DM结构域的Dsx基因(MroDsx)部分序列,DM结构域包含5个保守的半胱氨酸(Cysteine)和2个组氨酸(Histidine), 且与中国对虾(Fenneropenaeus chinensis) Dsx基因DM结构域具有73%的相似性。荧光定量PCR(Quantitative real time PCR,qPCR)结果显示:MroDsx基因仅在性腺中特异性表达,且在精巢中的表达量显著高于卵巢;在精巢发育早期(主要是精原细胞)高表达,随着精巢的发育MroDsx基因的表达量逐渐降低。原位杂交(In situ hybridization,ISH)结果进一步显示:MroDsx转录本在精原细胞表达量高,在精母细胞以及精细胞中较弱,在成熟的精子中不表达,与荧光定量的结果一致。基于对MroDsx转录本的表达分析,推测MroDsx可能参与罗氏沼虾精巢的发育过程。

Abstract:

In this study, we cloned a partial cDNA sequence of Dsx (MroDsx) containing DM domain in the fresh water prawn Macrobrachium rosenbergii. The DM domain includes five conserved cysteines and two histidine, and shares 73% similarity with those of Dsx gene of the Chinese shrimp Fenneropenaeus chinensis. Quantitative PCR (qPCR) showed that MroDsx was specifically expressed in gonads, and the expression level in testis was significantly higher than that in ovary. The expression of MroDsx is particularly high in the early testis, and gradually decreased with the development of testis. In situ hybridization (ISH) results further showed MroDsx transcript is highly expressed in spermatogonium, weak in the spermatocyte and spermatid, and not expressed in spermatozoa, which is highly consistent with the results of qPCR. Based on the expression analysis of MroDsx transcript, it is implied that MroDsx may be involved in the testis development of Macrobrachium rosenbergii.