Objective:To explore the effect of Cfap65 deficiency on mouse spermatogenesis. Methods:CRISPR/Cas9 technology was utilized to construct Cfap65 deficient mice. PCR and Sanger sequencing were adopted to identify mouse genotypes. Mice were divided into Cfap65-/- group ( n=3) and wild-type (WT) group ( n=3) based on genotypes of mice. Fertility test was applied to evaluate the fertility of mice. Sperm morphology of Cfap65 deficient mice was observed by hematoxylin-eosin (HE) staining, immunofluorescence and transmission electron microscope. Real-time fluorescent quantitative PCR was used to detect the expression of Cfap65 mRNA in heart, liver, spleen, lung, kidney and testis tissues of mice. Results:Cfap65 deficient male mice were completely infertile. Compared with wild-type male mice, Cfap65 deficient mice had fewer and less motile epididymal spermatozoa, whose flagellums tend to be short, curled, bent and even absent, and heads tend to be deformed (1.67%±0.44% vs. 33.00%±1.53%), and the differences were statistically significant ( P<0.001). Besides, Cfap65 deficiency led to anomalous structure of manchette of mice. Cfap65 was highly expressed in the testes and lung of adult mice, and the expression of Cfap65 in testes embodied a sharp increase trend from mice aged 4 to 6 weeks (901.90±33.19 vs. 2 144.00±22.92), and the differences were statistically significant ( P<0.001). Conclusion:The expression of Cfap65 is tissue-specific, and the deletion of Cfap65 leads to spermatogenesis failure in male mice, which might be related to the dysfunction of intra-manchette transport.

Objective:To explore the effect of Cfap65 deficiency on mouse spermatogenesis. Methods:CRISPR/Cas9 technology was utilized to construct Cfap65 deficient mice. PCR and Sanger sequencing were adopted to identify mouse genotypes. Mice were divided into Cfap65-/- group ( n=3) and wild-type (WT) group ( n=3) based on genotypes of mice. Fertility test was applied to evaluate the fertility of mice. Sperm morphology of Cfap65 deficient mice was observed by hematoxylin-eosin (HE) staining, immunofluorescence and transmission electron microscope. Real-time fluorescent quantitative PCR was used to detect the expression of Cfap65 mRNA in heart, liver, spleen, lung, kidney and testis tissues of mice. Results:Cfap65 deficient male mice were completely infertile. Compared with wild-type male mice, Cfap65 deficient mice had fewer and less motile epididymal spermatozoa, whose flagellums tend to be short, curled, bent and even absent, and heads tend to be deformed (1.67%±0.44% vs. 33.00%±1.53%), and the differences were statistically significant ( P<0.001). Besides, Cfap65 deficiency led to anomalous structure of manchette of mice. Cfap65 was highly expressed in the testes and lung of adult mice, and the expression of Cfap65 in testes embodied a sharp increase trend from mice aged 4 to 6 weeks (901.90±33.19 vs. 2 144.00±22.92), and the differences were statistically significant ( P<0.001). Conclusion:The expression of Cfap65 is tissue-specific, and the deletion of Cfap65 leads to spermatogenesis failure in male mice, which might be related to the dysfunction of intra-manchette transport.