| 牦牛NGF基因克隆及其在生殖器官中的表达定位 |
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| 引用本文： | 邬建飞,刘宇,马瑶,蒋旭东,胡双阁,龚三你,字向东,卢建远.牦牛NGF基因克隆及其在生殖器官中的表达定位[J].畜牧兽医学报,2022,53(3):778-790. |
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| 作者姓名： | 邬建飞 刘宇 马瑶 蒋旭东 胡双阁 龚三你 字向东 卢建远 |
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| 作者单位： | 西南民族大学 动物科学国家民委重点实验室, 成都 610041 |
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| 基金项目： | 国家重点研发计划项目(2018YFD0502303)；;西南民族大学中央高校基本科研业务专项(2021PTJS26)； |
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| 摘 要： | 旨在探究NGF基因的生物学特性及其在母牦牛生殖器官中的表达特性.本研究收集黄体期母牦牛的心、肝、脾、肺、肾以及胎牛期、卵泡期、黄体期、妊娠期母牦牛的卵巢、子宫和输卵管(n=3),利用RT-PCR克隆牦牛NGF基因,并对其序列进行生物信息学分析,利用RT-qPCR技术分析NGF基因的组织表达特性,利用免疫组化技术(IHC...
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| 关 键 词： | 牦牛 NGF基因 卵巢 组织表达 免疫组化 |
| 收稿时间： | 2021-06-30 |
Cloning of Yak NGF Gene and Its Expression and Localization in Reproductive Organs of Yaks |
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| WU Jianfei,LIU Yu,MA Yao,JIANG Xudong,HU Shuangge,GONG Sanni,ZI Xiangdong,LU Jianyuan.Cloning of Yak NGF Gene and Its Expression and Localization in Reproductive Organs of Yaks[J].Acta Veterinaria et Zootechnica Sinica,2022,53(3):778-790. |
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| Authors: | WU Jianfei LIU Yu MA Yao JIANG Xudong HU Shuangge GONG Sanni ZI Xiangdong LU Jianyuan |
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| Institution: | The Key Laboratory of Animal Science of State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China |
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| Abstract: | The purpose of this study was to explore the molecular characteristics of NGF gene and its expression properties in reproductive organs of female yak (Bos grunniens). The heart, liver, spleen, lung and kidney of female yak at luteal phase were collected. In addition, the ovaries, uterus and oviduct of yaks during fetal, follicular, luteal phases and gestation were collected (n=3). NGF gene was cloned by RT-PCR and its sequence was analyzed by bioinformatics method. The mRNA expression was evaluated by RT-qPCR and NGF protein location in the reproductive organs was detected by immunohistochemistry. The results showed that the CDS region of yak NGF gene was 726 bp, which encoded 241 amino acids. NGF protein belongs to alkaline unstable hydrophilic protein. The results of NGF amino acid phylogenetic tree showed that yaks were first grouped with yellow cattle, and the homology with other species was more than 87%, indicating that NGF gene was relatively conserved in the course of animal evolution. The results of RT-qPCR showed that the expression of NGF gene in ovary was extremely significantly higher than that in heart, liver, kidney, spleen, lung, uterus and oviduct (P<0.01). The relative expression of NGF in ovary of luteal phase was the highest, which was significantly higher than that in fetal, follicular stage and pregnancy (P<0.01). In uterus, the relative expression of NGF was significantly higher during pregnancy than that in fetal and follicular stages (P<0.05). There was no significant difference in the expression of NGF in different physiological stages of oviduct. The immunohistochemistry analysis indicated that NGF protein was mainly localized in granulosa cells and ovarian epithelium cells, with the highest expression in granulosa cells. It was also expressed in yak endometrium and epithelial cells of yak oviduct. In conclusion, the sequence of NGF gene in yak is relatively conserved in the course of evolution, and is highly expressed in the ovary. NGF gene may play an important regulatory role in maintaining ovary function, pregnancy and promoting corpus luteam function in female yaks. |
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| Keywords: | yak nerve growth factor-β(NGF) gene ovary tissue expression immunohistochemistry |
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