| 摘要: |
| 【目的】克隆分析绵羊联会复合中心组分蛋白1（synaptonemal complex central element protein 1，SYCE1）基因，检测其在雄性生殖轴系中的表达，探索SYCE1对雄性动物生殖发育的调控机制。【方法】以雄性绵羊（小尾寒羊）为研究对象，通过RT-PCR技术克隆SYCE1基因序列，并对该序列及其编码的蛋白进行生物信息学分析；利用qRT-PCR、Western blotting技术分析SYCE1在绵羊下丘脑-垂体-睾丸生殖轴及不同发育阶段（40日龄、3月龄和12月龄）睾丸中的表达情况；采用免疫组织化学染色方法检测SYCE1蛋白在不同发育阶段睾丸组织中的分布情况。【结果】核苷酸序列分析表明,SYCE1基因CDS全长972 bp，编码274个氨基酸。进化树结果表明,绵羊SYCE1氨基酸序列与山羊、欧洲普通牛、野牛、白尾鹿等的氨基酸同源性高；不同物种之间同源性分析发现，SYCE1基因在进化中高度保守；qRT-PCR、Western blotting检测结果表明，SYCE1 mRNA及其蛋白在绵羊的下丘脑、垂体、睾丸及附睾组织中均有表达，其中睾丸组织表达量极显著高于其他组织（P<0.01），对于不同发育阶段的睾丸组织SYCE1表达水平随着绵羊年龄的增加逐渐升高；免疫组织化学染色结果显示，SYCE1蛋白在40日龄定位于睾丸间质细胞和少量精原细胞，在3月龄定位于睾丸初级精母细胞、精原细胞和间质细胞，在12月龄定位于睾丸初级精母细胞、次级精母细胞、精原细胞、间质细胞和支持细胞。【结论】SYCE1在雄性绵羊生殖轴和不同发育阶段睾丸中均有表达，随着绵羊年龄的增加SYCE1在睾丸组织中的表达水平逐渐上调，说明其与绵羊睾丸器官发育成熟度相关，推断其参与雄性绵羊生殖轴调控。 |
| 关键词: 联会复合中心组分蛋白1（SYCE1）基因 绵羊 生殖轴 睾丸 |
| DOI： |
| 分类号: |
| 基金项目:甘肃省教育厅高校协同创新团队项目(2018C-15)；2018年度甘肃省委组织部重点人才项目(041045)；兰州市科技局创新人才重点项目（2015-RC-7） |
|
| Cloning of SYCE1gene in sheep (Ovis aries) and its expression in male reproductive axis |
|
YANG Yang,LI Zongshuai,DONG Weitao,et al
|
| Abstract: |
| 【Objective】The synaptonemal complex central element protein 1 (SYCE1) gene of sheep was cloned,its expression in the male reproductive axis was detected,and the regulatory mechanism of SYCE1 on male animal reproduction was explored.【Method】Male sheep (small-tailed Han sheep) was selected,and the SYCE1 gene sequence was cloned by RT-PCR technology.The sequence and its encoded protein were analyzed by bioinformatics.The qRT-PCR and Western blotting technology were used to analyze SYCE1.Expression in thalamus-pituitary-testis reproductive axis and testes at different developmental stages (40-day-old,3-month-old,and 12-month-old) was determined and immunohistochemistry staining was used to detect the distribution of SYCE1 protein in testicular tissue at different developmental stages.【Result】Nucleotide sequence analysis showed that the gene CDS was 972 bp in length and encoded 274 amino acids.The phylogenetic tree results showed that the amino acid sequence of Ovis aries SYCE1 was close to the high evolutionary level of amino acid homology of Capra hircus,Bos taurus,Bison bison bison,and Odocoileus virginianus.The homology analysis between different species found that SYCE1 gene was highly conserved in evolution,and qRT-PCR and Western blotting results showed that SYCE1 mRNA and protein expressed in hypothalamus,pituitary,testis and epididymis of sheep.The expression in testis was significantly higher than in other tissues (P<0.01).The testicular tissue SYCE1 expression level gradually increased with age.Immunohistochemistry staining results showed that SYCE1 protein was localized to testicular leydig cells and a few spermatogonia at the age of 40 days,localized in testicular spermatogonia,primary spermatocytes and leydig cells at the age of 3 months,and localized to testicular primary spermatocytes,secondary spermatocytes,spermatogonia,leydig cells and sertoli cells at the age of 12 months.【Conclusion】SYCE1 was expressed in reproductive axis and testes of male sheep at different developmental stages.As the age of sheep increased,SYCE1 expression level gradually increased in testicular tissues,indicating that it is related to the developmental maturity of testicular organs.It is inferred that it participates in reproductive axis regulation of male sheep. |
| Key words: synaptonemal complex central element protein 1(SYCE1) gene sheep reproductive axis testis |
