UBE2I stimulates female gonadal differentiation in chicken (Gallus gallus) embryos

doi:10.1016/S2095-3119(20)63486-4

Journal of Integrative Agriculture ›› 2021, Vol. 20 ›› Issue (11): 2986-2994.DOI: 10.1016/S2095-3119(20)63486-4

收稿日期:2020-05-09 出版日期:2021-11-01 发布日期:2021-09-17

UBE2I stimulates female gonadal differentiation in chicken (Gallus gallus) embryos

JIN Kai^{1, 2, 3*}, ZHOU Jing^{1, 2, 3*}, ZUO Qi-sheng^{1, 2, 3}, LI Jian-cheng^{1, 2, 3}, Jiuzhou SONG⁴, ZHANG Ya-ni^{1, 2, 3}, CHANG Guo-bing^{1, 2, 3}, CHEN Guo-hong^{1, 2, 3}, LI Bi-chun^{1, 2, 3}

¹ Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P.R.China
² Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, P.R.China
³ Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou 225009, P.R.China
⁴ Animal & Avian Sciences, University of Maryland, College Park, MD, 20741, USA

Received:2020-05-09 Online:2021-11-01 Published:2021-09-17
Contact: Correspondence LI Bi-chun, Tel: +86-514-87977207, Fax: +86-514-87350440, E-mail: yubcli@yzu.edu.cn
About author: JIN Kai, E-mail: jinkai0621@163.com; * These authors contributed equally to this study.
Supported by:
This work was funded by the National Natural Science Foundation of China (31772582 and 31972547), the National Key R&D Program of China (2017YFE0108000), the High Level Talents Support Program of Yangzhou University, Postgraduate Research & Practice Innovation Program of Jiangsu Province, China (KYCX18_2376), the Jiangsu Science and Technology Project, China (Youth Fund, BK20180918), and the Natural Science Research Project of Jiangsu Higher Education Institutions, China (18KJB230008).

摘要/Abstract

摘要：

该研究报道了一个新的雌性特异表达基因UBE2I，鉴定了其在鸡两性组织器官中的表达模式，并通过体内实验（干扰和过表达）研究了UBE2I在鸡胚性腺发育中的生物学功能。研究发现，UBE2I在鸡早期胚胎性腺中的PGCs（原始生殖细胞）中高表达，并在新生鸡卵巢组织中表现出高表达。与此同时，研究团队结合慢病毒和鸡胚血管注射技术建立了在鸡胚体内长期稳定干扰或过表达UBE2I表达的有效方法，并发现在UBE2I过表达的情况下，雌性的性别相关基因（FOXL2，CYP19A1和HINTW）被上调，而雄性的性别相关基因（SOX9，DMRT1和WT1）在干扰UBE2I后被下调，其对应的鸡胚性腺的组织也出现了对应的结构变化。与此同时，UBE2I表达的变化与雌二醇及其受体（AR和ESR）的水平有关，这些结果表明UBE2I是启动雌性鸡胚卵巢发育的关键因素。该研究结果有力的证明了UBE2I在鸡胚胎性别发育和分化过程中发挥了关键的作用。这是第一次报道UBE2I基因在鸡胚性别分化过程中发挥的功能，而UBE2I作为泛素化修饰过程中的重要调控因子，为进一步研究家禽性别决定的分子机理提供了新的思路。

Abstract:

Without known analogous sex-determining factors like SRY (sex determining region Y) in mammals, the chicken (Gallus gallus) sex determination mechanism still remains unclear, which highly restricts the biological research on chicken development and poultry single-sex reproduction. Here we not only characterized a new female-biased gene UBE2I and identified the expression pattern by qRT-PCR, but also described the functional role of UBE2I in the gonadal development of chicken embryos. Results showed that UBE2I exhibited a female-biased expression pattern in the early stage of PGCs (primordial germ cells) in embryonic gonads and robust expression in ovaries of newborn chickens. Most importantly, we successfully developed an effective method to interfere or overexpress UBE2I in chicken embryos through the intravascular injection. The qRT-PCR analysis showed that the sex-related genes (FOXL2, CYP19A1 and HINTW) in females were upregulated (P<0.05) under the overexpression of UBE2I and the sex-related genes (SOX9, DMRT1 and WT1) in females were downregulated (P<0.05) after interfering UBE2I. Furthermore, the change of UBE2I expression was associated with the level of estradiol and its receptors (AR and ESR), which suggests that UBE2I is necessary to initiate the female-specific development in chickens. In conclusion, this work demonstrates that UBE2I is a crucial sex differentiation-related gene in the embryonic development of chickens, which provides insights for further understanding the mechanism of sex determination in chickens.

Key words: UBE2I , gonadal differentiation , chicken embryonic , sex differentiation

. [J]. Journal of Integrative Agriculture, 2021, 20(11): 2986-2994.

JIN Kai, ZHOU Jing, ZUO Qi-sheng, LI Jian-cheng, Jiuzhou SONG, ZHANG Ya-ni, CHANG Guo-bing, CHEN Guo-hong, LI Bi-chun. UBE2I stimulates female gonadal differentiation in chicken (Gallus gallus) embryos[J]. Journal of Integrative Agriculture, 2021, 20(11): 2986-2994.

参考文献

Abinawanto, Shimada K, Yoshida K, Saito N. 1996. Effects of aromatase inhibitor on sex differentiation and levels of P450 (17alpha) and P450 arom messenger ribonucleic acid of gonads in chicken embryos. General and Comparative Endocrinology, 102, 241–246.
Castro J, Torres M, Sovino H, Fuentes A, Boric M A, Johnson M C. 2010. P450Arom induction in isolated control endometrial cells by peritoneal fluid from women with endometriosis. Fertility and Sterility, 94, 2521–2527.
Ellis H L, Shioda K, Rosenthal N F, Coser K R, Shioda T. 2012. Masculine epigenetic sex marks of the CYP19A1/aromatase promoter in genetically male chicken embryonic gonads are resistant to estrogen-induced phenotypic sex conversion. Biology of Reproduction, 87, 1–12, 23.
Hamburger V, Hamilton H L. 1951. A series of normal stages in the development of the chick embryo. Journal of Morphology, 88, 49–92.
He L, Martins P, Huguenin J, Van T N N, Manso T, Galindo T, Gregoire F, Catherinot L, Molina F, Espeut J. 2019. Simple, sensitive and robust chicken specific sexing assays, compliant with large scale analysis. PLoS ONE, 14, e0213033.
He X, Lai Q, Chen C, Li N, Sun F, Huang W, Zhang S, Yu Q, Yang P, Xiong F, Chen Z, Gong Q, Ren B, Weng J, Eizirik D L, Zhou Z, Wang C Y. 2018. Both conditional ablation and overexpression of E2 SUMO-conjugating enzyme (UBC9) in mouse pancreatic beta cells result in impaired beta cell function. Diabetologia, 61, 881–895.
Hirst C E, Major A T, Smith C A. 2018. Sex determination and gonadal sex differentiation in the chicken model. The International Journal of Developmental Biology, 62, 153–166.
Hudson Q J, Smith C A, Sinclair A H. 2005. Aromatase inhibition reduces expression of FOXL2 in the embryonic chicken ovary. Developmental Dynamics: An Official Publication of the American Association of Anatomists, 233, 1052–1055.
Ichikawa K, Ezaki R, Furusawa S, Horiuchi H. 2019. Comparison of sex determination mechanism of germ cells between birds and fish: Cloning and expression analyses of chicken forkhead box L3-like gene. Developmental Dynamics: An Official Publication of the American Association of Anatomists, 248, 826–836.
Ihara M, Stein P, Schultz R M. 2008. UBE2I (UBC9), a SUMO-conjugating enzyme, localizes to nuclear speckles and stimulates transcription in mouse oocytes. Biology of Reproduction, 79, 906–913.
Jin K, Chen H, Zuo Q, Huang C, Zhao R, Yu X, Wang Y, Zhang Y, Chang Z, Li B. 2018. CREPT and p15RS regulate cell proliferation and cycling in chicken DF-1 cells through the Wnt/β-catenin pathway. Journal of Cellular Biochemistry, 119, 1083–1092.
Kanai Y, Hiramatsu R, Matoba S, Kidokoro T. 2005. From SRY to SOX9: Mammalian testis differentiation. Journal of Biochemistry, 138, 13–19.
Kobayashi S, Shibata H, Kurihara I, Yokota K, Suda N, Saito I, Saruta T. 2004. Ubc9 interacts with chicken ovalbumin upstream promoter-transcription factor I and represses receptor-dependent transcription. Journal of Molecular Endocrinology, 32, 69–86.
Kurihara I, Shibata H, Kobayashi S, Suda N, Ikeda Y, Yokota K, Murai A, Saito I, Rainey W E, Saruta T. 2005. Ubc9 and protein inhibitor of activated STAT 1 activate chicken ovalbumin upstream promoter-transcription factor i-mediated human CYP11B2 gene transcription. The Journal of Biological Chemistry, 280, 6721–6730.
Lambeth L S, Cummins D M, Doran T J, Sinclair A H, Smith C A. 2013. Overexpression of aromatase alone is sufficient for ovarian development in genetically male chicken embryos. PLoS ONE, 8, e68362.
Lambeth L S, Morris K R, Wise T G, Cummins D M, O’Neil T E, Cao Y, Sinclair A H, Doran T J, Smith C A. 2016. Transgenic chickens overexpressing aromatase have high estrogen levels but maintain a predominantly male phenotype. Endocrinology, 157, 83–90.
Ma Z, Yu Y, Tang S, Liu H, Su G, Xie Y, Giesy J P, Hecker M, Yu H. 2015. Differential modulation of expression of nuclear receptor mediated genes by tris(2-butoxyethyl) phosphate (TBOEP) on early life stages of zebrafish (Danio rerio). Aquatic Toxicology (Amsterdam, Netherlands), 169, 196–203.
Rodriguez A, Briley S M, Patton B K, Tripurani S K, Rajapakshe K, Coarfa C, Rajkovic A, Andrieux A, Dejean A, Pangas S A. 2019. Loss of the E2 SUMO-conjugating enzyme Ube2i in oocytes during ovarian folliculogenesis causes infertility in mice. Development (Cambridge, England), 146, dev.176701.
Sechman A. 2013. The role of thyroid hormones in regulation of chicken ovarian steroidogenesis. General and Comparative Endocrinology, 190, 68–75.
Sekido R. 2010. SRY: A transcriptional activator of mammalian testis determination. The International Journal of Biochemistry & Cell Biology, 42, 417–420.
Smith C A, Sinclair A H. 2001. Sex determination in the chicken embryo. The Journal of Experimental Zoology, 290, 691–699.
Smith C A, Roeszler K N, Ohnesorg T, Cummins D M, Farlie P G, Doran T J, Sinclair A H. 2009a. The avian Z-linked gene DMRT1 is required for male sex determination in the chicken. Nature, 461, 267–271.
Smith C A, Roeszler K N, Sinclair A H. 2009b. Genetic evidence against a role for W-linked histidine triad nucleotide binding protein (HINTW) in avian sex determination. The International Journal of Developmental Biology, 53, 59–67.
Smith C A, Sinclair A H. 2004. Sex determination: Insights from the chicken. BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology, 26, 120–132.
Tang X, Li W, Xing J, Sheng X, Zhan W. 2016. SUMO and SUMO-conjugating enzyme E2 UBC9 are involved in white spot syndrome virus infection in fenneropenaeus chinensis. PLoS ONE, 11, e0150324.
Tizard M L, Jenkins K A, Cooper C A, Woodcock M E, Challagulla A, Doran T J. 2019. Potential benefits of gene editing for the future of poultry farming. Transgenic Research, 28, 87–92.
Tran H T, Ferrell W, Butt T R. 2010. An estrogen sensor for poultry sex sorting. Journal of Animal Science, 88, 1358–1364.
Western P S, Harry J L, Marshall G J A, Sinclair A H. 2000. Temperature-dependent sex determination in the American alligator: Expression of SF1, WT1 and DAX1 during gonadogenesis. Gene, 241, 223–232.
Yamashita S, Kataoka K, Yamamoto H, Kato T, Hara S, Yamaguchi K, Renard-Guillet C, Katou Y, Shirahige K, Ochi H, Ogino H, Uchida T, Inui M, Takada S, Shigenobu S, Asahara H. 2019. Comparative analysis demonstrates cell type-specific conservation of SOX9 targets between mouse and chicken. Scientific Reports, 9, 12560.
Yin Y, Kundu K, Pal L R, Moult J. 2017. Ensemble variant interpretation methods to predict enzyme activity and assign pathogenicity in the CAGI4 NAGLU (human N-acetyl-glucosaminidase) and UBE2I (human SUMO-ligase) challenges. Human Mutation, 38, 1109–1122.
Zhang J, Kinch L N, Cong Q, Weile J, Sun S, Cote A G, Roth F P, Grishin N V. 2017. Assessing predictions of fitness effects of missense mutations in SUMO-conjugating enzyme UBE2I. Human Mutation, 38, 1051–1063.
Zhang Z, Elsayed A K, Shi Q, Zhang Y, Zuo Q, Li D, Lian C, Tang B, Xiao T, Xu Q, Chang G, Chen G, Zhang L, Wang K, Wang Y, Jin K, Wang Y, Song J, Cui H, Li B. 2015. Crucial genes and pathways in chicken germ stem cell differentiation. Journal of Biological Chemistry, 290, 13605–13621.
Zuo Q, Jin K, Song J, Zhang Y, Chen G, Li B. 2018. Interaction of the primordial germ cell-specific protein C2EIP with PTCH2 directs differentiation of embryonic stem cells via HH signaling activation. Cell Death and Disease, 9, 497.

UBE2I stimulates female gonadal differentiation in chicken (Gallus gallus) embryos

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics