动物造模 z-bio 2022-05-07 436

　　Objective To study the transcript levels of vitamin D metabolizing enzymes and vitamin D receptors in the testis of adult tree shrew, detect the protein expression of CYP24A1, and explore the causes of reproductive disorders in male experimental tree shrew.

　　Methods Male tree shrew aged 2-3 years were selected and divided into a normal group with a successful reproductive history and a reproductive disorder group without a reproductive history according to individual reproductive profiles, with 5 in each group. One testis was surgically removed, and histopathological changes were observed by pathology; real-time fluorescence quantitative PCR was used to detect the changes of vitamin D receptor and vitamin D metabolizing enzyme mRNA levels in testis tissue, Western blotting was used to detect the protein expression of CYP24A1, and at the same time The expression of sperm CYP24A1 was observed by immunofluorescence.

　　Results Compared with the reproductive disorder group, the seminiferous tubules were more closely arranged and the spermatogenic cells were more in the normal group. Compared with the normal group, the mRNA levels of VDR, CYP2R1 and CYP27A1 in the testis tissue of tree shrew in the reproductive disorder group had no significant difference (P>0.05), while the mRNA expression and protein levels of CYP24A1 were significantly lower than those in the normal group (P<0.05). .05). At the same time, the results of immunofluorescence showed that the spermatozoa derived from the testis in the normal group had high expression of CYP24A1 protein in the head, neck and midsection, while the reproductive disorder group showed low expression or no expression.

　　Conclusion CYP24A1 is directly related to the successful fertility of male tree shrew, suggesting that CYP24A1 can be used as an important indicator to judge the fertility of male tree shrew.