TGF-β RⅠ及Smad2表达情况对绵羊腔前卵泡发育的影响

常迪,董晓晨,郭勇,刘云海,齐晓龙,盛熙晖,盛熙辉,倪和民

中国农学通报. 2016, 32(32): 7-14

PDF(6544 KB)
PDF(6544 KB)
中国农学通报 ›› 2016, Vol. 32 ›› Issue (32) : 7-14. DOI: 10.11924/j.issn.1000-6850.casb16040076
畜牧 动物医学 蚕 蜂

TGF-β RⅠ及Smad2表达情况对绵羊腔前卵泡发育的影响

  • 常迪,董晓晨,郭勇,刘云海,齐晓龙,盛熙晖,倪和民
作者信息 +

Expression of TGF-β RⅠ and Smad2 Affecting the Development of Sheep Preantral Follicles

  • Chang Di, Dong Xiaochen, Guo Yong, Liu Yunhai, Qi Xiaolong, Sheng Xihui, Ni Hemin
Author information +
History +

摘要

为探索适于哺乳动物腔前卵泡的培养方式以及观察颗粒细胞与卵母细胞的信号表达，为阐明早期卵泡生长的复杂机理和揭示卵泡发生发育的内在规律提供参考，通过使用免疫组织化学染色、荧光免疫组织化学染色以及RT-PCR的方法，观察TGF-β RⅠ与Smad2的蛋白和mRNA在绵羊卵巢卵泡的表达情况。结果表明：TGF-β RⅠ在绵羊腔前卵泡及小腔卵泡的颗粒细胞及卵母细胞有表达，随着卵泡的生长发育，到成熟卵泡时期时，信号消失；Smad2蛋白在绵羊卵泡各个发育时期的颗粒细胞中均有表达，在绵羊腔前卵泡的卵母细胞中无表达，随着卵泡的生长发育，信号逐渐出现，到成熟卵泡时期时，Smad2蛋白在卵母细胞表达较强；TGF-β RⅠ mRNA只在成熟卵泡和培养前腔前卵泡颗粒细胞中检测到表达，在成熟卵泡和培养前腔前卵泡颗粒细胞中均有Smad2 mRNA表达，但在培养后腔前卵泡颗粒细胞中Smad2 mRNA表达量较低。TGF-β RⅠ与Smad2的蛋白和mRNA表达与绵羊卵泡发育情况密切相关。

Abstract

The aims of this study are to investigate the optimal way for cultivation of preantral follicles and observe the signal expression of granular cells and oocytes so as to explore the complex mechanism and inner law of folliculogenesis. The expressions of protein and mRNA of TGF-β RⅠ and Smad2 in sheep follicles were observed by using immunohistochemical staining, fluorescence immunohistochemical staining and RT-PCR. The results showed that TGF-β RⅠ signal expressed in granular cells and oocytes of sheep preantral follicles and small antral follicles but disappeared in mature follicles. Smad2 protein expressed in granular cells during all the development periods of sheep follicle, but did not express in oocytes of preantral follicles; with the development of follicle, the signal appeared and had a strong expression in oocytes of mature follicles. mRNA of TGF-β RⅠ only expressed in mature follicles and preantral follicles without culturing. mRNA of Smad2 expressed in all the follicles, but had a lower expression level in preantral follicles that were cultured for 6 days. The expression of protein and mRNA of TGF-β RⅠ and Smad2 have a close relation with the development of sheep follicles.

关键词

TGF-β RⅠ及Smad2表达情况对绵羊腔前卵泡发育的影响

Key words

TGF-β RⅠ; Smad2; immunohistochemistry; RT-PCR; follicle

引用本文

EndNote

Ris (Procite)

Bibtex

导出引用
常迪,董晓晨,郭勇,刘云海,齐晓龙,盛熙晖,盛熙辉,倪和民. TGF-β RⅠ及Smad2表达情况对绵羊腔前卵泡发育的影响. 中国农学通报. 2016, 32(32): 7-14 https://doi.org/10.11924/j.issn.1000-6850.casb16040076
Chang Di,Dong Xiaochen,Guo Yong,Liu Yunhai,Qi Xiaolong,Sheng Xihui, and Ni Hemin. Expression of TGF-β RⅠ and Smad2 Affecting the Development of Sheep Preantral Follicles. Chinese Agricultural Science Bulletin. 2016, 32(32): 7-14 https://doi.org/10.11924/j.issn.1000-6850.casb16040076

参考文献

[1] Nayudu P L, Fehrenbach A, Kiesel P, et al. Progress toward understanding follicle development in vitro: appearances are not deceiving[J].Arch Med Res 2001;32:587-594.
[2] Senbon S, Hirao Y, Miyano T. Interactions between the oocyte and surrounding somatic cells in follicular development: lessons from in vitro culture[J].J Reprod Dev,2003,49:259-269.
[3] Abir R, Nitke S, Ben-Haroush A, et al. In vitro maturation of human primordial ovarian follicles: clinical significance, progress in mammals, and methods for growth evaluation[J].Histol Histopathol 2006;21:887–898.
[4] Thomas F H, Walters K A, Telfer E E. How to make a good oocyte: an update on in-vitro models to study follicle regulation[J].Hum Reprod Update 2003,9:541-555.
[5] Demeestere I, Centner J, Gervy C, et al. Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents[J].Reproduction,2005,130:147-156.
[6] Ksiazkiewicz L K. Recent achievements in vitro culture and preservation of ovarian follicles in mammals[J].Reprod Biol,2006,6:3-16.
[7] Krieger P K, Deck J W, Woodruff T K, et al. The in vitro regulation of ovarian follicle development using alginate-extracellular matrix gels[J].Biomaterials,2006,27:714-723.
[8] Xu M, Krieger P K, Shea L D, et al. Tissue engineered follicles produce live, fertile offspring[J].Tissue Eng.2006,12(10):2739-2746.
[9] Xu M, West E, Shea L D, et al. Identification of a stage-specific permissive in vitro culture environment for follicle growth and oocyte development[J].Biol Reprod,2006,75:916-923.
[10] Somen Saha, Lin Ji, Juan J. de Pablo, and Sean P. Palecek. TGF-β/Activin/Nodal Pathway in Inhibition of Human Embryonic Stem Cell Differentiation by Mechanical Strain[J].Biophysical Journal,2008,98:4123-4133.
[11] Massague J, Wotton D. Transcriptional control by the TGF-beta/Smad signaling system[J].EMBO Journal,2000,19:1745-1754.
[12] Jerome Collignon. Activin/Nodal signalling before implantation: setting the stage for embryo natternina[J].Phil Trans R Soc,2014,369:1-8.
[13] 张超,罗艳梅,张家骅,等.激活素、抑制素及其受体与动物生殖作用的研究进展[J].中国畜牧兽医,2011,38(2):115-118.
[14] 罗艳梅,于明举,张超,等.抑制素-激活素-卵泡抑素系统及其受体的研究进展[J].黑龙江畜牧兽医,2011(9):22-23.
[15] Diana M H, Joanne C, Peter Bayliss, et al. Inhibitor-resistant type I receptors reveal specific requirements for TGF-β signaling in vivo[J].Developmental Biology,2006,295:730-742.
[16] 刘银,赵琴平,董惠芬,等.TGF-beta信号传导通路及其生物学功能[J].中国病原生物学杂志,2014,9(1):77-83.
[17] Gareth J Inman, Francisco J Nicola′ s, Caroline S Hill. Nucleocytoplasmic Shuttling of Smads 2, 3, and 4 Permits Sensing of TGF-β Receptor Activity[J].Molecular Cell,2002,10:283-294.
[18] 徐梦思.TGF-SMAD信号通路对猪颗粒细胞和繁殖性状的作用研究[D].石河子:石河子大学,2015.
[19] 曹雪梅,李武修,高玉光,等.TGF-β/Smad信号转导通路及其调控机制的研究进展[J].国际免疫学杂志,2008,31(6):482-488.
[20] Dai F, Lin X, Chang C, et al. Nuclear Export of Smad2 and Smad3 by RanBP3 Facilitates Termination of TGF-β Signaling[J].Developmental Cell,2009,16:345-357.
[21] Shiona M Coutts, Andrew J Childs, Norma Fulton, et al. Activin signals via SMAD2/3 between germ and somatic cells in the human fetal ovary and regulates kit ligand expression[J].Developmental Biology,2008,314:189-199.
[22] David Wotton, Roger S Lo, Susan Lee, et al. A Smad Transcriptional Corepressor[J].Cell,1999,97:29-39.
[23] Morgan Huse, Tom W Muir, Lan Xu, et al. The TGF-β Receptor Activation Process: An Inhibitor- to Substrate-Binding Switch[J].Molecular Cell,2001,8:671-682.
[24] Shimasaki S, Zachow R J, Li D, et al. A functional bone morphogenetic protein system in the ovary[J].PNAS,1999,96:7282-7287.
[25] Drummond A E, Dyson M, Le M T, et al. Ovarian follicle populations of the rat express TGF-beta signalling pathways[J].Molecular and Cellular Endocrinology,2003,202:53-57.
[26] Erickson G F, Shimasaki S. The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle[J].Reproductive Biology and Endocrinology,2003(1):9.
[27] Bristol S K, Woodruff T K. Follicle-restricted compartmentalization of transforming growth factor beta superfamily ligands in the feline ovary[J].Biology of Reproduction,2004,70:846-859.
[28] McNatty K P, Smith P, Moore L G, et al. Oocyte-expressed genes affecting ovulation rate[J].Molecular and Cellular Endocrinology,2005,234:57-66.
[29] 郭晶,李新宇,李隐侠,等.湖羊TGF-beta1基因特征、表达及其与排卵数的相关性分析[J].中国农业科学,2013,46(21):4586-4593.
[30] 汪正品.转化生长因子-beta参与调控小鼠卵巢原始卵泡库维持与激活的研究[D].北京:中国农业大学,2014:2-27.
[31] Dong C, Li Z, Rene Alvarez, et al. Microtubule Binding to Smads May Regulate TGF-β Activity Zhiru[J].Molecular Cell,2000(5):27-34.
[32] 郭永红,罗金燕.TGF-β超家族与Smad信号转导研究进展[J].医学综述,2005,11(8):685-672.
PDF(6544 KB)

Accesses

Citation

Detail
段落导航
相关文章

/

〈 〉