Full Text:   <1220>

CLC number: Q93

On-line Access: 2010-09-08

Received: 2010-02-22

Revision Accepted: 2010-05-12

Crosschecked: 2010-08-09

Cited: 7

Clicked: 3573

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.9 P.719-727


Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells

Author(s):  Wei Wang, Li Wang, Xin-xiu Li, Xia Chen, Hai-yan Zhang, Yu He, Jing-jing Wang, Yong-yan Zhao, Bao-le Zhang, Yin-xue Xu

Affiliation(s):  College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Corresponding email(s):   xuyinxue@njau.edu.cn

Key Words:  Porcine granulosa cells, Bone morphogenetic protein (BMP)/Smad, Smad4, RNA interference (RNAi), Growth, Steroidogenesis

Wei Wang, Li Wang, Xin-xiu Li, Xia Chen, Hai-yan Zhang, Yu He, Jing-jing Wang, Yong-yan Zhao, Bao-le Zhang, Yin-xue Xu. Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells[J]. Journal of Zhejiang University Science B, 2010, 11(9): 719-727.

@article{title="Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells",
author="Wei Wang, Li Wang, Xin-xiu Li, Xia Chen, Hai-yan Zhang, Yu He, Jing-jing Wang, Yong-yan Zhao, Bao-le Zhang, Yin-xue Xu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells
%A Wei Wang
%A Li Wang
%A Xin-xiu Li
%A Xia Chen
%A Hai-yan Zhang
%A Yu He
%A Jing-jing Wang
%A Yong-yan Zhao
%A Bao-le Zhang
%A Yin-xue Xu
%J Journal of Zhejiang University SCIENCE B
%V 11
%N 9
%P 719-727
%@ 1673-1581
%D 2010
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000079

T1 - Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells
A1 - Wei Wang
A1 - Li Wang
A1 - Xin-xiu Li
A1 - Xia Chen
A1 - Hai-yan Zhang
A1 - Yu He
A1 - Jing-jing Wang
A1 - Yong-yan Zhao
A1 - Bao-le Zhang
A1 - Yin-xue Xu
J0 - Journal of Zhejiang University Science B
VL - 11
IS - 9
SP - 719
EP - 727
%@ 1673-1581
Y1 - 2010
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1000079

Bone morphogenetic proteins (BMPs) play a critical role in the growth and steroidogenesis of granulosa cells (GCs). BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors. Upon ligand binding, BMPs activate intracellular Smad proteins and regulate growth and apoptosis in various cell types. The objective of this study was to demonstrate the effects of BMP/Smad signal on growth and steroidogenesis of porcine GCs. A strategy of RNA interference (RNAi)-mediated ‘gene silencing’ of smad4, a core molecule mediating the intracellular BMP/Smad signal transduction pathways, was used to interrupt endogenous BMP/Smad signaling. Results indicate that smad4-small interfering RNA (siRNA) caused specific inhibition of smad4 mRNA and protein expression after transfection. Interrupted endogenous BMP/Smad signaling significantly inhibited growth, and induced apoptosis of porcine GCs, while decreasing estradiol production. In addition, interrupted BMP/Smad signaling significantly (P<0.05) changed the expression of Cyclin D2, CDK4, Bcl-2, and Cyp19a1. These findings provide new insights into how BMP/Smad signaling regulates the growth and steroidogenesis of porcine GCs.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Biondi, C.A., Das, D., Howell, M., Islam, A., Bikoff, E.K., Hill, C.S., Robertson, E.J., 2007. Mice develop normally in the absence of Smad4 nucleocytoplasmic shuttling. Biochem. J., 404(2):235-245.

[2]Costello, I., Biondi, C.A., Taylor, J.M., Bikoff, E.J., Robertson, E.J., 2009. Smad4-dependent pathways control basement membrane deposition and endodermal cell migration at early stages of mouse development. BMC Devel. Biol., 9(1):54.

[3]Dai, F., Lin, X., Chang, C., Feng, X.H., 2009. Nuclear export of Smad2 and Smad3 by RanBP3 facilitates termination of TGF-β signaling. Dev. Cell, 16(3):345-357.

[4]Dorsett, Y., Tuschl, T., 2004. siRNAs: applications in functional genomics and potential as therapeutics. Nat. Rev. Drug Discov., 3(4):318-329.

[5]Fan, H.Y., Shimada, M., Liu, Z.L., Cahill, N., Noma, N., Wu, Y., Gossen, J., Richards, J.S., 2008. Selective expression of KrasG12D in granulosa cells of the mouse ovary causes defects in follicle development and ovulation. Development, 135(12):2127-2137.

[6]Fan, H.Y., Liu, Z.L., Shimada, M., Sterneck, E., Johnson, P.F., Hedric, S.M., Richards, J.S., 2009. MAPK3/1 (ERK1/2) in ovarian granulosa cells are essential for female fertility. Science, 324(5929):938-941.

[7]Fraser, M., Braems, G.A., Challis, J.R., 2001. Developmental regulation of corticotrophin receptor gene expression in the adrenal gland of the ovine fetus and newborn lamb: effects of hypoxia during late pregnancy. J. Endocrinol., 169(1):1-10.

[8]Kawashima, I., Okazaki, T., Noma, N., Nishibori, M., Yamashita, Y., Shimada, M., 2008. Sequential exposure of porcine cumulus cells to FSH and/or LH is critical for appropriate expression of steroidogenic and ovulation-related genes that impact oocyte maturation in vivo and vitro. Reproduction, 136(1):9-21.

[9]Khamsi, F., Roberge, S., 2001. Granulosa cells of the cumulus oophorus are different from mural granulosa cells in their response to gonadotrophins and IGF-1. J. Endocrinol., 170(3):565-573.

[10]Lee, W.S., Otsuka, F., Moore, R.K., Shimasaki, S., 2001. Effect of bone morphogenetic protein-7 on folliculo-genesis and ovulation in the rat. Biol. Reprod., 65(4):994-999.

[11]Lin, X., Duan, X.Y., Liang, Y.Y., Su, Y., Wrighton, K.H., Long, J.Y., Hu, M., Davis, C.M., Wang, J.R., Brunicardi, F.C., et al., 2006. PPM1A functions as a Smad phosphatase to terminate TGFβ signaling. Cell, 125(5):915-928.

[12]Livak, K.J., Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆C<sub>T</sub> method. Methods, 25(4):402-408.

[13]Massagué, J., Seoane, J., Wotton, D., 2005. Smad transcription factors. Genes Dev., 19(23):2783-2810.

[14]Matsuura, I., Denissova, N.G., Wang, G., He, D.M., Long, J.Y., Liu, F., 2004. Cyclin-dependent kinases regulate the antiproliferative function of Smads. Nature, 430(6996):226-231.

[15]McGee, E.A., Hsueh, A.J.W., 2000. Initial and cyclic recruitment of ovarian follicles. Endocr. Rev., 21(2):200-214.

[16]Mittal, V., 2004. Improving the efficiency of RNA interference in mammals. Nat. Rev. Genet., 5(5):355-365.

[17]Miyoshi, T., Otsuka, F., Suzuki, J., Takeda, M., Inagaki, K., Kano, Y., Otani, H., Mimura, Y., Ogura, T., Makino, H., 2006. Mutual regulation of follicle-stimulating hormone signaling and bone morphogenetic protein system in human granulosa cells. Biol. Reprod., 74(6):1073-1082.

[18]Otsuka, F., Yao, Z.X., Lee, T.H., Yamamot, S., Erickson, G.F., Shimasaki, S., 2000. Bone morphogenetic protein-15. Identification of target cells and biological functions. J. Biol. Chem., 275(50):39523-39528.

[19]Otsuka, F., Moore, R.K., Shimasaki, S., 2001a. Biological function and cellular mechanism of bone morphogenetic protein-6 in the ovary. J. Biol. Chem., 276(35):32889-32895.

[20]Otsuka, F., Yamamoto, S., Erickson, G.F., Shimasaki, S., 2001b. Bone morphogenetic protein-15 inhibits follicle-stimulating hormone (FSH) action by suppressing FSH receptor expression. J. Biol. Chem., 276(14):11387-11392.

[21]Pangas, S.A., Li, X.H., Robertson, E.J., Matzuk, M.M., 2006. Premature luteinization and cumulus cell defects in ovarian-specific Smad4 knockout mice. Mol. Endocrinol., 20(6):1406-1422.

[22]Pangas, S.A., Li, X.H., Umans, L., Zwijsen, A., Huylebroeck, D., Gutierrez, C., Wang, D.G., Martin, J.F., Jamin, S.P., Behringer, R.R., et al., 2008. Conditional deletion of Smad1 and Smad5 in somatic cells of male and female gonads leads to metastatic tumor development in mice. Mol. Cell. Biol., 28(1):248-257.

[23]Paterson, I.C., Davies, M., Stone, A., Huntley, S., Smith, E., Pring, M., Eveson, J.W., Robinson, C.M., Parkinson, E.K., Prime, S.S., 2002. TGF-β1 acts as a tumor suppressor of human malignant keratinocytes independently of Smad4 expression and ligand-induced G(1) arrest. Oncogene, 21(10):1616-1624.

[24]Picton, H.M., Campbell, B.K., Hunter, M.G., 1999. Maintenance of oestradiol production and exression of cytochrome P450 aromatase enzyme mRNA in long-term serum-free cultures of pig granulosa cells. J. Reprod. Fertil., 115(1):67-77.

[25]Schmierer, B., Hill, C.S., 2007. TGFβ-SMAD signal transduction: molecular specificity and functional flexibility. Nat. Rev. Mol. Cell Biol., 8(12):970-982.

[26]Shimasaki, S., Zachow, R.J., Li, D.M., Kim, H., Iemura, S.I., Ueno, N., Sampath, K., Chang, R.J., Erickson, G.F., 1999. A functional bone morphogenetic protein system in the ovary. PNAS, 96(13):7282-7287.

[27]Shimasaki, S., Moore, R.K., Erickson, G.F., Otsuka, F., 2003. The role of bone morphogenetic proteins in ovarian function. Reprod. Suppl., 61:323-337.

[28]Shimasaki, S., Moore, R.K., Otsuka, F., Erickson, G.F., 2004. The bone morphogenetic protein system in mammalian reproduction. Endocr. Rev., 25(1):72-101.

[29]Vitt, U.A., Hayashi, M., Klein, C., Hsueh, A.J., 2000. Growth differentiation factor-9 stimulates proliferation but suppresses the follicle-stimulating hormone-induced differentiation of cultured granulosa cells from small antral and preovulatory rat follicles. Biol. Reprod., 62(2):370-377.

[30]Winters, T.A., Hanten, J.A., Veldhuis, J.D., 1998. In situ amplification of the cytochrome P-450 cholesterol side-chain cleavage enzyme mRNA in single porcine granulosa cells by IGF-1 and FSH acting alone or in concert. Endocrine, 9(1):57-63.

[31]Yamashita, Y., Kawashima, I., Yanai, Y., Nishibori, M., Tichards, J.S., Shimada, M., 2007. Hormone-induced expression of tumor necrosis factor α-converting enzyme/A disintegrin and metalloprotease-17 impacts porcine cumulus cell oocyte complex expansion and meiotic maturation via ligand activation of the epidermal growth factor receptor. Endocrinology, 148(12):6164-6175.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE