Tissue Expression, Functional Prediction of CYP17A1 and CYP19A1 Genes in Qianbei Ma Goat (Capra hircus) and Its Association Analysis with Litter Size
ZHANG Yan1,2,3, CHEN Xiang1,2,3,*, ZHOU Zhi-Nan1,2,3, FU Kai-Bin1,2,3, WANG Zhong1,2,3, LI Shi-Jun1,2,3, YANG Pei-Fang1,2,3, HUI Mao-Mao1,2,3
1 Key Laboratory of Genetic Breeding and Breeding of Highland Mountain Animals, Ministry of Education, Guizhou University, Guiyang 550025, China; 2 Key Laboratory of Genetic Breeding and Breeding of Animals, Guizhou Province, Guiyang 550025, China; 3 School of Animal Science, Guizhou University, Guiyang 550025, China
Abstract：CytochromeP450c17 (CYP17A1) and cytochromeP450 aromatase (CYP19A1) were important for follicle development, ovulation and the secretion and synthesis of major reproductive hormones, and affected the reproductive performance of animals. In this study, healthy Qianbei Ma goat (Capra hircus) were taken as the research object. The tissue expression of CYP17A1 and CYP19A1 genes was detected by qPCR, and the SNP mutation sites were screened by PCR direct sequencing. The effect of SNPs on the lambing number of different parity of Qianbei Ma goat was analyzed by establishing a general linear model. The expression of CYP17A1 and CYP19A1 genes in various tissues and the correlation between SNPs and litter size were analyzed. The results showed that the expression of CYP17A1 gene was the highest in the ovary of single and multiple lambs, and was extremely significantly higher than that of pituitary, hypothalamus, uterus and fallopian tube (P<0.01). CYP19A1 gene expression was higher in the multi-lamb ovary and uterus, and both were higher than the corresponding single-lamb tissue (P<0.01). A total of 7 SNPs were detected in the CYP17A1 gene coded region. g.59G>C, g.128G>A, g.129A>G and g.164T>C were all located in the first exon, g.2045G>A was in the second exon and g.4387G>A and g.4456C>T were in the sixth exon. Correlation analysis showed that the SNPs of CYP17A1 gene were not correlated with the litter size of Qianbei Ma goat. A mutation site g.48818C>A was detected in the 9th exon of the coding region of CYP19A1 gene and the number of lambs in the AA genotype was significantly higher than that of the CA genotype (P<0.05), but not significantly different from that of the CC genotype. The results showed that the expression of CYP17A1 and CYP19A1 genes in Qianbei Ma goat was the highest in the ovarian tissue of multiple lambs (P<0.01). There were only 4 goats in 150 with AA genotype g.48818C>A site, which was not representative and needs to be verified. This study provides basic data for further exploring the relationship between CYP17A1 and CYP19A1 gene variants and animal reproduction.
张艳, 陈祥, 周志楠, 付开斌, 王忠, 李世军, 杨沛方, 惠茂茂. 黔北麻羊CYP17A1、CYP19A1基因组织表达、功能预测及与产羔数的关联性分析[J]. 农业生物技术学报, 2022, 30(6): 1140-1152.
ZHANG Yan, CHEN Xiang, ZHOU Zhi-Nan, FU Kai-Bin, WANG Zhong, LI Shi-Jun, YANG Pei-Fang, HUI Mao-Mao. Tissue Expression, Functional Prediction of CYP17A1 and CYP19A1 Genes in Qianbei Ma Goat (Capra hircus) and Its Association Analysis with Litter Size. 农业生物技术学报, 2022, 30(6): 1140-1152.
[1] 敖叶, 陈祥, 周志楠, 等. 2020. TGFβ2和PPP3CA基因在黔北麻羊性腺轴组织表达及对卵巢颗粒细胞的调控研究[J]. 畜牧兽医学报, 51(11): 2699-2709. (Ao Y, Chen X, Zhou Z N, et al.2020. TGFβ2 and PPP3CA gene expression in gonadal axis tissue and regulation of ovarian granulosa cells in Qianbei Ma goat[J]. Journal of Animal Husbandry and Veterinary Medicine, 51(11): 2699-2709.) [2] 李青, 卢曾奎, 金美林, 等 . 2019. 绵羊BMP2基因型与尾性状的关联分析[J]. 农业生物技术学报, 27(11):1985-1995. (Li Q, Lu Z K, Jin M L, et al.2019. Associationanalysis of the BMP2 genotype with sheep (Ovis aries)tail traits[J]. Journal of Agricultural Biotechnology, 27(11): 1985-1995.) [3] 邱星辉, 冷欣夫. 2000. 棉铃虫幼虫加单氧酶活性的组织分布[J]. 生态学报, 2(02): 299-203. (Qiu X H, Leng X F.2000. Tissue distribution of larval plus monooxygenase activity in cotton bollworm[J]. Journal of Ecology, 2(02): 299-203.) [4] 廖朝美, 游敏芳, 谭光辉, 等. 2021. 三穗鸭SCNN1B基因组织表达及其SNPs与蛋壳品质的相关性分析[J].农业生物技术学报, 29(04): 723-731. (Liao C M, You M F, Tan G H, et al.2021. Tissue expression of SCNN1B gene and correlation between its SNPs and egg shell quality in Sansui ducks[J]. Journal of Agricultural Biotechnology, 29(04): 723-731.) [5] 宋展, 高鑫, 吴冕, 等. 2020. 细胞色素P450酶的结构、功能与应用研究进展[J]. 微生物学通报, 47(07): 2245-2254. (Song Z, Gao X, Wu M, et al.2020. Advances in the structure, function and application of cytochrome P450 enzymes[J]. Microbiology Bulletin , 47(07): 2245-2254.) [6] 孙莹, 王萌, 孙晓军, 等. 2021. 牦牛卵泡发育及卵母细胞成熟过程中CYP19A1表达差异分析[J].甘肃畜牧兽医, 51(05): 37-42. (Sun Y, Wang M, Sun X J, et al.2021. Differential expression of CYP19A1 during follicular development and oocyte maturation in yak (Bos grunniens)[J]. Gansu Animal Husbandry and Veterinary Medicine,51(05): 37-42.) [7] 苏节, 刘庆友, 朱鹏, 等. 2013. 沼泽型水牛CYP19A1基因克隆、序列分析及组织表达研究[J].畜牧兽医学报, 44(04):514-521. (Su J, Liu Q Y, Zhu P, et al.2013. Cloning, sequence analysis and tissue expression of CYP19A1 gene in marsh-type buffalo[J]. Journal of Animal Husbandry and Veterinary Medicine,44(04):514-521.) [8] 唐振华. 1990. 细胞色素P450在昆虫适应性和抗药性中的作用[J]. 昆虫知识, 27(03): 175-178. (Tang Z H.1990. The role of cytochrome P450 in insect adaptation and drug resistance[J]. Insect Knowledge, 27(03): 175-178.) [9] 田志龙, 刘秋月, 王翔宇, 等. 2019. CYP11A1、CYP17A1、CYP19A1基因在小尾寒羊下丘脑-垂体-卵巢轴的表达[J]. 中国农业大学学报, 24(04): 87-93. (Tian Z L, Liu Q Y, Wang X Y, et al.2019. Expression of CYP11A1, CYP17A1 and CYP19A1 genes in the hypothalamic-pituitary-ovarian axis of the little tail cold sheep[J]. Journal of China Agricultural University, 24(04): 87-93.) [10] 田志龙. 2019. 绵羊繁殖候选基因的组织表达、SNP及其与产羔数的关联分析[D].硕士学位论文, 河南科技大学.导师: 王玉琴, pp. 4-8. (Tissue expression of candidate genes for reproduction, SNPs and their association with lambing number in sheep[D]. Thesis for M.S., Henan University of Science and Technology, Supervisor: Wang Y Q, pp. 4-8.) [11] 毛圆辉. 2015. 翻译过程中mRNA二级结构的功能研究[D]. 博士学位论文, 西北农林科技大学, 导师: 陶士珩, pp.6-7. (Mao Y H.2015. Functional study on the secondary structure of mRNA during translation [D]. Thesis for Ph.D., Northwest A&F University, Supervisor: Tao S H, pp. 6-7.) [12] 贾美婷, 李显耀, 康丽, 等. 2016. 寿光鸡GPR147基因5'调控区多态性与产蛋性状的关联分析[J].畜牧兽医学报, 47(01): 34-40. (Jia M T, Li X Y, Kang L, et al.2016. Association analysis of polymorphisms in the 5' regulatory region of the GPR147 gene with egg-laying traits in Shouguang chickens[J]. Journal of Animal Husbandry and Veterinary Medicine, 47(01): 34-40.) [13] 谭光辉, 吴磊, 李杰章, 等. 2020. 三穗鸭ITPR2基因组织表达、功能预测及其SNPs对蛋壳品质的影响[J]. 农业生物技术学报, 28(04): 720-730. (Tan G H, Wu L, Li J Z,et al.2020. Tissue expression and functional prediction of ITPR2 gene and the effect of its SNPs on eggshell quality in Sansui ducks[J]. Journal of Agricultural Biotechnology, 28(04): 720-730.) [14] 杨沛方, 周志楠, 敖叶, 等. 2020. 黔北麻羊StAR基因的克隆、生物信息学分析及在单、多羔黔北麻羊不同性腺组织中的表达[J]. 中国畜牧杂志, 56(11): 78-84. (Yang P F, Zhou Z N, Ao Y, et al.2020. Cloning, bioinformatics analysis and expression of StAR gene in different gonadal tissues of single and multi-lambed Qianbei ma goat[J]. Chinese Journal of Animal Husbandry, 56(11): 78-84.) [15] 周志楠, 陈祥, 张艳, 等. 2021. CTSD对黔北麻羊卵泡颗粒细胞的调控机制及功能分析[J].畜牧兽医学报, 52(05):1278-1292. (Zhou Z N,Chen X, Zhang Y, et al.2021. The regulatory mechanism and function analysis of CTSD on the granulosa cells of Qianbei Ma goat[J]. Journal of Animal Husbandry and Veterinary Medicine, 52(05): 1278-1292.) [16] 张辉, 张清阳, 李晨晨, 等. 2020. 太行鸡CYP19A1基因克隆及生物信息学分析[J]. 中国畜牧兽医, 47(05): 1489-1497. (Zhang H, Zhang Q Y, Li C C, et al.2020. Cloning and bioinformatics analysis of CYP19A1 gene in Taihang chickens[J]. China Veterinary Animal Husbandry, 47(05): 1489-1497.) [17] 张福平, 唐靓婷, 王嘉福, 等. 2021. 基于转录组测序筛选影响从江香猪产仔数的候选基因[J]. 南方农业学报, 52(04): 847-856. (Zhang F P, Tang L T, Wang J F, et al.2021. Transcriptome sequencing-based screening of candidate genes affecting litter size in Conjiang aromatic pigs[J]. Southern Journal of Agriculture, 52(04): 847-856.) [18] Chen M H, Li T, Ding C H, et al.2013. Growth differential factor-9 inhibits testosterone production in mouse theca interstitial cells[J]. Fertility and Sterility, 100(5): 1444-1450. [19] El-Bayomi K M, Saleh A A, Awad A, et al.2018. Association of CYP19A1 gene polymorphisms with anoestrus in water buffaloes[J]. Reproduction, Fertility and Development, 30(3): 487-497. [20] Gervásio C G, Bernuci M P, Silva-de-Sá M F, et al.2014. The role of androgen hormones in early follicular development[J]. Obstetrics and Gynecology, 2014(18): 81801. [21] Hess M F, Roser J F.2004. Immunocytochemical localization of cytochrome P450 aromatase in the testis of prepubertal, pubertal, and postpubertal horses[J]. Theriogenology, 61(2-3): 229-239. [22] Juengel J L, Hudson N L, Heath D A, et al.2002. Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep[J]. Biology of Reproduction, 67(06): 1777-1789 [23] Kwon J Y, McAndrew B J, Penman D J.2001. Cloning of brain aromatase gene and expression of brain and ovarian aromatase genes during sexual differentiation in genetic male and female Nile tilapia Oreochromis niloticus[J]. Molecular Reproduction and Development, 59(04):359-700. [24] Luo M, Li L, Xiao C, et al.2016. Heat stress impairs mice granulosa cell function by diminishing steroids production and inducing apoptosis[J]. Molecular and Cellular Biochemistry, 412(1-2): 81-90. [25] Padmanabhan V, Salvetti N R, Matiller V, at al.2014. Developmental programming: Prenatal steroid excess disrupts key members of intraovarian steroidogenic pathway in sheep[J]. Endocrinology, 155(9): 3649-3660. [26] Qiu X, Xiao X, Li N, et al.2019. Association of steroid 17-alpha-hydroxylase/17,20 lyase (CYP17) single nucleotide polymorphism (SNP) 628 and dopamine receptor D2(DRD2) SNP939 genotypes with sheep reproductive performance[J]. Journal of reproduction and fertility, 31(04): 743-750.) [27] Santos C P, Aguiar A F, Giom etti I C, et al.2018. High final energy of gallium arsenide laser increases MyoD gene expression during the intermediate phase of muscle regeneration after cryoinjury in rats[J]. Lasers in Medical Science, 33(04): 1-8. [28] Vega W H O, Quirino C R, Bartholazzi-Junior A, et al.2018. Variants in the CYP19A1 gene can affect in vitro embryo production traits in cattle[J]. Journal of Assisted Reproduction and Genetics, 35(12): 2233-2241.) [29] Wang J, Gong Y.2017.Transcription of CYP19A1 is directly regulated by SF-1 in the theca cells of ovary follicles in chicken[J]. General & Comparative Endocrinology, 247(01): 1-7. [30] Yuan J H, Wang J Z, Lan G C, et al.2008. Expression of steroidogenic enzymes and synthesis of steroid hormones during development of ovarian follicles in prepubertal goats[J]. Domest Anim Endocrinol, 34(4): 451-460. [31] Young J M, McNeilly A S.2012. Inhibin removes the inhibitory effects of activin on steroid enzyme expression and androgen production by normal ovarian thecal cells[J]. Journal of Molecular Endocrinology, 48(01): 49-60. [32] Zhang W, Lu H, Jiang H, et al.2012. Isolation and characterization of cyp19a1a and cyp19a1b promoters in the protogynous hermaphrodite orange-spotted grouper (Epinephelus coioides)[J]. General & Comparative Endocrinology, 175(03): 473-487. [33] Zhou R, Yang Y L, Liu Y, et al.2017. Association of CYP19A1 gene polymorphisms with reproductive traits in pigs[J]. Journal of Integrative Agriculture, 16(07):1558-1565.
