Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology

Production and binding of endothelin-2 (EDN2) in the rat ovary: endothelin receptor subtype A (EDNRA)-mediated contraction

Phillip J. Bridges A , Misung Jo B , Linah Al Alem A , Giyoun Na A , Wen Su C , Ming C. Gong C , Myoungkun Jeoung A and CheMyong Ko A D E

A Division of Clinical and Reproductive Sciences, University of Kentucky, Lexington, KY 40536, USA.

B Department of Obstetrics and Gynecology, University of Kentucky, Lexington, KY 40536, USA.

C Department of Physiology, University of Kentucky, Lexington, KY 40536, USA.

D Department of Biology, University of Kentucky, Lexington, KY 40536, USA.

E Corresponding author. Email:

Reproduction, Fertility and Development 22(5) 780-787
Submitted: 18 August 2009  Accepted: 11 November 2009   Published: 7 April 2010


Endothelin-2 (EDN2)-mediated contraction has been proposed as a final mechanical signal facilitating ovulation. The objectives herein were to determine (1) whether ovarian endothelins were increased before ovulation; (2) whether a specific endothelin-converting enzyme (ECE) was mediating their production; (3) which receptor was facilitating ovarian contraction; and (4) whether receptor-specific antagonism affected ovulation. Follicular development was induced in immature rats with 10 IU pregnant mare serum gonadotrophin (PMSG) and the ovulatory cascade was initiated 48 h later with 10 IU human chorionic gonadotrophin (hCG). In Experiment 1, an immunoassay revealed that the ovarian concentration of endothelin peptide was increased 7-fold 12 h after hCG when compared with 48 h after PMSG (P < 0.05). In Experiment 2, real-time PCR indicated that mRNA for Ece1, but not Ece2, was increased in granulosa cells collected 12 h after hCG when compared with those collected before the ovulatory stimulus (P < 0.05). In Experiment 3, isometric tension analysis revealed that the contractile effect of EDN2 was mediated by endothelin receptor A (EDNRA), not B (EDNRB). In Experiment 4, no effect was observed on the rate of ovulation when rats were treated with an antagonist specific to EDNRA (BQ123) or EDNRB (BQ788), or when mice were treated with BQ123, BQ788 or BQ123 + BQ788. In conclusion, endothelin peptide is produced before ovulation and the contractile action of EDN2 within the ovary is facilitated via EDNRA. In addition, findings of this study indicate synergistic interactions among contractile factors affect ovulatory outcome, while the role of EDNRB alone in the process of ovulation requires further investigation.

Additional keywords: constriction, ovulation, rupture.


Acosta T. J.Berisha B.Ozawa T.Sato K.Schams D.Miyamoto A.1999Evidence for a local endothelin–angiotensin–atrial natriuretic peptide system in bovine mature follicles in vitro: effects on steroid hormones and prostaglandin secretion.Biol. Reprod.6114191425doi:10.1095/BIOLREPROD61.6.1419Pubmed Abstract

Al-Alem L.Bridges P. J.Su W.Gong M. C.Iglarz M.Ko C.2007Endothelin-2 induces oviductal contraction via endothelin receptor subtype A in rats.J. Endocrinol.193383391doi:10.1677/JOE-07-0089Pubmed Abstract

Alexander M. J.Mahoney P. D.Ferris C. F.Carraway R. E.Leeman S. E.1989Evidence that neurotensin participates in the central regulation of the preovulatory surge of luteinizing hormone in the rat.Endocrinology124783788doi:10.1210/ENDO-124-2-783Pubmed Abstract

Chester A. H.Azam R.Felkin L. E.George R.Brand N.2007Correlation between vascular responsivensss and expression of novel transcripts of the ETA-receptor in human vascular tissue.Vascul. Pharmacol.46181doi:10.1016/J.VPH.2006.09.007Pubmed Abstract

Chun S. Y.Popliker M.Reich R.Tsafriri A.1992Localization of preovulatory expression of plasminogen activator inhibitor type-1 and tissue inhibitor of metalloproteinase type-1 mRNAs in the rat ovary.Biol. Reprod.47245253doi:10.1095/BIOLREPROD47.2.245Pubmed Abstract

Continho E. M.Maia H. S.1971The contractile response of the human uterus, fallopian tubes, and ovary to prostaglandins in vivo.Fertil. Steril.22539543Pubmed Abstract

Coutinho E. M.1974Ovarian contractility and ovulation.Res. Reprod.634Pubmed Abstract

Curry T. E.JrSmith M. F.2006Impact of extracellular matrix remodeling on ovulation and the folliculo–luteal transition.Semin. Reprod. Med.24228241doi:10.1055/S-2006-948552Pubmed Abstract

Debeljuk L.2006Tachykinins and ovarian function in mammals.Peptides27736doi:10.1016/J.PEPTIDES.2005.08.002Pubmed Abstract

Espey L. L.Lipner H.1963Measurements of intrafollicular pressures in the rabbit ovary.Am. J. Physiol.20510671072Pubmed Abstract

Flores J. A.2000Gene expression of endothelin-1 in the porcine ovary: follicular development.Biol. Reprod.6313771382Pubmed Abstract

Gómez R.Simón C.Remohí J.Pellicer A.2003Administration of moderate and high doses of gonadotropins to female rats increases ovarian vascular endothelial growth factor (VEGF) and VEGF Receptor-2 expression that is associated to vascular hyperpermeability.Biol. Reprod.6821642171doi:10.1095/BIOLREPROD.102.010801Pubmed Abstract

Guo Z.Su W.Ma Z.Smith G. M.Gong M. C.2003Ca2+-independent phospholipase A2 is required for agonist-induced Ca2+ sensitization of contraction in vascular smooth muscle.J. Biol. Chem.27818561863doi:10.1074/JBC.M211075200Pubmed Abstract

Hasan S.Hosseini G.Princivalle M.Dong J.-C.Birsan D.Cagide Agostini A. I.2002Coordinate expression of anticoagulant heparan sulfate proteoglycans and serine protease inhibitors in the rat ovary: a potent system of proteolysis control.Biol. Reprod.66144158doi:10.1095/BIOLREPROD66.1.144Pubmed Abstract

Hellberg P.Larson L.Olofsson J.Hedin L.Brannstrom M.1991Stimulatory effects of bradykinin on the ovulatory process in the in vitro-perfused rat ovary.Biol. Reprod.44269274doi:10.1095/BIOLREPROD44.2.269Pubmed Abstract

Hoang M. V.Turner A. J.1997Novel activity of endothelin-converting enzyme: hydrolysis of bradykinin.Biochem. J.327Pt 12326Pubmed Abstract

Johnson G. D.Stevenson T.Ahn K.1999Hydrolysis of peptide hormones by endothelin-converting enzyme-1. A comparison with neprilysin.J. Biol. Chem.27440534058doi:10.1074/JBC.274.7.4053Pubmed Abstract

Kitajima Y.Endo T.Manase K.Nishikawa A.Shibuya M.Kudo R.2004Gonadotropin-releasing hormone agonist administration reduced vascular endothelial growth factor (VEGF), VEGF receptors, and vascular permeability of the ovaries of hyperstimulated rats.Fertil. Steril.81842doi:10.1016/J.FERTNSTERT.2003.11.012Pubmed Abstract

Ko C.Gieske M. C.Al-Alem L.Hahn Y.Su W.Gong M. C.Iglarz M.Koo Y.2006Endothelin-2 in ovarian follicle rupture.Endocrinology14717701779doi:10.1210/EN.2005-1228Pubmed Abstract

Liu Y. X.Peng X. R.Chen Y. J.Carrico W.Ny T.1997Prolactin delays gonadotrophin-induced ovulation and down-regulates expression of plasminogen-activator system in ovary.Hum. Reprod.1227482755doi:10.1093/HUMREP/12.12.2748Pubmed Abstract

Livak K. J.Schmittgen T. D.2001Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method.Methods25402408doi:10.1006/METH.2001.1262Pubmed Abstract

Masaki T.Ninomiya H.Sakamoto A.Okamoto Y.1999Structural basis of the function of endothelin receptor.Mol. Cell. Biochem.190153156doi:10.1023/A:1006986914326Pubmed Abstract

Matousek M.Carati C.Gannon B.Brannstrom M.2001Novel method for intrafollicular pressure measurements in the rat ovary: increased intrafollicular pressure after hCG stimulation.Reproduction121307314doi:10.1530/REP.0.1210307Pubmed Abstract

Miyabayashi K.Shimizu T.Kawauchi C.Sasada H.Sato E.2005Changes of mRNA expression of vascular endothelial growth factor, angiopoietins and their receptors during the periovulatory period in eCG/hCG-treated immature female rats.J. Exp. Zool. A Comp. Exp. Biol.303A590597doi:10.1002/JEZ.A.188

O’Shea J. D.Phillips R. E.1974Contractility in vitro of ovarian follicles from sheep, and the effect of drugs.Biol. Reprod.10370379doi:10.1095/BIOLREPROD10.3.370Pubmed Abstract

Palanisamy G. S.Cheon Y.-P.Kim J.Kannan A.Li Q.Sato M.Mantena S. R.Sitruk-Ware R. L.Bagchi M. K.Bagchi I. C.2006A novel pathway involving progesterone receptor, endothelin-2, and endothelin receptor B controls ovulation in mice.Mol. Endocrinol.2027842795doi:10.1210/ME.2006-0093Pubmed Abstract

Palti Z.Freund M.1972Spontaneous contractions of the human ovary in vitro.J. Reprod. Fertil.28113115doi:10.1530/JRF.0.0280113Pubmed Abstract

Priddy A. R.Killick S. R.1993Eicosanoids and ovulation.Prostaglandins Leukot. Essent. Fatty Acids49827831doi:10.1016/0952-3278(93)90204-APubmed Abstract

Rondell P.1964Follicular pressure and distensibility in ovulation.Am. J. Physiol.207590594Pubmed Abstract

Schmidt G.Kannisto P.Owman C.Walles B.1987Characterization of histamine receptors mediating contraction and relaxation of the ovarian follicle wall.Int. J. Fertil.32399406Pubmed Abstract

Schroeder P. C.Talbot P.1982Intrafollicular pressure decreases in hamster preovulatory follicles during smooth muscle cell contraction in vitro.J. Exp. Zool.224417426doi:10.1002/JEZ.1402240315Pubmed Abstract

Smith C.Perks A. M.1984The effects of bradykinin on the contractile activity of the isolated rat ovary.Acta Endocrinol. (Copenh.)106387392Pubmed Abstract

Talbot P.Chacon R. S.1982In vitro ovulation of hamster oocytes depends on contraction of follicular smooth muscle cells.J. Exp. Zool.224409415doi:10.1002/JEZ.1402240314Pubmed Abstract

Tedeschi C.Hazum E.Kokia E.Ricciarelli E.Adashi E. Y.Payne D. W.1992Endothelin-1 as a luteinization inhibitor: inhibition of rat granulosa cell progesterone accumulation via selective modulation of key steroidogenic steps affecting both progesterone formation and degradation.Endocrinology13124762478doi:10.1210/EN.131.5.2476Pubmed Abstract

Tirapelli C. R.Casolari D. A.Montezano A. C.Yogi A.Tostes R. C.Legros E.D’Orleans-Juste P.Lanchote V. L.Uyemura S. Oliveira A. M.2006Ethanol consumption enhances endothelin-1-induced contraction in the isolated rat carotid.J. Pharmacol. Exp. Ther.318819827doi:10.1124/JPET.106.103010Pubmed Abstract

Turner A. J.Murphy L. J.1996Molecular pharmacology of endothelin-converting enzymes.Biochem. Pharmacol.5191doi:10.1016/0006-2952(95)02036-5Pubmed Abstract

Virutamasen P.Smitasiri Y.Fuchs A. R.1976Intraovarian pressure changes during ovulation in rabbits.Fertil. Steril.27188196Pubmed Abstract

Yoshioka S.Fujiwara H.Yamada S.Tatsumi K.Nakayama T.Higuchi T.Inoue T.Maeda M.Fujii S.1998Endothelin-converting enzyme-1 is expressed on human ovarian follicles and corpora lutea of menstrual cycle and early pregnancy.J. Clin. Endocrinol. Metab.8339433950doi:10.1210/JC.83.11.3943Pubmed Abstract

Full Text PDF (272.2 KB) Export Citation