牦牛NGF基因克隆及其在生殖器官中的表达定位

doi:10.11843/j.issn.0366-6964.2022.03.011

Abstract

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.

Key words: yak, nerve growth factor-β(NGF) gene, ovary, tissue expression, immunohistochemistry

CLC Number:

S823.85

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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Cloning of Yak NGF Gene and Its Expression and Localization in Reproductive Organs of Yaks

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