[1]李隐侠,张俊,钱勇,等.湖羊NR5A1基因全序列克隆和表达特征分析[J].江苏农业学报,2019,(01):114-121.[doi:doi:10.3969/j.issn.1000-4440.2019.01.017]
　LI Yin-xia,ZHANG Jun,QIAN Yong,et al.Full sequence cloning and expression characteristics of the ovine NR5A1 gene[J].,2019,(01):114-121.[doi:doi:10.3969/j.issn.1000-4440.2019.01.017]
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湖羊NR5A1基因全序列克隆和表达特征分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年01期
页码:
114-121
栏目:
畜牧兽医·水产养殖
出版日期:
2019-02-26

文章信息/Info

Title:
Full sequence cloning and expression characteristics of the ovine NR5A1 gene
作者:
李隐侠1; 2; 3; 张俊1; 2; 3; 钱勇1; 2; 3; 孟春花1; 2; 3; 王慧利1; 2; 3; 曹少先1; 2; 3; 仲跻峰1; 2; 3
(1.江苏省农业科学院畜牧研究所，江苏南京210014；2.江苏省农业种质资源保护与利用平台，江苏南京210014;3.农业部种养结合重点实验室，江苏南京210014）
Author(s):
LI Yin-xia1; 2; 3; ZHANG Jun1; 2; 3; QIAN Yong1; 2; 3; MENG Chun-hua1; 2; 3; WANG Hui-li1; 2; 3; CAO Shao-xian1; 2; 3; ZHONG Ji-feng1; 2; 3
(1.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.The Jiangsu Provincial Platform for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014, China；3.Key Laboratory of Crop and Animal Integrated Farming, Ministry of Agriculture, Nanjing 210014, China)
关键词:
湖羊; NR5A1基因; 序列特征; 表达
Keywords:
Hu sheep; NR5A1 gene; sequence characterization; expression
分类号:
S826.3
DOI:
doi:10.3969/j.issn.1000-4440.2019.01.017
文献标志码:
A
摘要:
以江苏特色绵羊品种——湖羊为研究对象，克隆湖羊NR5A1基因全序列，分析其序列特征，并对湖羊组织和大、小卵泡中NR5A1表达特征进行研究。结果表明，湖羊NR5A1基因全长19 016 bp，含6个外显子和5个内含子；编码区全长1 386 bp, 编码 461个氨基酸残基，包含经典的DNA结合区（DBD）和配体结合区（LBD）。RT-PCR结果显示NR5A1基因在湖羊组织中广泛表达，其中在卵巢中高表达，脾脏次之；qRT-PCR和Western blot结果显示在湖羊大卵泡中NR5A1 mRNA和蛋白质水平均显著高于小卵泡，表明NR5A1基因参与调控湖羊卵泡发育，并与繁殖性能有关。
Abstract:
Using Hu sheep (Jiangsu featured sheep breed) as the research object, NR5A1 gene full sequences were cloned and the sequence characteristics were analyzed. The expression features of NR5A1 gene in Hu sheep tissues and in large and small follicles were explored. The results showed that the complete sequence of ovine NR5A1 was 19 016 bp in length and contained six exons and five introns. The coding region was 1 386 bp and encoded 461 amino acid residues, including the classical DNA binding domain (DBD) and ligand binding domain (LBD). RT-PCR results showed that NR5A1 gene was widely expressed in Hu sheep tissues, which was highly expressed in ovary, followed by kidney. qRT-PCR and western blot results showed that the expression of NR5A1 in large follicles was significantly higher than that in small follicles, indicating that NR5A1 gene was involved in regulating follicle development and speculating the reproductive performance of Hu sheep.

参考文献/References:

[1]IKEDA Y, LALA DS, LUO X, et al. Characterization of the mouse FTZ-F1 gene, which encodes a key regulator of steroid hydroxylase gene expression [J]. Molecular Endocrinology, 1993, 7(7):852-860.
[2]LIN L, ACHERMANN J C. Steroidogenic factor-1(SF-1, Ad4BP, NR5A1) and disorders of testis development [J]. Sexual Development, 2008, 2(4/5):200-209.
[3]LUO X, IKEDA Y, PARKER K L. A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differentiation [J]. Cell, 1994, 77(4):481-490.
[4]TANTAWY S, LIN L, AKKURT I, et al. Testosterone production during puberty in two 46, XY patients with disorders of sex development and novel NR5A1(SF-1) mutations [J]. European Journal of Endocrinology, 2012, 167(1):125-130.
[5]TAIEB J, GRYNBERG M, PIERRE A, et al. FSH and its second messenger cAMP stimulate the transcription of human anti-Müllerian hormone in cultured granulosa cells [J]. Molecular Endocrinology, 2011, 25(4):645-655.
[6]MATΜLIS C K, MAYO K E. The LIM domain protein FHL2 interacts with the NR5A family of nuclear receptors and CREB to activate the inhibin-α subunit gene in ovarian granulosa cells [J]. Molecular Endocrinology, 2012, 26(8): 1278-1290.
[7]MURAYAMA C, MIYAZAKI H, MIYAMOTO A, et al. Luteinizing hormone (LH) regulates production of androstenedione and progesterone via control of histone acetylation of StAR and CYP17 promoters in ovarian theca cells [J]. Molecular Cellular Endocrinology, 2012, 350(1): 1-9.
[8]IMAMICHI Y, MIZUTANI T, JU Y, et al. Transcriptional regulation of human ferredoxin 1 in ovarian granulosa cells [J]. Molecular Cellular Endocrinology, 2013, 370(1/2): 1-10.
[9]LAW NC, WECK J, KYRISS B, et al. Lhcgr expression in granulosa cells: roles for PKA-phosphorylated β-catenin, TCF3, and FOXO1 [J]. Molecular Endocrinology, 2013, 27(8):1295-1310.
[10]YIN M, L M, YAO G, et al. Transactivation of microRNA-383 by steroidogenic factor-1 promotes estradiol release from mouse ovarian granulosa cells by targeting RBMS1 [J]. Molecular Endocrinology, 2012, 26(7): 1129-1143.
[11]FORTIN J, KUMAR V, ZHOU X, et al. NR5A2 regulates Lhb and Fshb transcription in gonadotrope-like cells in vitro, but is dispensable for gonadotropin synthesis and fertility in vivo [J]. PLoS ONE, 2013, 8(3):e59058
[12]SCHANG A L, QURAT B, SIMON V, et al. Mechanisms underlying the tissue-specific and regulated activity of the Gnrhr promoter in mammals [J]. Front Endocrinol (Lausanne), 2012, 3: 162.
[13]李隐侠，张俊，钱勇，等. 湖羊NR5A1基因SNPs筛选及其与产羔数的关联分析 [J]. 江苏农业学报, 2017, 33（1）：124-132.
[14]TAMURA K, PETERSON D, PETERSON N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods [J]. Molecular Biology Evolution, 2011, 28(10):2731-2739.
[15]YAO Y, REHEMAN A, XU Y, et al. miR-125b contributes to ovarian granulosa cell apoptosis through targeting BMPR1B, a major gene for sheep prolificacy [J]. Reproduction Science, 2018. doi: 10.1177/1933719118770544 .
[16]OBA K, YANASE T, NOMURA M, et al. Structural characterization of human Ad4bp (SF-1) gene [J]. Biochemical Biophysical Research Communication, 1996, 226(1):261-267.
[17]YUMOTO F, NGUYEN P, SABLIN E P, et al. Structural basis of coactivation of liver receptor homolog-1 by β-catenin [J]. Proceedings of the National Academy of Sciences of USA, 2012, 109(1): 143-148.
[18]LI Y X, ZHANG J, QIAN Y, et al. Molecular characterization, expression, polymorphism of NR5A2 and its relationship with litter size in Hu sheep [J]. Genetics and Molecular Research,2015, 14(4):12765-12775.
[19]MLYNARCZUK J, WROBEL M H, ZIOLKOWSKA A, et al. Involvement of the orphan nuclear receptor SF-1 in the effect of PCBs, DDT and DDE on the secretion of steroid hormones and oxytocin from bovine granulosa cells [J]. Animal Reproduction Science, 2013, 143(1-4): 30-37.
[20]BUAAS F W, GARDINER J R, CLAYTON S, et al. In vivo evidence for the crucial role of SF1 in steroid-producing cells of the testis, ovary and adrenal gland [J]. Development, 2012, 139(24): 4561-4570.
[21]EL-KHAIRI R, MARTINEZ-AGUAYO A, FERRAZ-DE-SOUZA B, et al. Role of DAX-1 (NR0B1) and steroidogenic factor-1 (NR5A1) in human adrenal function [J]. Endocrine Development, 2011, 20:38-46.
[22]EHRLUND A, JONSSON P, VEDIN L L, et al. Knockdown of SF-1 and RNF31 affects components of steroidogenesis, TGFβ, and Wnt/β-catenin signaling in adrenocortical carcinoma cells [J]. PLoS ONE, 2013, 7(3):e32080.
[23]FALENDER A E, LANZ R, MALENFANT D, et al. Differential expression of steroidogenic factor-1 and FTF/LRH-1 in the rodent ovary [J]. Endocrinology, 2013, 144(8):3598-3610.
[24]JIN X, HAN C S, ZHANG X S, et al. Stem cell factor modulates the expression of steroidogenesis related proteins and FSHR during ovarian follicular development [J]. Fronters in Bioscience: a Journal and Virtual Library, 2005, 10:1573-1580.
[25]YIN M, WANG X, YAO G, et al. Transactivation of micrornA-320 by microRNA-383 regulates granulosa cell functions by targeting E2F1 and SF-1 proteins [J]. The Journal of Biological Chemistry, 2014, 289(26):18239-18257.
[26]WANG L, LI C, LI R, et al. MicroRNA-764-3p regulates 17β-estradiol synthesis of mouse ovarian granulosa cells by targeting steroidogenic factor-1 [J]. In Vitro Cellular & Developmental Biology Animal, 2016, 52(3):365-373.
[27]PELUSI C, IKEDA Y, ZUBAIR M, et al. Impaired follicle development and infertility in female mice lacking steroidogenic factor 1 in ovarian granulosa cells [J]. Biology of Reproduction, 2008, 79(6): 1074 -1083.

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备注/Memo

备注/Memo:
收稿日期：2018-04-27 基金项目：江苏省自然科学基金项目（BK20140750）；国家自然科学基金项目（31501934）；苏州市科技支撑计划项目(SNG201416) 作者简介：李隐侠(1979-)，女，河南固始人，博士，副研究员，主要从事动物遗传育种与繁殖方面的研究。（Tel）025-84390095; (Email) liyxmh@126.com 通讯作者：曹少先， (E-mail) caoshaoxian@163.com
更新日期/Last Update: 2019-02-27