Cloning of Tibetan Sheep (Ovis aries) TPI1 Gene and Its Expression in Male Reproductive Organs
ZHANG Hong-Yu1, LI Tao-Tao1, MA You-Ji1,2,*, WANG Xia1, YIN De-En1, ZHAO Xing-Xu3, LI Dong-Hong4
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 Sheep Breeding Biotechnology Engineering Laboratory of Gansu Province, Minqin 733300, China; 3 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 4 Animal Husbandry and Veterinary Station of Renliu of Lintong District in Xi'an City, Xi'an 710600, China
Abstract Sertoli cells can provide energy substrate for the development of male germ cells through glycolysis. Triosephosphate isomerase 1 (TPI1) is a key enzyme in glycolysis. In order to explore the biological function of TPI1 gene in the development of reproductive organs of male Tibetan sheep (Ovis aries), the cDNA sequence of TPI1 gene (GenBank No. MN847717) was cloned, and its expression and localization patterns in developmental testis and epididymis were detected by real time quantitative PCR (qRT-PCR), Western blot and immunohistochemistry. The results showed that the length of CDS region of Tibetan sheep TPI1 gene was 861 bp and encoded 286 amino acids, showing high degrees of homology with other mammals. TPI1 gene and its protein were expressed in testis and epididymis in all developmental stages, but mainly in testes and epididymides from 1-year-old and 3-year-old sheep. TPI1 protein was mainly distributed in Sertoli cells within testes at all stages of development, and sperm tails within epididymides from post-puberty (1-year-old and 3-year-old). In addition, weak TPI1 protein signals were present in Leydig cells, spermatogonia and spermatocytes from testes, and pseudostratified columnar ciliated epitheliums and loose connective tissues from epididymides. These results taken together demonstrated that TPI1 gene might be involved in the regulation of glycolysis in Sertoli cells and sperm flagella, as well as the secretory function of pseudostratified columnar ciliated epithelium of epididymis, which in turn might promote the formation and maturation of sperms. This study provides basic materials for further investigation on the regulatory mechanism of TPI1 gene in ovine and other mammalian spermatogenesis.
ZHANG Hong-Yu,LI Tao-Tao,MA You-Ji, et al. Cloning of Tibetan Sheep (Ovis aries) TPI1 Gene and Its Expression in Male Reproductive Organs[J]. 农业生物技术学报, 2020, 28(6): 1012-1020.
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