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细胞周期蛋白 A 在日本沼虾卵巢发育中的功能分析和分子特征。

Function analysis and molecular characterization of cyclin A in ovary development of oriental river prawn, Macrobrachium nipponense.

机构信息

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China.

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, Jiangsu, China.

出版信息

Gene. 2021 Jul 1;788:145583. doi: 10.1016/j.gene.2021.145583. Epub 2021 Mar 20.

DOI:10.1016/j.gene.2021.145583
PMID:33753150
Abstract

Macrobrachium nipponense has the characteristics of fast ovarian development cycle, which leads to the coexistence of multiple generations, the reduction of commodity specifications and the low economic benefit. Therefore, the study on the mechanism of ovarian development is of great significance to the development of industry. Cyclin A (CycA)is a key gene regulating ovarian development in vertebrates, but little information was available for its function in crustaceans. In this study, the full-length cDNA of Mn-CycA was obtained from the ovary. The full-length cDNA (2033 bp) with an open reading frame of 1368 bp, encoded a 456-amino acid protein. qRT-PCR revealed tissue-specific expression pattern of Mn-CycA, with abundant expression in the ovary. Results in different developmental stages of ovary indicated that Mn-CycA expression is positively correlated with ovarian maturation. qRT-PCR In different developmental stages, the expression of Mn-CycA mRNA gradually increased during the embryonic stage and decreased significantly on the first day of the hatching stage. At the 25th day of the metamorphosis stage, the expression level of Mn-CycAmRNA in female shrimp was 3.5 times higher than that in male shrimp, which may be related to the proliferation of oogonia and the formation of oocytes. In situ hybridization (ISH) of ovary showed Mn-CycA was examined in all stages and was mainly located in oogonia and oocytes. Compared with the control group, the obvious change of gonad somatic index (GSI) proved that injection of Mn-CycA dsRNA could delay the ovarian development cycle, which provided strong evidence for the involvement of Mn-CycA in ovarian maturation and oogenesis, and expanded a new perspective for studying the fast ovarian development cycle in M. nipponense.

摘要

日本沼虾具有卵巢发育周期快的特点,这导致了多个世代的共存、商品规格的减少和低经济效益。因此,研究卵巢发育的机制对产业发展具有重要意义。细胞周期蛋白 A(CycA)是调节脊椎动物卵巢发育的关键基因,但甲壳动物中关于其功能的信息很少。本研究从卵巢中获得了 Mn-CycA 的全长 cDNA。全长 cDNA(2033bp)的开放阅读框为 1368bp,编码 456 个氨基酸的蛋白质。qRT-PCR 显示 Mn-CycA 在组织中具有特异性表达模式,在卵巢中表达丰富。卵巢不同发育阶段的结果表明,Mn-CycA 的表达与卵巢成熟呈正相关。qRT-PCR 在不同的发育阶段,Mn-CycA mRNA 的表达在胚胎期逐渐增加,并在孵化期的第一天显著下降。在变态发育阶段的第 25 天,雌虾 Mn-CycAmRNA 的表达水平比雄虾高 3.5 倍,这可能与卵原细胞的增殖和卵母细胞的形成有关。卵巢原位杂交(ISH)显示 Mn-CycA 在所有阶段都有检测到,主要位于卵原细胞和卵母细胞中。与对照组相比,性腺体指数(GSI)的明显变化证明了 Mn-CycA dsRNA 的注射可以延缓卵巢发育周期,这为 Mn-CycA 参与卵巢成熟和卵子发生提供了有力证据,并为研究日本沼虾快速卵巢发育周期提供了新的视角。

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