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动力蛋白重链的分子克隆及其抑制对……睾丸功能的影响

Molecular Cloning of Dynein Heavy Chain and the Effect of Dynein Inhibition on the Testicular Function of .

作者信息

Xiang Qiumeng, Wei Chaoguang, Gao Xinming, Chen Yiner, Tang Daojun, Zhu Junquan, Hou Congcong

机构信息

Key Laboratory of Applied Marine Biotechnology of Ministry of Education, School of Marine Sciences, Ningbo University, Ningbo 315832, China.

出版信息

Animals (Basel). 2021 Dec 17;11(12):3582. doi: 10.3390/ani11123582.

Abstract

Dynein is a motor protein with multiple transport functions. However, dynein's role in crustacean testis is still unknown. We cloned the full-length cDNA of cytoplasmic dynein heavy chain () gene and its structure was analyzed. Its expression level was highest in testis. We injected the dynein inhibitor sodium orthovanadate (SOV) into the crab. The distribution of dynein heavy chain (-DHC) in mature sperm was detected by immunofluorescence. The apoptosis of spermatids was detected using a TUNEL kit; gene expression in testis was detected by fluorescence quantitative PCR (qPCR). The expression of immune-related factors in the testis were detected by an enzyme activity kit. The results showed that the distribution of -DHC was abnormal after SOV injection, indicating that the function of dynein was successfully inhibited. Apoptosis-related genes and , and antioxidant stress genes and were significantly decreased, and anti-apoptosis gene - was significantly increased. The activities of superoxide dismutase (SOD) and alkaline phosphatase (AKP) were significantly decreased. The results showed that there was no apoptosis in testicular cells after dynein function was inhibited, but the cell function was disordered. This study laid a theoretical foundation for the further study of apoptosis in testis and the function of dynein in testis and breeding of .

摘要

动力蛋白是一种具有多种运输功能的马达蛋白。然而,动力蛋白在甲壳类动物睾丸中的作用仍不清楚。我们克隆了细胞质动力蛋白重链()基因的全长cDNA并分析了其结构。其在睾丸中的表达水平最高。我们将动力蛋白抑制剂原钒酸钠(SOV)注射到螃蟹体内。通过免疫荧光检测成熟精子中动力蛋白重链(-DHC)的分布。使用TUNEL试剂盒检测精子细胞的凋亡;通过荧光定量PCR(qPCR)检测睾丸中的基因表达。通过酶活性试剂盒检测睾丸中免疫相关因子的表达。结果表明,注射SOV后-DHC的分布异常,表明动力蛋白的功能被成功抑制。凋亡相关基因和以及抗氧化应激基因和显著降低,而抗凋亡基因-显著增加。超氧化物歧化酶(SOD)和碱性磷酸酶(AKP)的活性显著降低。结果表明,动力蛋白功能被抑制后睾丸细胞没有凋亡,但细胞功能紊乱。本研究为进一步研究睾丸凋亡以及动力蛋白在睾丸中的功能和的繁殖奠定了理论基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5890/8697902/07595813a1e6/animals-11-03582-g001.jpg

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