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目的:探讨一种新的睾丸特异性蛋白FSCB(Fibrous sheath CABYR binding protein)在精子内的精细动态表达和迁移过程,并研究该蛋白在正常和畸形精子中的形态定位特征。方法:应用过碘酸-希夫氏染色获得鼠睾丸曲精小管上皮中精子发生的时相性背景特征,然后应用间接免疫荧光方法对FSCB蛋白在睾丸和精子内的动态表达和迁移过程进行观察。结果:FSCB最初表达于长精细胞的第11步(时相Ⅺ),呈胞浆内弥漫性染色;随之从第11步(时相Ⅺ)至第15步(时相Ⅵ)表达逐渐加强;在精子形成的第15步(时相Ⅳ至时相Ⅵ)FSCB开始向精子鞭毛迁移,最终于第16步(时相Ⅷ)FSCB蛋白整体局限定位于睾丸精子鞭毛上,胞浆内荧光染色消失。进一步研究显示该蛋白在正常精子中定位于鞭毛主段、精子头部、鞭毛颈段、中段和末段均无表达。在畸形精子中,该蛋白表达存在3种异常改变:①鞭毛内无阳性表达;②表达节段前移并缩短,提示这类精子缺乏中段;③呈现间断分布的串珠状改变。结论:FSCB是纤维鞘的重要组成蛋白,在精子纤维鞘形成的后期装配于其表面,可能是纤维鞘生物合成的最后装配步骤。该蛋白在畸形精子中存在明显表达异常,其表达异常与精子运动功能之间的关系有待深入研究。
OBJECTIVE: To investigate the fine dynamic expression and migration of a novel testicular-specific protein FSCB in sperm and to investigate the morphological localization of this protein in normal and abnormal sperm. Methods: The temporal characteristics of spermatogenesis in testicular seminiferous tubule epithelial cells were obtained by periodic acid-Schiff staining. The dynamic expression and migration of FSCB protein in testis and sperm were observed by indirect immunofluorescence staining. RESULTS: FSCB was initially expressed in the cytoplasm in 11th stage (phase Ⅺ) of spermatogenic cells, and gradually increased from 11th stage (phase Ⅺ) to 15th stage (phase Ⅵ) ; FSCB begins to migrate to sperm flagella in the 15th stage of spermatogenesis (phase Ⅳ to phase Ⅵ), and finally in step 16 (phase Ⅷ), the FSCB protein is localized localized to testicular sperm flagella. Intracytoplasmic fluorescence staining disappear. Further studies showed that the protein was located in the main segment of flagella in normal sperm, sperm head, flagella neck segment, no expression in the middle and distal segments. In abnormal sperm, there are three abnormal changes in the expression of the protein: ① no expression within the flagellum; ② forward and shorten the expression segment, suggesting that these sperm lack of middle; ③ showed intermittent beaded changes. CONCLUSIONS: FSCB is an important constituent of fiber sheath, which is assembled on the surface of sperm sheath in the later stage of sperm sheath formation, which may be the final assembly step of fiber sheath biosynthesis. The abnormal expression of the protein in abnormal sperm abnormalities, the relationship between abnormal expression and sperm motility needs further study.