牦牛RBM3的基因克隆及其在不同繁殖时期卵巢、输卵管、子宫中的表达定位

doi:10.11843/j.issn.0366-6964.2022.07.017

Abstract

Abstract: The study aimed to investigate the role of RBM3 in the reproductive-related regulation in yak (Bos grunniens). Three healthy female yaks aged 4-6 years old in different reproductive periods (follicular, luteal and pregnancy phases) were used in this study. Nine groups of experimental samples were collected from ovaries, uterus and fallopian tubes, and 3 biological replicates were set in each group. The RBM3 gene of yak was cloned by gene cloning technology and analyzed by bioinformatics. RBM3 relative expression levels of gene and protein in 9 groups of experimental samples were detected by quantitative real-time PCR(qRT-PCR) and Western blot(WB). The distribution of RBM3 protein in samples was detected by immunohistochemistry (IHC). RBM3 gene(GenBank No.:MF142258.1) of yak was cloned in this experiment, it had the closest relationship with wild yak(Bos mutus). Bioinformatics analysis showed that nucleotide No.51 and No.98 were different from several common mammals, and its secondary and tertiary structure was constructed. After prediction, the encoded protein was stable non-transmembrane protein. WB results showed that RBM3 was expressed in fallopian tube, ovary and uterus of yak, but the expression level at luteal phase was significantly higher than that at follicular and pregnancy phases (P<0.05). The expression in fallopian tube at follicular phase was significantly lower than that at luteal and pregnancy phases (P<0.05). The expression in uterus at follicular phase was significantly higher than that at luteal and pregnancy phases(P<0.05). Immunohistochemical results showed that the main expression sites of RBM3 in the ovary were theca follicle, granular layer and luteal cells, while the expression sites in the fallopian tube were epithelium mucosae. For the uterus, the expression sites was mainly in the endometrium and uterine glands. The results of this study suggest that RBM3 may be involved in the regulation of individual estrous cycle and pregnancy process in yak, as well as pregnancy recognition. It provides a reference for the research of RBM3, a cold stress regulatory factor, in the physiological mechanism of yak in cold and hypoxic environment at high altitude, and other related aspects.

Key words: yak, reproductive regulation, RNA-binding motif protein 3, cold stress

CLC Number:

S823.85

ZHANG Hui, PAN Yangyang, WANG Jinglei, ZHANG Rui, HUANG Jiaxin, YU Sijiu, CUI Yan. RBM3 Gene Cloning and Its Expression, Localization in Ovary, Fallopian Tube and Uterus at Different Reproductive Stages of Yak (Bos grunniens)[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2202-2214.

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RBM3 Gene Cloning and Its Expression, Localization in Ovary, Fallopian Tube and Uterus at Different Reproductive Stages of Yak (Bos grunniens)

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