| 摘要: |
| 【目的】克隆牦牛（Bos grunniens）水通道蛋白11（Aquaporin-11, AQP11)的基因序列，研究AQP11在不同发育阶段牦牛睾丸组织中的表达水平。【方法】采集初生（出生3 d）、青年（1岁）、成年（4岁）和老年（8岁）牦牛睾丸样品，通过RT-PCR技术扩增得到AQP11 基因的cDNA全长序列，对其进行生物信息学分析。采用qRT-PCR、Western blot方法对不同发育阶段牦牛睾丸中AQP11 基因及蛋白的表达水平进行检测，采用免疫组织化学法对AQP11在不同发育阶段牦牛睾丸中的表达进行定位。【结果】成功克隆了牦牛AQP11 基因cDNA全长序列，其长度为460 bp，共编码144个氨基酸。AQP11分子式:C710H1135N187O191S12，为稳定性蛋白，保守性较高。qRT-PCR结果显示，随着年龄的增长，AQP11 基因在牦牛睾丸中的表达水平呈先上升后略下降并趋于平稳的趋势，在青年牦牛睾丸中的表达水平显著高于其他年龄组（P＜0.05），其他年龄组间差异不显著（P＞0.05）。Western blot结果显示，随着牦牛年龄的增长，AQP11蛋白表达水平也呈先上升后略下降并趋于平稳的趋势，在青年牦牛睾丸中的表达水平最高，在初生牦牛睾丸中的表达水平显著低于其他年龄组（P＜0.05）。免疫组织化学试验结果显示，AQP11蛋白在睾丸中主要表达于精子、精子细胞、间质细胞、支持细胞和管周肌样细胞，各年龄段表达位置基本一致。【结论】 克隆了牦牛AQP11 基因cDNA全长序列，明确了其生物信息学特征。AQP11 基因在青年牦牛睾丸中的表达水平显著高于其他年龄组（P＜0.05），AQP11蛋白在青年牦牛中的表达水平显著高于初生牦牛（P＜0.05）。推测AQP11在牦牛精子发育、成熟及膜的完整性方面发挥着重要作用。 |
| 关键词: 牦牛 睾丸 水通道蛋白11基因 繁殖能力 高寒牧区 |
| DOI：10.13207/j.cnki.jnwafu.2025.05.001 |
| 分类号: |
| 基金项目:国家自然科学基金项目(31972760)；甘肃省教育厅产业支撑引导项目(2019C-03) |
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| Clonal analysis of the AQP11 gene in yak and its expression in the testis at different developmental stages |
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LIU Minqing1, CHEN Wendong1, HE Yupeng1, SU Manchun1, PAN Yangyang2, CUI Yan2, YU Sijiu2
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1.Technique College of Agriculture and Forestry,Longnan Normal University,Chengxian,Gansu 742500,China;2.Gansu Province Livestock Embryo Engineering Research Center,College of Veterinary Medicine,Gansu Agricultural University, Lanzhou,Gansu 730070,China
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| Abstract: |
| 【Objective】The gene sequence of aquaporin-11 (AQP11) of yak (Bos grunniens) was cloned to explore the expression level of AQP11 in yak testicular tissues at different developmental stages.【Method】 Testicular samples were collected from newborn (3 days after birth),young (1 year old),adult (4 years old) and old (8 years old) yaks.The full length cDNA sequence of AQP11 gene was amplified by RT-PCR and analyzed by bioinformatics.The expression of AQP11 gene and protein in yak testis at different developmental stages was detected by qRT-PCR and Western blot.The expression of AQP11 in yak testis at different developmental stages was localized by immunohistochemistry.【Result】 The full-length cDNA sequence of yak AQP11 gene was successfully cloned,which was 460 bp in length and encoded 144 amino acids.AQP11,whose molecular formula was C710H1135N187O191S12,was a stable and highly conserved protein.The results of qRT-PCR showed that with the increase of age,the expression level of AQP11 gene in yak increased firstly,then decreased slightly and finally tended to be stable.The expression level of AQP11 gene in young yak testis was significantly higher than that in other age groups (P＜0.05).There was no significant difference among other age groups (P＞0.05).Western blot results showed that with the increase of yak age,the expression level of AQP11 protein also increased first and then decreased slightly and tended to be stable,reaching the highest in the testis of young yak,and the expression level in the testis of newborn yak was significantly lower than that of other age groups (P＜0.05).The results of Immunohistochemistry showed that AQP11 protein was mainly expressed in sperm,sperm cells,leydig cells,supporting cells and peritubular myoid cells in the testis,and the expression positions were basically the same in all age groups.【Conclusion】The full-length cDNA sequence of yak AQP11 gene was cloned and its bioinformatic characteristics were clarified.The expression of AQP11 gene in young yak testis was significantly higher than that in other age groups (P＜0.05),and the expression level of AQP11 protein in young yak was significantly higher than that in newborn yak (P＜0.05).It is speculated that AQP11 plays an important role in sperm development,maturation and membrane integrity in yak testis. |
| Key words: yak testis AQP11 gene reproductive capacity alpine pastoral area |
