绵羊FSHR基因生物信息学分析及其器官表达规律研究
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摘要
为了验证及进一步研究促卵泡素受体(follicle-stimulating hormone receptor,FSHR)基因在绵羊繁殖力中的作用机制,试验采用生物信息学方法分析了绵羊FSHR基因结构、FSHR蛋白性质和结构、同源性、遗传进化、信号通路、蛋白互作网络和器官表达规律。结果表明:绵羊FSHR DNA全长196 149 bp,而编码蛋白质的mRNA序列全长只有2 431 bp,FSHR基因含有10个外显子;FSHR基因编码695个氨基酸,FSHR蛋白属于不稳定的脂溶性蛋白;FSHR蛋白是有7个跨膜结构的膜受体,包含LRR_5、GnRH_trans、7tm_1三种功能结构域,这三种结构域分别行使不同功能,共同激活G蛋白偶联机制,激活腺苷酸环化酶,使细胞内CAMP或Ca~(2+)内流增加,从而对细胞的生命活动进行调节;FSHR基因不但与繁殖相关基因Gdf9、Bmp15有关,还与抑制肿瘤转移基因Kiss1有关;FSHR的同源性分析和进化树构建结果显示,绵羊和山羊亲缘关系最近,其次是牛,与斑马鱼亲缘关系最远;FSHR基因在绵羊睾丸和卵巢中特异性高表达,并在各物种间的表达具有一致性。说明FSHR基因表达可能受内含子极为复杂和重要的调控,且在繁殖中起重要作用。该基因在睾丸中特异性高表达,与促卵泡生成素(FSH)对雄性动物曲细精管的发育、成熟和精子生成作用相吻合。
In order to verify and further study the mechanism of FSHR gene in sheep fertility, the bioinformatics method was used to analyze the genetic structure, protein structure and physicochemical properties, homology, genetic evolution, signal pathway, protein interaction network and tissue expression pattern of sheep FSHR. The results showed that the length of sheep FSHR DNA was 196 149 bp, while the full length of mRNA encoding protein was only 2 431 bp. The FSHR gene contained 10 exons, the FSHR gene encoded 695 amino acids, and the FSHR protein was an unstable liposoluble protein. FSHR protein was a membrane receptor with seven transmembrane structures. It contained three types of functional domains LRR_5, GnRH_trans, and 7 tm_1. These three types of domains respectively performed different functions and activate the G protein coupling mechanism to activate adenylyl cyclase to increase intracellular CAMP or increase Ca~(2+) influx, which regulated the life activities of the cell. The FSHR gene was not only associated with the reproduction-related genes Gdf9 and Bmp15, but also related to the tumor metastasis suppressor gene Kiss1. Homology analysis and phylogenetic tree construction of FSHR showed that sheep and goats had the closest relationship, followed by cattle. And it was farthest from zebrafish. The FSHR gene was highly expressed specifically in the testes and ovaries of sheep and was conservative among species. The result suggests that FSHR gene expression may be regulated by introns, the process is very complex and important. The high expression of this gene in testis coincides with FSH promoting the development, maturation and spermatogenesis of seminiferous tubules in male animals.
In order to verify and further study the mechanism of FSHR gene in sheep fertility, the bioinformatics method was used to analyze the genetic structure, protein structure and physicochemical properties, homology, genetic evolution, signal pathway, protein interaction network and tissue expression pattern of sheep FSHR. The results showed that the length of sheep FSHR DNA was 196 149 bp, while the full length of mRNA encoding protein was only 2 431 bp. The FSHR gene contained 10 exons, the FSHR gene encoded 695 amino acids, and the FSHR protein was an unstable liposoluble protein. FSHR protein was a membrane receptor with seven transmembrane structures. It contained three types of functional domains LRR_5, GnRH_trans, and 7 tm_1. These three types of domains respectively performed different functions and activate the G protein coupling mechanism to activate adenylyl cyclase to increase intracellular CAMP or increase Ca~(2+) influx, which regulated the life activities of the cell. The FSHR gene was not only associated with the reproduction-related genes Gdf9 and Bmp15, but also related to the tumor metastasis suppressor gene Kiss1. Homology analysis and phylogenetic tree construction of FSHR showed that sheep and goats had the closest relationship, followed by cattle. And it was farthest from zebrafish. The FSHR gene was highly expressed specifically in the testes and ovaries of sheep and was conservative among species. The result suggests that FSHR gene expression may be regulated by introns, the process is very complex and important. The high expression of this gene in testis coincides with FSH promoting the development, maturation and spermatogenesis of seminiferous tubules in male animals.
引文
[1] 朱桂金,聂睿.FSH和LH的作用[J]. 生殖医学杂志, 2010, 19(4):353-355.
[2] 华荣茂,曾斌. FSH研究进展[J]. 江西科技师范大学学报, 2014(6):7-12.
[3] MARSTERS P, KENDALL N R, CAMPBELL B K. Temporal relationships between FSH receptor, type 1 insulin-like growth factor receptor, and aromatase expression during FSH-induced differentiation of bovine granulosa cells maintained in serum-free culture[J]. Mol Cell Endocrinol, 2003, 203(1): 117-127.
[4] 韦雪芳, 王冬梅, 刘思, 等. 信号肽及其在蛋白质表达中的应用[J]. 生物技术通报, 2006(6): 38-42.
[5] DAELEMANS C, SMITS G, DE MAERTELAER V, et al. Prediction of severity of symptoms in iatrogenic ovarian hyperstimulation syndrome by follicle-stimulating hormone receptor Ser680Asn polymorphism[J]. J Clin Endocrinol Metab, 2004, 89(12): 6310-6315.
[6] GEORGE L, ARNAU C, LEONARDO P. The G-protein coupled receptor family: actors with many faces[J]. Curr Pharm Des, 2012, 18(2): 175-185.
[7] 刘映, 李军, 张跃环, 等. 香港牡蛎新型LRR受体克隆与功能分析[J]. 水产学报, 2017, 41(3): 347-357.
[8] KRISHNAN A, ALMEN M S, FREDRIKSSON R, et al. The origin of GPCRs: identification of mammalian like rhodopsin, adhesion, glutamate and frizzled GPCRs in fungi[J]. PLoS One, 2012, 7(1): e29817.
[9] 石燕, 郁琳, 朱燕, 等. 以FSH受体的配体结合部位作为抗原进行主动免疫对大鼠生殖的影响[J]. 实验动物与比较医学, 2013, 33(1):12-18.
[10] 李莉莉. 两种新的FSHR基因mRNA选择性剪切体的鉴定及功能的初步研究[D]. 杭州:浙江大学, 2013.
[11] 覃玉凤, 李维, 陈瑶生, 等. BMP15 基因研究进展[J]. 农业生物技术学报, 2017, 25(2): 324-334.
[12] 顾祝荣, 郭海燕, 王强, 等. 湖羊 GDF9 基因多态性和产羔性能的关系羔[J]. 安徽农业科学, 2018, 46(6): 87-89.
[13] 陈祥, 龙威海, 孙振梅, 等. 贵州地方山羊 FSHR 基因与繁殖性状的相关性研究[J]. 农业生物技术学报, 2017, 25(1): 94-101.
[14] 陈志华, 林素勇, 韩宏景, 等. 慢病毒转染 KISS1 基因对人结直肠癌 HCT116 细胞增殖, 侵袭和迁移能力的影响[J]. 吉林大学学报(医学版), 2017, 43(3): 577-581.
[2] 华荣茂,曾斌. FSH研究进展[J]. 江西科技师范大学学报, 2014(6):7-12.
[3] MARSTERS P, KENDALL N R, CAMPBELL B K. Temporal relationships between FSH receptor, type 1 insulin-like growth factor receptor, and aromatase expression during FSH-induced differentiation of bovine granulosa cells maintained in serum-free culture[J]. Mol Cell Endocrinol, 2003, 203(1): 117-127.
[4] 韦雪芳, 王冬梅, 刘思, 等. 信号肽及其在蛋白质表达中的应用[J]. 生物技术通报, 2006(6): 38-42.
[5] DAELEMANS C, SMITS G, DE MAERTELAER V, et al. Prediction of severity of symptoms in iatrogenic ovarian hyperstimulation syndrome by follicle-stimulating hormone receptor Ser680Asn polymorphism[J]. J Clin Endocrinol Metab, 2004, 89(12): 6310-6315.
[6] GEORGE L, ARNAU C, LEONARDO P. The G-protein coupled receptor family: actors with many faces[J]. Curr Pharm Des, 2012, 18(2): 175-185.
[7] 刘映, 李军, 张跃环, 等. 香港牡蛎新型LRR受体克隆与功能分析[J]. 水产学报, 2017, 41(3): 347-357.
[8] KRISHNAN A, ALMEN M S, FREDRIKSSON R, et al. The origin of GPCRs: identification of mammalian like rhodopsin, adhesion, glutamate and frizzled GPCRs in fungi[J]. PLoS One, 2012, 7(1): e29817.
[9] 石燕, 郁琳, 朱燕, 等. 以FSH受体的配体结合部位作为抗原进行主动免疫对大鼠生殖的影响[J]. 实验动物与比较医学, 2013, 33(1):12-18.
[10] 李莉莉. 两种新的FSHR基因mRNA选择性剪切体的鉴定及功能的初步研究[D]. 杭州:浙江大学, 2013.
[11] 覃玉凤, 李维, 陈瑶生, 等. BMP15 基因研究进展[J]. 农业生物技术学报, 2017, 25(2): 324-334.
[12] 顾祝荣, 郭海燕, 王强, 等. 湖羊 GDF9 基因多态性和产羔性能的关系羔[J]. 安徽农业科学, 2018, 46(6): 87-89.
[13] 陈祥, 龙威海, 孙振梅, 等. 贵州地方山羊 FSHR 基因与繁殖性状的相关性研究[J]. 农业生物技术学报, 2017, 25(1): 94-101.
[14] 陈志华, 林素勇, 韩宏景, 等. 慢病毒转染 KISS1 基因对人结直肠癌 HCT116 细胞增殖, 侵袭和迁移能力的影响[J]. 吉林大学学报(医学版), 2017, 43(3): 577-581.