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.
张宏豫, 李讨讨, 马友记, 王霞, 尹德恩, 赵兴绪, 李东红. 藏绵羊TPI1基因的克隆及其在雄性生殖器官中的表达[J]. 农业生物技术学报, 2020, 28(6): 1012-1020.
ZHANG Hong-Yu, LI Tao-Tao, MA You-Ji, WANG Xia, YIN De-En, ZHAO Xing-Xu, LI Dong-Hong. Cloning of Tibetan Sheep (Ovis aries) TPI1 Gene and Its Expression in Male Reproductive Organs. 农业生物技术学报, 2020, 28(6): 1012-1020.
[1] 窦露, 王炜, 董小龙, 等. 2018. 山羊钴胺素转运蛋白受体基因CD320的克隆与表达模式[J]. 农业生物技术学报, 26(08): 1342-1350. (Dou L, Wang W, Dong X L, et al.2018. Cloning and expression patterns of cobalamin transporter receptor gene CD320 in goat (Capra hircus)[J]. Journal of Agricultural Biotechnology, 26(08): 1342-1350.) [2] 李玉华, 付杰丽, 李培飞, 等. 2017. 金黄色葡萄球菌对体外保存猪精子活力及蛋白磷酸化修饰的影响[J]. 畜牧兽医学报, 48(09): 1665-1673. (Li Y H, Fu J L, Li P F, et al.2017. Effects of on motility and protein phosphorylation modification of boar sperm during storage in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 48(09): 1665-1673.) [3] 宋慧子. 2019. 永生化绵羊附睾上皮细胞系的建立及其生物学特性分析[D]. 硕士学位论文, 内蒙古农业大学, 导师: 张家新, pp. 1-4. (Song H Z.2019. Establishment of sheep immortalized epididymal epithelial cell line and analysis of its biological characteristics[D]. Thesis for M.S., Inner Mongolia Agricultural University, Supervisor: Zhang J X, pp. 1-4.) [4] Auer J, Camoin L, Courtot A, et al.2004. Evidence that P36, a human sperm acrosomal antigen involved in the fertilization process is triosephosphate isomerase[J]. Molecular Reproduction Development, 68(4): 515-523. [5] Bone W, Cooper T.2000. In vitro inhibition of rat cauda epididymal sperm glycolytic enzymes by ornidazole, alpha-chlorohydrin and 1-chloro-3-hydroxypropanone[J]. International Journal of Andrology, 23(5): 284-293. [6] Byrne K, Leahy T, McCulloch R, et al.2012. Comprehensive mapping of the bull sperm surface proteome[J]. Proteomics, 12(23-24): 3559-3579. [7] Hermo L, Oliveira R, Smith C, et al.2019. Dark side of the epididymis: Tails of sperm maturation[J]. Andrology, 7(5): 566-580. [8] Hu Y, Deng J, Tian K, et al.2019. MiR-8-3p regulates hyperthermia-induced lactate secretion by targeting PPP2R5B in boar Sertoli cells[J]. Molecular Reproduction Development, 86(11): 1720-1730. [9] Ijiri T, Vadnais M, Lin A, et al.2013. Male mice express spermatogenic cell-specific triosephosphate isomerase isozymes[J]. Molecular Reproduction Development, 80(10): 862-870. [10] Jiang H, Ma N, Shang Y, et al.2017. Triosephosphate isomerase 1 suppresses growth, migration and invasion of hepatocellular carcinoma cells[J]. Biochemical Biophysical Research Communications, 482(4): 1048-1053. [11] Lehti M, Sironen A.2017. Formation and function of sperm tail structures in association with sperm motility defects[J]. Biology of Reproduction, 97(4): 522-536. [12] Levine A, Puzio-Kuter A.2010. The control of the metabolic switch in cancers by oncogenes and tumor suppressor genes[J]. Science, 330(6009): 1340-1344. [13] Li J, Zhang J, Xie F, et al.2018. Macrophage migration inhibitory factor promotes Warburg effect via activation of the NF-κB/HIF-1α pathway in lung cancer[J]. International Journal of Molecular Medicine, 41(2): 1062-1068. [14] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J]. Methods, 25(4): 402-408. [15] Lu Z, Ma Y, Zhang Q, et al.2018. Proteomic analyses of ram (Ovis aries) testis during different developmental stages[J]. Animal Reproduction Science, 189: 93-102. [16] Martins A, Alves M, Simões V, et al.2013. Control of Sertoli cell metabolism by sex steroid hormones is mediated through modulation in glycolysis-related transporters and enzymes[J]. Cell and Tissue Research, 354(3): 861-868. [17] Ni F, Hao S, Yang W.2019. Multiple signaling pathways in Sertoli cells: Recent findings in spermatogenesis[J]. Cell Death Disease, 10(8): 541. [18] Oliveira P F, Martins A D, Moreira A C, et al.2015. The Warburg effect revisited-lesson from the Sertoli cell[J]. Medicinal Research Reviews, 35(1): 126-151. [19] Rato L, Socorro S, Cavaco J E B, et al.2010. Tubular fluid secretion in the seminiferous epithelium: Ion transporters and aquaporins in Sertoli cells[J]. The Journal of Membrane Biology, 236(2): 215-224. [20] Takei G, Miyashiro D, Mukai C, et al.2014. Glycolysis plays an important role in energy transfer from the base to the distal end of the flagellum in mouse sperm[J]. The Journal of Experimental Biology, 217: 1876-1886. [21] Yang L, Shen J, Chen J, et al.2019. Reduced glycolysis contributed to inhibition of testis spermatogenesis in rats after chronic methamphetamine exposure[J]. Medical Science Monitor, 25: 5453-5464. [22] Yu J, Sun J, Fan Y, et al.2019. Exposure to Pb and Cd alters MCT4/CD147 expression and MCT4/CD147-dependent lactate transport in mice Sertoli cells cultured in vitro[J]. Toxicology in Vitro, 56: 30-40.
