Biochemical Characteristics and Functional Studies of the Sperm-specific Calcium Channel CatSper in Tupaia belangeri
[Objective]The purpose of this experiment was to explore the biochemical characteristics of specific calcium channel CatSper and its subcellular location in Tupaia belangeri sperm,and to clarify the regulatory role of CatSper on the motility of Tupaia belangeri sperm.[Method]Sexually mature male Tupaia belangeri aged 1 to 2 years were selected,semen was collected after being killed with CO2,sperm motility was observed and protein was extracted.Western blotting and immunohistochemistry were used to detect the presence and subcellular localization of CatSper family in sperm.Different calcium channel inhibitors(CatSper inhibitor HC-056456,transient receptor potential vanillate subtype 1(TRPV1)inhibitor Capsazepine)co-incubated with progesterone(P4)were used to observe the motility of Tupaia belangeri sperm.Calcium probe method was used to detect the effects of different inhibitors and P4 co-incubation on calcium ion concentration of Tupaia belangeri sperm.[Result]CatSper and TRPV1 channel protein were present in Tupaia belangeri sperm,and CatSper was located in the sperm head and flagella.Compared with control group,sperm motility and calcium ion concentration were significantly decreased when HC-056456 concentrations were 100,200,400 and 1 000 nmol/L(P＜0.05),and Capsazepine concentrations were 10,20 and 40 μmol/L,sperm motility and calcium ion concentration were significantly decreased(P＜0.05).Compared with Capsazepine group,when HC-056456 was co-incubated with P4,the calcium ion concentration of Tupaia belangeri sperm was significantly increased,and there were significant differences between the two groups when P4 concentrations were 10,20 and 30 μmol/L(P＜0.05).When Capsazepine and HC-056456 were co-incubated with P4,there was no significant difference in sperm motility between the two groups(P＞0.05).[Conclusion]CatSper channel played an important role in the regulation of sperm motility and calcium ion transport in Tupaia belangeri,and the calcium ion concentration in sperm was increased through P4 activation.The results were of great significance for further research on the regulatory mechanism of sperm function and sperm fertilization ability.
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维普
