Expression and Localization of LHR and PRLR Genes in Testicles of Yak (Bos grunniens) at Different Developmental Stages
MA Fu-Long1,2, CHU Min1, CHANG Yong-Fang1,2, BAO Peng-Jia1, GUO Xian1, Qudratullah Kalwar1, MA Qi-Cai1,2, PAN He-Ping2, YAN Ping1*
1 Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Aicultural Sciences/ Gansu Key Laboratory for Yak Breeding Engineering of Gansu Province, Lanzhou 730050, China; 2 College of Life Science and Engineering, Northwest University for Nationlities, Lanzhou 730030, China
Abstract：Luteinizing hormone receptor (LHR) and prolactin receptor (PRLR) genes play essential roles in mammalian reproductive development. But still no any available research regarding these genes in yak (Bos grunniens). Therefore this study was planned to explore the role of LHR and PRLR genes in different tissues of Yak during different developmental ages (6, 24, 36 and 72 months). The mRNA expression levels of LHR and PRLR in the testis tissues of yak at different developmental stages and organs of yak were detected by qRT-PCR, while the protein expression of LHR and PRLR were evaluated by immunohistochemistry.The results showed that mRNA expression levels of LHR and PRLR genes in yak testis increased significantly from infancy to sexual maturity (P<0.01), but there was no significant difference between adult and sexual maturity. The expression level of LHR gene mRNA in different organs of adult yak was higher in testis than in kidney, skeletal muscle and lung (P<0.05). While the expression level of PRLR mRNA was higher in kidney, followed by skeletal muscle, testis and spleen (P<0.05). Additionally, immunohistochemistry findings revealed that LHR and PRLR proteins were located in testicular supporting cells, mesenchymal cells, primary spermatocytes and secondary spermatocytes, and they were increased gradually from infancy to sexual maturity stage and tended to be stable in adulthood. These findings displayed that LHR and PRLR may be vital for sperm maturation, in the biological functions of interstitial cells, and in regulating the development of other non-gonadal tissues. These results provide an important theoretical basis for revealing the molecular mechanism of testis development and reproductive regulation in yaks.
马富龙, 褚敏, 常永芳, 包鹏甲, 郭宪, Qudratullah Kalwar, 马启财, 潘和平, 阎萍. LHR、PRLR基因在不同发育阶段牦牛睾丸中的表达与定位[J]. 农业生物技术学报, 2020, 28(4): 672-680.
MA Fu-Long, CHU Min, CHANG Yong-Fang, BAO Peng-Jia, GUO Xian, Qudratullah Kalwar, MA Qi-Cai, PAN He-Ping, YAN Ping. Expression and Localization of LHR and PRLR Genes in Testicles of Yak (Bos grunniens) at Different Developmental Stages. 农业生物技术学报, 2020, 28(4): 672-680.
[1] 陈晓燕, 温海深, 何峰, 等. 2010. 半滑舌鳎促黄体激素受体基因片段的克隆及组织表达分析[J]. 中国海洋大学学报(自然科学版), 40(3): 71-77. (Chen X Y, Wen H S, He F, et al.2010. Partial sequence cloning of LHR gene in Cynoglossus semilaevis and its tissue expression analysis[J]. Journal of Ocean University of China (Natural Science Edition), 40(3): 71-77.) [2] 韩杰, 熊显荣, 王艳, 等. 2018. 牦牛KDM1A基因克隆及其在不同发育时期睾丸中的表达规律[J]. 畜牧兽医学报, 49(12): 248-256. (Han J, Xiong X R, Wang Y, et al.2018. Cloning of yak KDM1A gene and its expression profile in testis during different developmental stages[J]. Acta Veterinaria et Zootechnica Sinica, 49(12): 248-256.) [3] 李晓雨, 贺小云, 刘秋月, 等. 2018. TAC1和PRLR基因在绵羊不同繁殖状态下的表达模式分析[J]. 畜牧兽医学报, 49(2): 253-262. (Li X Y, He X Y, Liu Q Y, et al.2018. Expression pattern analysis of TAC1 and PRLR genes in different reproductive states of sheep[J]. Acta Veterinaria et Zootechnica Sinica, 49(2): 253-262.) [4] 牛淑玲, 丰培金, 周虚. 2003. 促黄体素受体研究进展[J]. 动物医学进展, 24(5). (Niu S L, Feng P J, Zhou X, et al.2003. Advances on the luteinizing hormone receptor[J]. Progress in Veterinary Medicine, 24(5).) [5] 邵康, 周杰, 吴小雪, 等. 2011. 猪睾丸中脂联素受体与LHR、CYP11A1、StAR基因表达的发育变化及其相关性研究[J]. 畜牧兽医学报, 42(12): 1680-1685. (Shao K, Zhou J, Wu X X, et al.2011. The developmental patterns and correlation of adiponectin receptors, LHR, CYP11A1 and StAR mRNA expression in testis of wan nan hua pigs[J]. Acta Veterinaria et Zootechnica Sinica, 42(12): 1680-1685.) [6] 谢海燕. 2011. 黑线仓鼠LHR部分序列克隆及组织器官的表达差异[D]. 硕士学位论文, 曲阜师范大学, 导师: 徐来祥, pp. 1-3. (Xie H Y.2011. Partial sequence cloning and tissue and organ expression difference of black line hamster LHR[D]. Thesis for M.S., Qufu Normal University, Supervisor: Xu L X, pp. 1-3.) [7] 邢光东. 2011. 鹅催乳素受体基因克隆和多克隆抗体制备及组织表达分析[D]. 博士学位论文, 南京农业大学, 导师: 王根林, pp. 79-80. (Xing G D.2011. Gene cloning and polycolonal antibody preparation and tissue expression analysis of prolactin receptor in the goose[D]. Thesis for Ph.D., Nanjing Agricultural University, Supervisor: Wang G L. pp, 79-80.) [8] 曾献存, 陈韩英, 贾斌, 等. 2011. 中国美利奴羊PROP1和PRLR基因的表达分析[J]. 畜牧兽医学报, 42(08): 1057-1062. (Zeng X C, Chen H Y, Jia B, et al.2011. The expression analysis of PROPl and PRLR genes in Chinese merino sheep[J]. Acta Veterinaria et Zootechnica Sinica, 42(08): 1057-1062.) [9] Abdallah M A, Lei Z M, Li X, et al.2004. Human fetal nongonadal tissues contain human chorionic gonadotropin/luteinizing hormone receptors[J]. The Journal of Clinical Endocrinology & Metabolism, 89(2). [10] Bazan J F.1990. Structural design and molecular evolution of a cytokine receptor superfamily[J]. Proceedings of the National Academy of Sciences, 87(18): 6934-6938. [11] Cassy S, Charlier M, L Bélair, et al.2000. Increase in prolactin receptor (PRL-R) mRNA level in the mammary gland after hormonal induction of lactation in virgin ewes[J]. Domestic Animal Endocrinology, 18(1): 41-55. [12] Cosenza G, Iannaccone M, Auzino B, et al.2018. Remarkable genetic diversity detected at river buffalo prolactin receptor (PRLR) gene and association studies with milk fatty acid composition[J]. Animal Genetics, 49(3). [13] Exbrayat J M, Gérard Morel.2003. Visualization of gene expression of prolactin-receptor (PRL-R) by in situ hybridization in reproductive organs of Typhlonectes compressicauda, a gymnophionan amphibian[J]. Cell & Tissue Research, 312(3): 361-367. [14] Fields M J, Shemesh M, Fields M J.2004. Extragonadal luteinizing hormone receptors in the reproductive tract of domestic animals[J]. Biology of Reproduction, 71(5): 1412. [15] Freeman M E, Kanyicska Béla, Anna L, et al.2000. Prolactin: Structure, function, and regulation of secretion[J]. Physiological Reviews, 80(4): 1523-1631. [16] Hair W M.2002. Prolactin receptor expression in human testis and accessory tissues: Localization and function[J]. Molecular Human Reproduction, 8(7): 606-611. [17] Kelly P A, Djiane J, Postelvinay M C, et al.1991. The prolactin/growth hormone receptor family[J]. Endocrine Reviews, 12(3): 235-51. [18] Lei Z M.1993. Novel expression of human chorionic gonadotropin/luteinizing hormone receptor gene in brain[J]. Endocrinology, 132(5): 2262-2270. [19] Lei Z M, Mishra S, Zou W, et al.2001. Targeted disruption of luteinizing hormone/human chorionic gonadotropin receptor gene[J]. Molecular Endocrinology, 15(1): 184-200. [20] Noh, Eon G, Kim, et al.2013. Characterization of genes encoding prolactin and prolactin receptors in starry flounder Platichthys stellatus and their expression upon acclimation to freshwater[J]. Fish Physiology & Biochemistry, 39(2): 263-275. [21] Ozaki Y, Damsteegt E L, Setiawan A N, et al.2019. Expressional regulation of gonadotropin receptor genes and androgen receptor genes in the eel testis[J]. General and Comparative Endocrinology, 280(9): 123-133. [22] Pabon J E, Li X, Lei Z M, et al.Novel presence of luteinizing hormone/chorionic gonadotropin receptors in human adrenal glands[J]. The Journal of Clinical Endocrinology & Metabolism, 81(6): 2397-2400. [23] Picazo R A, Garcia Ruiz J P, Santiago Moreno J, et al.2004. Cellular localization and changes in expression of prolactin receptor isoforms in sheep ovary throughout the estrous cycle[J]. Reproduction, 128(5): 545-553. [24] Pratt S L, Calcatera S M, Stowe H M, et al.2015. Identification of bovine prolactin in seminal fluid, and expression and localization of the prolactin receptor and prolactin-inducible protein in the testis and epididymis of bulls exposed to ergot alkaloids[J]. Theriogenology, 83(4): 662-669. [25] Qiu Q, Zhang G, Ma T, et al.2012. The yak genome and adaptation to life at high altitude[J]. Nature Genetics, 44(8): 946-9. [26] Rao C V.2001. Multiple novel roles of luteinizing hormone[J]. Fertility & Sterility, 76(6): 1097-1100. [27] Rhee J S, Kim R O, Seo J S, et al.2010. Effects of salinity and endocrine-disrupting chemicals on expression of prolactin and prolactin receptor genes in the euryhaline hermaphroditic fish, Kryptolebias marmoratus[J]. Comparative Biochemistry and Physiology, Part C, 152(4): 413-423. [28] Stocco D.1998. Recent advances in the role of StAR[J]. Reviews of Reproduction, 3(2): 82-85. [29] Stocco D M.2000. StARTing to understand cholesterol transfer[J]. Nature Structural Biology, 7(6): 445. [30] Tomy S, Chang Y M, Chen Y H, et al.2009. Salinity effects on the expression of osmoregulatory genes in the euryhaline black porgy Acanthopagrus schlegeli[J]. General and Comparative Endocrinology, 161(1): 123-132. [31] Yuan L J, Peng C, Liu B H, et al.2019. Identification and characterization of a luteinizing hormone receptor (LHR) Homolog from the chinese mitten crab eriocheir sinensis[J]. International Journal of Molecular Sciences, 20(7).
