Oocyte-derived BMP15 but Not GDF9 Down-regulates Connexin43 Expression and Decreases Gap Junction Intercellular Communication Activity in Immortalized Human Granulosa Cells

Overview

Journal Mol Hum Reprod

Specialties Molecular Biology
Reproductive Medicine

Date 2014 Jan 14

PMID 24413384

Citations 22

Authors

Hsun-Ming Chang

Jung-Chien Cheng

Elizabeth Taylor

Peter C K Leung

Affiliations

Soon will be listed here.

Abstract

In the ovary, connexin-coupled gap junctions in granulosa cells play crucial roles in follicular and oocyte development as well as in corpus luteum formation. Our previous work has shown that theca cell-derived bone morphogenetic protein (BMP)4 and BMP7 decrease gap junction intercellular communication (GJIC) activity via the down-regulation of connexin43 (Cx43) expression in immortalized human granulosa cells. However, the effects of oocyte-derived growth factors on Cx43 expression remain to be elucidated. The present study was designed to investigate the effects of oocyte-derived growth differentiation factor (GDF)9 and BMP15 on the expression of Cx43 in a human granulosa cell line, SVOG. We also examined the effect relative to GJIC activity and investigated the potential mechanisms of action. In SVOG cells, treatment with BMP15 but not GDF9 significantly decreased Cx43 mRNA and protein levels and GJIC activity. These suppressive effects, along with the induction of Smad1/5/8 phosphorylation, were attenuated by co-treatment with a BMP type I receptor inhibitor, dorsomorphin. Furthermore, knockdown of the central component of the transforming growth factor-β superfamily signaling pathway, Smad4, using small interfering RNA reversed the suppressive effects of BMP15 on Cx43 expression and GJIC activity. The suppressive effects of BMP15 on Cx43 expression were further confirmed in primary human granulosa-lutein cells obtained from infertile patients undergoing an in vitro fertilization procedure. These findings suggest that oocyte-derived BMP15 decreases GJIC activity between human granulosa cells by down-regulating Cx43 expression, most likely via a Smad-dependent signaling pathway.

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References

Feldberg D, Goldman G, Ashkenazi J, Dicker D, Shelef M, Goldman J . The impact of high progesterone levels in the follicular phase of in vitro fertilization (IVF) cycles: a comparative study. J In Vitro Fert Embryo Transf. 1989; 6(1):11-4. DOI: 10.1007/BF01134575. View

Yu P, Hong C, Sachidanandan C, Babitt J, Deng D, Hoyng S . Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nat Chem Biol. 2007; 4(1):33-41. PMC: 2727650. DOI: 10.1038/nchembio.2007.54. View

Petrocelli T, Lye S . Regulation of transcripts encoding the myometrial gap junction protein, connexin-43, by estrogen and progesterone. Endocrinology. 1993; 133(1):284-90. DOI: 10.1210/endo.133.1.8391423. View

Diaz F, Wigglesworth K, Eppig J . Oocytes determine cumulus cell lineage in mouse ovarian follicles. J Cell Sci. 2007; 120(Pt 8):1330-40. DOI: 10.1242/jcs.000968. View

Perkins G, Goodenough D, Sosinsky G . Formation of the gap junction intercellular channel requires a 30 degree rotation for interdigitating two apposing connexons. J Mol Biol. 1998; 277(2):171-7. DOI: 10.1006/jmbi.1997.1580. View

Laitinen M, Vuojolainen K, Jaatinen R, KETOLA I, Aaltonen J, Lehtonen E . A novel growth differentiation factor-9 (GDF-9) related factor is co-expressed with GDF-9 in mouse oocytes during folliculogenesis. Mech Dev. 1998; 78(1-2):135-40. DOI: 10.1016/s0925-4773(98)00161-0. View

Simon A, Goodenough D, Li E, Paul D . Female infertility in mice lacking connexin 37. Nature. 1997; 385(6616):525-9. DOI: 10.1038/385525a0. View

Cook B, Nekola M, NALBANDOV A . Role of the ovum in follicular luteinization. Endocrinology. 1970; 87(2):286-93. View

Loch-Caruso R, Caldwell V, Cimini M, Juberg D . Comparison of assays for gap junctional communication using human embryocarcinoma cells exposed to dieldrin. Fundam Appl Toxicol. 1990; 15(1):63-74. View

10.

Hasegawa J, Yanaihara A, Iwasaki S, Mitsukawa K, Negishi M, Okai T . Reduction of connexin 43 in human cumulus cells yields good embryo competence during ICSI. J Assist Reprod Genet. 2007; 24(10):463-6. PMC: 3455073. DOI: 10.1007/s10815-007-9155-4. View

11.

Mayerhofer A, Garfield R . Immunocytochemical analysis of the expression of gap junction protein connexin 43 in the rat ovary. Mol Reprod Dev. 1995; 41(3):331-8. DOI: 10.1002/mrd.1080410308. View

12.

Derynck R, Zhang Y . Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 2003; 425(6958):577-84. DOI: 10.1038/nature02006. View

13.

Wiesen J, MIDGLEY Jr A . Changes in expression of connexin 43 gap junction messenger ribonucleic acid and protein during ovarian follicular growth. Endocrinology. 1993; 133(2):741-6. DOI: 10.1210/endo.133.2.8393773. View

14.

Melton C, Zaunbrecher G, Yoshizaki G, Patino R, Whisnant S, Rendon A . Expression of connexin 43 mRNA and protein in developing follicles of prepubertal porcine ovaries. Comp Biochem Physiol B Biochem Mol Biol. 2001; 130(1):43-55. DOI: 10.1016/s1096-4959(01)00403-1. View

15.

Cross E, Thomason R, Martinez M, Hopkins C, Hong C, Bader D . Application of small organic molecules reveals cooperative TGFβ and BMP regulation of mesothelial cell behaviors. ACS Chem Biol. 2011; 6(9):952-61. PMC: 3177035. DOI: 10.1021/cb200205z. View

16.

Granot I, Dekel N . The ovarian gap junction protein connexin43: regulation by gonadotropins. Trends Endocrinol Metab. 2002; 13(7):310-3. DOI: 10.1016/s1043-2760(02)00623-9. View

17.

Granot I, Dekel N . Developmental expression and regulation of the gap junction protein and transcript in rat ovaries. Mol Reprod Dev. 1997; 47(3):231-9. DOI: 10.1002/(SICI)1098-2795(199707)47:3<231::AID-MRD1>3.0.CO;2-M. View

18.

Wright C, Becker D, Lin J, Warner A, Hardy K . Stage-specific and differential expression of gap junctions in the mouse ovary: connexin-specific roles in follicular regulation. Reproduction. 2001; 121(1):77-88. DOI: 10.1530/rep.0.1210077. View

19.

Otsuka F, Yao Z, Lee T, Yamamoto S, Erickson G, Shimasaki S . Bone morphogenetic protein-15. Identification of target cells and biological functions. J Biol Chem. 2000; 275(50):39523-8. DOI: 10.1074/jbc.M007428200. View

20.

Hendarto H, Prabowo P, Moeloek F, Soetjipto S . Growth differentiation factor 9 concentration in the follicular fluid of infertile women with endometriosis. Fertil Steril. 2009; 94(2):758-60. DOI: 10.1016/j.fertnstert.2009.10.011. View