Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation

doi:10.1016/S1671-2927(00)8663

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (8): 1333-1339.DOI: 10.1016/S1671-2927(00)8663

Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation

ZHAO Xi-an, CANG Ming, GAO Xiao-yu, YANG Mei-ling, YUAN Jian-long, ZHU Bing, WANG Zhi-gang

Key Laboratory of Ministry of Education of China for Mammal Reproduction Biology and Biotechnology, Inner Mongolia University,Hohhot 010021, P.R.China

收稿日期:2011-05-20 出版日期:2012-08-01 发布日期:2012-09-09
通讯作者: Correspondence LIU Dong-jun, Tel/Fax: +86-471-4995071, E-mail: nmliudongjun@sina.com; CANG Ming, E-mail:cangming1970@yahoo.com.cn
作者简介:ZHAO Xi-an, E-mail:zxa_81@163.com;
基金资助:
The present study was supported by funding from the National 863 Program of China (2008AA1010070) and the Key Projects of the Natural Science Foundation of Inner Mongolia of China (20080404ZD08).

Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation

ZHAO Xi-an, CANG Ming, GAO Xiao-yu, YANG Mei-ling, YUAN Jian-long, ZHU Bing, WANG Zhi-gang

Key Laboratory of Ministry of Education of China for Mammal Reproduction Biology and Biotechnology, Inner Mongolia University,Hohhot 010021, P.R.China

Received:2011-05-20 Online:2012-08-01 Published:2012-09-09
Contact: Correspondence LIU Dong-jun, Tel/Fax: +86-471-4995071, E-mail: nmliudongjun@sina.com; CANG Ming, E-mail:cangming1970@yahoo.com.cn
About author:ZHAO Xi-an, E-mail:zxa_81@163.com;
Supported by:
The present study was supported by funding from the National 863 Program of China (2008AA1010070) and the Key Projects of the Natural Science Foundation of Inner Mongolia of China (20080404ZD08).

摘要/Abstract

摘要： To study the effects of interleukin-6 (IL-6) and its receptor (IL-6R) during in vitro maturation of ovine oocytes, the mRNA and protein expression levels of IL-6 and IL-6R, along with their localization, were examined during ovine oocytes maturation in vitro through real-time PCR, Western blotting, and immunohistochemistry. Specific patterns of expression of IL-6 and IL-6R were observed at both mRNA and protein levels at each stage of ovine oocytes maturation. IL-6 and IL-6R were distributed primarily on the surface of the cell membrane, with little expression in the cytoplasm or nucleus. IL-6 and IL- 6R were expressed significantly at higher levels in the maturation around 4 h, and then decreased dramatically. However the level slightly elevated at 20-24 h. The role of IL-6 and IL-6R on oocytes maturation was studied through in vitro addition of recombinant human IL-6 in different concentrations. The addition of 10 ng mL-1 IL-6 significantly increased the rates of oocytes maturation (P<0.05), but did not affect the rates of development of the subsequence IVF ovine embryos. In summary, IL-6 is likely to play an important role in the early ovine oocytes maturation. The expression patterns of the IL-6 and IL-6R on the ovine oocytes maturation open up the possibility of regulatory role of the cytokine in ovine oocytes maturation.

关键词: IL-6, oocyte, in vitro maturation, real time PCR, Western blotting

Abstract: To study the effects of interleukin-6 (IL-6) and its receptor (IL-6R) during in vitro maturation of ovine oocytes, the mRNA and protein expression levels of IL-6 and IL-6R, along with their localization, were examined during ovine oocytes maturation in vitro through real-time PCR, Western blotting, and immunohistochemistry. Specific patterns of expression of IL-6 and IL-6R were observed at both mRNA and protein levels at each stage of ovine oocytes maturation. IL-6 and IL-6R were distributed primarily on the surface of the cell membrane, with little expression in the cytoplasm or nucleus. IL-6 and IL- 6R were expressed significantly at higher levels in the maturation around 4 h, and then decreased dramatically. However the level slightly elevated at 20-24 h. The role of IL-6 and IL-6R on oocytes maturation was studied through in vitro addition of recombinant human IL-6 in different concentrations. The addition of 10 ng mL-1 IL-6 significantly increased the rates of oocytes maturation (P<0.05), but did not affect the rates of development of the subsequence IVF ovine embryos. In summary, IL-6 is likely to play an important role in the early ovine oocytes maturation. The expression patterns of the IL-6 and IL-6R on the ovine oocytes maturation open up the possibility of regulatory role of the cytokine in ovine oocytes maturation.

Key words: IL-6, oocyte, in vitro maturation, real time PCR, Western blotting

ZHAO Xi-an, CANG Ming, GAO Xiao-yu, YANG Mei-ling, YUAN Jian-long, ZHU Bing, WANG Zhi-gang. Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation[J]. Journal of Integrative Agriculture, 2012, 12(8): 1333-1339.

参考文献

[1]Asimakopoulos B, Nikolettos N, Papachristou D N, Simopoulou M, Al-hasani S, Diedrich K. 2005. Follicular fluid levels of vascular endothelial growth factor and leptin are associated with pregnancy outcome of normal women participating in intracytoplasmic sperm injection cycles. Physiological Research, 54, 263-270.

[2]Byskov A G, Andersen C Y, Leonardsen L. 2002. Role of meiosis activating sterols, MAS, in induced oocyte maturation. Molecular and Cellular Endocrinology, 187, 189-196.

[3]Chen S U, Chou C H, Lee H, Ho C H, Lin C W, Yang Y S. 2008. Lysophosphatidic acid up-regulates expression of interleukin-8 and-6 in granulosa-lutein cells through its receptors and nuclear factor-kappaB dependent pathways: implications for angiogenesis of corpus luteum and ovarian hyperstimulation syndrome. Clinical Endocrinology and Metabolism, 93, 935-943.

[4]Chian R C, Lim J H, Tan S L. 2004. State of the art in in-vitro oocyte maturation. Current Opinion in Obstetrics and Gynecology, 16, 211-219.

[5]Gordo A C, He C L, Smith S, Fissore R A. 2001. Mitogen activated protein kinase plays a significant role in metaphase II arrest, spindle morphology, and maintenance of maturation promoting factor activity in bovine oocytes. Molecular Reproduction and Development, 59, 106-114.

[6]Grøndahl C. 2008. Oocyte maturation. Basic and clinical aspects of in vitro maturation (IVM) with special emphasis of the role of FF-MAS. Danish Medical Bulletin, 55, 1-16.

[7]Heinrich P C, Behrmann I, Haan S, Hermanns H M, Müller-Newen G, Schaper F. 2003. Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochemical Journal, 374, 1-20.

[8]Kawano Y, Kawasaki F, Nakamura S, Matsui N, Narahara H, Miyakawa I. 2001. The production and clinical evaluation of macrophage colony-stimulating factor and macrophage chemoattractant protein-1 in human follicular fluid. American Journal of Reproductive Immunology, 45, 1-5.

[9]Kawasaki F, Kawano Y, Kosay Hasan Z, Narahara H, Miyakawa I. 2003. The clinical role of interleukin-6 and interleukin-6 soluble receptor in human follicular fluids. Clinical and Experimental Medicine, 3, 27-31.

[10]Kubelka M, Motlik J, Fulka J J, Procházka R, Rimkevicová Z, Fulka J. 1988. Time sequence of germinal vesicle breakdown in pig oocytes after cycloheximide and paminobenzamidine block. Gamete Research, 19, 423-431.

[11]Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C (T) method. Methods, 25, 402-408.

[12]Maeda A, Inoue N, Matsuda-Minehata F, Goto Y, Cheng Y, Manabe N. 2007. The role of interleukin-6 in the regulation of granulosa cell apoptosisduring follicular atresia in pig ovaries. Journal of Reproduction and Development, 53, 481-490.

[13]Ptak G, Lopes F, Matsukawa K, Tischner M, Loi P. 2006. Leukaemia inhibitory factor enhances sheep fertilization i n v i t r o v i a a n i n f l u e n c e o n t h e o o c y t e . Theriogenology, 65, 1891-1899.

[14]Rose-John S, Scheller J, Elson G, Jones S A. 2006. Interleukin-6 biology is coordinated by membranebound and soluble receptors: role in inflammation and cancer. Journal of Leukocyte Biology, 80, 227-236.

[15]Rose-John S, Waetzig G H, Scheller J, Grötzinger J, Seegert D. 2007. The IL-6/sIL-6R complex as a novel target for therapeutic approaches. Expert Opinion on Therapeutic Targets, 11, 613-624.

[16]Sun Q Y, Lai L, Bonk A, Prather R S, Schatten H. 2001. Cytoplasmic changes in relation to nuclear maturation and early embryo developmental potential of porcine oocytes: effects of gonadotropins, cumulus cells, follicular size, and protein synthesis inhibition. Molecular Reproduction and Development, 59, 192-198.

[17]Tamura K, Kawaguchi T, Kogo H. 2001. Interleukin-6 inhibits the expression of luteinizing hormone receptor mRNA during the maturation of cultured rat granulosa cells. Endocrinology, 170, 121-127.

[18]Wang G H, Zhao H B, Xin X Y. 2003. Stem cell factor in human follicular fluid and its effects on the development of ovarian follicle. Journal of the Fourth Military Medical University, 24, 543-545.

[19]Watson A J. 2007. Oocyte cytoplasmic maturation: A key mediator of oocyte and embryo developmental competence. Journal of Animal Science, 85, E1-E3.

[20]Zhao H B, Liu Y. 2001. Concentrations of TNF-α and IL-6 in follicular fluid and their influence on ovarian function. Journal of the Fourth Military Medical University, 22, 429-431.

[21]Zhou H L, Ma J, Ji W Z. 2002. Developmental competence of ovine oocytes after in vitro inhibition of meiotic resumption. Acta Zoologica Sinica, 48, 741-748. (in Chinese)

[22]Zhou P, Liu D J, Yan Z, Ma Y Z, Bou S. 2007. The effects of TGFα and EGF on ovine oocytes maturation. Chinese Journal of Cell Biology, 29, 713-717. (in Chinese)

Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation

Expression of Interleukin-6 and Interleukin-6 Receptor in Ovine Oocytes During In vitro Maturation

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

[1]	JIANG Dai-xun, ZHANG Mei-hua, ZHANG Qian, CHEN Yi-shan, MA Wen-jing, WU Wei-peng, MU Xiang. Influence of gallic acid on porcine neutrophils phosphodiesterase 4, IL-6, TNF-α and rat arthritis model[J]. Journal of Integrative Agriculture, 2015, 14(4): 758-764.
[2]	XU Guang-yong, JIANG Jin-qi, WANG Ming, LI Jie, SU Jing-liang , REN Xiao-ming. Lactic Acid Reduces LPS-Induced TNF- and IL-6 mRNA Levels Through Decreasing I B Phosphorylation[J]. Journal of Integrative Agriculture, 2013, 12(6): 1073-1078.