Cloning of GNAQ Gene in Yak (Bos grunniens) and Its Expression in Female Reproductive Axis
CHEN Yi-Wei1,2,3, CHU Min1,3, ZHANG Ren-Zheng1,3, XIE Jian-Peng1,3, MA Lan-Hua1,2,3, PAN He-Ping2, YAN Ping1,3,*
1 Lanzhou Institute of Husbandry and Pharmaceutical Sciences/Key Laboratory of Yak Breeding Engineering Gansu Province, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China; 2 College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China; 3 Key Laboratory of Animal Genetics and Breeding on Tibetan Plateau, Ministry of Agriculture and Rural Affairs, Lanzhou 730050, China
Abstract：G protein subunit alpha Q (GNAQ) regulate G protein-coupled receptors and mediate downstream signaling pathways through encoding Gαq protein, which involves many physiological processes such as nutrient metabolism, reproduction regulation and cell signal transduction. This study aimed to explore the expression of GNAQ in reproductive axons of female yak (Bos grunniens). The samples of hypothalamus, pituitary and ovarian of healthy female yaks (adult) during follicular phase were collected. The CDS of GNAQ gene was cloned with yak ovarian cDNA as template by reverse transcription-PCR (RT-PCR) and it was analyzed by bioinformatics software. The hypothalamic-pituitary-ovarian reproductive axis (HPOA) expression characteristics of yak GNAQ gene and protein levels were detected by qRT-PCR, Western blot and immunohistochemistry (IHC) .The results showed that the CDS of yak GNAQ gene (GenBank No. OP811271) was 1 080 bp in length and encoded 359 amino acids. GNAQ was close to amino acid homology of ordinary cattle (B. taurus), and distant to mouse (Mus musculus), and it was highly conserved in the evolution of the species. It was found that GNAQ gene and protein levels were expressed in hypothalamus, pituitary and ovary of yak by qRT-PCR and Western blot, and the highest expression level was found in pituitary, the expression level in hypothalamus was significantly higher than in ovary (P<0.05). Immunohistochemistry results showed that GNAQ protein was strongly positive in ovarian granulosa cells and was mainly expressed in cytoplasm, which was consistent with the prediction of subcellular localization. This study provides theoretical basis for further exploration of the role of GNAQ gene in reproductive physiology regulation of yak, which is helpful to study the regulation and mechanism of seasonal estrus in yak.
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