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目的利用fscb基因敲除小鼠,探讨FSCB蛋白在小鼠精子运动和授精功能中的作用。方法应用免疫荧光观察fscb基因敲除雄鼠睾丸组织及精子中FSCB蛋白表达情况;应用计算机辅助精液分析(computer assisted sperm analysis,CASA)系统分析fscb基因敲除小鼠精子在体外的运动特征、活率和运动参数改变情况;在体外获能培养条件下检测精子的体外受精能力;将fscb基因敲除纯合子(KO)、杂合子(HET)、野生型(WT)雄鼠分别与野生型C57BL/6J成年雌鼠交配,观察雌鼠怀孕率、怀孕胚胎数量、自然产仔数及子代成年雄鼠生殖腺形态学是否存在差异。结果免疫荧光显示纯合子雄鼠睾丸组织及精子中均无FSCB蛋白表达,杂合子雄鼠睾丸组织及精子中FSCB蛋白表达与野生型相比明显减低。CASA显示各组雄鼠精子数、活率、各项运动参数差异均无统计学意义(P>0.05);野生型、杂合子和纯合子雄鼠精子与透明带完整卵细胞及与无透明带卵细胞结合的精子平均数、卵细胞的受精率差异均无统计学意义(P>0.05);各组雄鼠对应雌鼠的受孕率差异无统计学意义(P>0.05);野生型、杂合子和纯合子对应雌鼠受孕后每只雌鼠胚胎数分别为(7.00±1.41)、(7.33±1.53)、(7.20±1.48)只,每窝自然产仔数分别为(6.00±2.00)、(6.75±1.71)、(7.80±1.30)只,组间比较差异无统计学意义(P>0.05);各组子代成年雄鼠生殖腺发育差异无统计学意义(P>0.05)。结论敲除fscb基因对雄性小鼠精子的运动功能及体内外受精能力均无明显影响,提示该基因所编码的FSCB蛋白的功能对于精子的运动和授精能力的维持可能是非必需的或可被替代的。
Objective To investigate the role of FSCB protein in sperm motility and insemination in mice using fscb knockout mice. Methods The expression of FSCB protein in testis tissue and sperm of fscb knockout male rats was observed by immunofluorescence. The movement characteristics of sperms in fscb knockout mice were analyzed by computer assisted sperm analysis (CASA) Rate, and exercise parameters were measured. In vitro fertilization was used to test sperm in vitro fertilization ability. Fscb knockout homozygotes (KO), heterozygotes (HET) and wild type (WT) males were compared with wild type C57BL / 6J adult female mice were mating, observing the female pregnancy rate, the number of pregnant embryos, the number of natural littermates and male offspring of adult male gonadal morphology is there any difference. Results Immunofluorescence showed no expression of FSCB protein in testis tissue and sperm of homozygous male rats, and the expression of FSCB protein in testis tissue and sperm of heterozygous male rats was significantly lower than that of wild type. CASA showed no significant differences in the number of sperm, viability and motor parameters between males in each group (P> 0.05). In the spermatozoa and zona pellucida of wild type, heterozygous and homozygous males, There were no significant differences in the fertilization rate of oocytes between the two groups (P> 0.05). There was no significant difference in the fertility rate between the male and female mice (P> 0.05). The wild type, heterozygote and pure The number of embryos of each female mouse after conception was (7.00 ± 1.41), (7.33 ± 1.53) and (7.20 ± 1.48), respectively. The number of natural litter size in each litter was (6.00 ± 2.00) and (6.75 ± 1.71, and 7.80 ± 1.30, respectively. There was no significant difference between the two groups (P> 0.05). There was no significant difference in the development of gonadal between adult male and female rats in each group (P> 0.05). Conclusion The deletion of fscb gene had no significant effect on sperm motor function and in vitro and in vivo fertilization ability of male mice, suggesting that the function of FSCB protein encoded by this gene may not be necessary or may be substituted for the maintenance of sperm motility and fertilization ability of.