陈康轩^1,2, 李诗豪¹, 李富花¹
1.中国科学院海洋研究所实验海洋生物学重点实验室, 山东 青岛 266071;2.中国科学院大学北京 100049

摘要:
位于甲壳动物眼柄的神经内分泌器官在调控甲壳动物的繁殖过程中发挥着重要作用。已有证据表明“眼柄-促雄性腺-精巢”内分泌轴调控十足目雄性甲壳动物精巢的发育，但是该过程的分子机制仍不清楚。本研究以凡纳滨对虾（Litopenaeus vannamei）为实验对象，研究了切除雄性对虾的单侧眼柄对促雄性腺内胰岛素样促雄性腺素基因（LvIAG）和精巢内基因表达的影响。结果表明，切除单侧眼柄后，LvIAG基因的表达明显上调；比较了眼柄切除前后对虾精巢的转录组数据，共有267个基因的表达量出现明显变化。对精巢内差异表达基因进行功能分析发现，多个参与性腺发育和内分泌调控的基因发生表达上调，如Doublesex（Dsx）、保幼激素环氧水解酶（juvenile hormone epoxide hydrolase，JHEH）、细胞色素P450酶系基因以及泛素化系统相关基因等。对差异表达基因进行荧光定量PCR验证，结果与转录组结果一致，证实了转录组结果的可靠性。研究结果表明，对虾眼柄调控精巢的发育很可能是通过影响促雄性腺中LvIAG的表达，进而调控精巢内Dsx、JHEH、内分泌及泛素化系统相关基因的表达实现的。研究结果对深入理解甲壳动物精巢发育的分子调控机制提供了重要依据，对甲壳动物的人工繁育也具有重要指导意义。
关键词:对虾眼柄胰岛素样促雄性腺素精巢发育
DOI：10.11759/hykx20210511001
分类号:S917.4
基金项目:国家重点研发计划项目（2018YFD0900202）；中以（NSFC-ISF）国际（地区）合作与交流项目（31861143047）

Regulatory mechanisms of the eyestalk-androgenic gland-testis endocrine axis on testis development in Litopenaeus vannamei
CHEN Kang-xuan^1,2, LI Shi-hao¹, LI Fu-hua¹
1.Key Laboratory of Experimental Marine Biology, Institute of Oceanology Chinese Academy of Sciences, Qingdao 266071, China;2.University of Chinese Academy of Sciences, Beijing 100049, China
Abstract:
The neuroendocrine system in crustacean eyestalks regulates the reproduction process. According to previous studies, the eyestalk-androgenic gland (AG)-testis endocrine axis regulates testis development in decapod crustacean species. However, the underlying molecular mechanism remains largely unknown. The Pacific white shrimp (Litopenaeus vannamei) was used in this study to investigate how unilateral eyestalk ablation regulates the expression of insulin-like AG hormones (LvIAG) in AG, and genes in the testis. According to the results, after unilateral eyestalk ablation, the LvIAG expression level was significantly upregulated. The transcriptome data of shrimp testis before and after unilateral eyestalk ablation were compared. A total of 267 genes were identified as differentially expressed genes (DEGS). Functional analysis showed that Some genes involved in gonad development and endocrine regulation, such as Doublesex (Dsx), juvenile hormone epoxide hydrolase (JHEH), genes related to the cytochrome P450 enzyme system, and genes related to the ubiquitylation system, were significantly upregulated. The expression of several DEGs were confirmed using quantitative real-time polymerase chain reaction (qRT-PCR), which revealed that their expression trends were consistent with those in the transcriptome data, implying that the transcriptome data were reliable. The results indicated that the endocrine system in eyestalk influenced testis development by regulating the expression of LvIAG in AG, which in turn regulated the expression of Dsx,JHEH, and genes in the endocrine and ubiquitylation systems in the testis. The results not only contribute to a better understanding of the molecular mechanisms regulating testis development but also provide significant direction for the artificial reproduction of crustaceans.
Key words:penaeid shrimpeyestalkinsulin-like androgenic gland hormonetestis development





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