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

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...

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Bibliographic Details
Published in:Animals (Basel) 2021-12, Vol.11 (12), p.3582
Main Authors: Xiang, Qiumeng, Wei, Chaoguang, Gao, Xinming, Chen, Yiner, Tang, Daojun, Zhu, Junquan, Hou, Congcong
Format: Article
Language:English
Subjects:
Alkaline phosphatase
Antioxidants
Apoptosis
Bcl-2 protein
Breeding
Caspase-3
Cell division
cell dysfunction
Chains
Cloning
Crustaceans
Dynein
dynein heavy chain
Enzymatic activity
Enzyme activity
Fluorescence
Gene expression
Genes
Genomics
Hsp70 protein
Immunofluorescence
Inhibitors
Insects
Localization
Molecular motors
Nitric-oxide synthase
p53 Protein
Portunus trituberculatus
Protein transport
Proteins
Reproductive system
Shellfish
Sodium
Sperm
Spermatids
Spermatogenesis
Superoxide dismutase
Testes
testis
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Summary: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 .
ISSN:2076-2615
2076-2615
DOI:10.3390/ani11123582