We have previously shown the presence of immunoreactive angiotensin-(1—7) [Ang-(1—7)] in rat ovary homogenate and its stimulatory effect on estradiol and progesterone production in vitro. In the current study, we investigated the presence and cellular distribution of Ang-(1—7) and the Mas receptor, the expression of Mas and angiotensin-converting enzyme 2 (ACE2) messenger RNA (mRNA), and the enzymatic activity in the rat ovary following gonadotropin stimulation in vivo. Immature female Wistar rats (25 days old) were injected subcutaneously (SC) with equine chorionic gonadotropin (eCG, 20 IU in 0.2 mL) or vehicle 48 hours before euthanasia. Tissue distributions of Ang-(1—7), Mas receptor, and ACE2 were evaluated by immunohistochemistry, along with angiotensin II (Ang II) localization, while the mRNA expression levels of Mas receptor and ACE2 were evaluated by real-time polymerase chain reaction (PCR). In addition, we determined the activity of neutral endopeptidase (NEP), prolyl endopeptidase (PEP), and ACE by fluorometric assays. After eCG treatment, we found strong immunoreactivity for Ang-(1—7) and Mas primarily in the theca-interstitial cells, while Ang II appeared in the granulosa but not in the thecal layer. Equine chorionic gonadotropin treatment increased Mas and ACE2 mRNA expression compared with control animals (3.3- and 2.1-fold increase, respectively; P < .05). Angiotensin-converting enzyme and NEP activities were lower, while PEP activity was higher in the eCG-treated rats (P < .05). These data show gonadotropin-induced changes in the ovarian expression of Ang-(1—7), Mas receptor, and ACE2. These findings suggest that the renin-angiotensin system (RAS) branch formed by ACE2/Ang-(1—7)/Mas, fully expressed in the rat ovary and regulated by gonadotropic hormones, could play a role in the ovarian physiology.
Driancourt MA, Gougeon A., Royère D., Thibault C.Ovarian function. In: Thibault C, Levasseur MC, eds. Reproduction in Mammals and Man. Paris: Ellipses; 1993:281-305.
3.
Yoshimura Y.The ovarian renin-angiotensin system in reproductive physiology. Front Neuroendocrinol. 1997;18(3):247-291.
4.
Pellicer A., Palumbo A., DeCherney AH, Naftolin F.Blockade of ovulation by an angiotensin antagonist. Science . 1988; 240(4859):1660-1661.
5.
Yoshimura Y. , Karube M., Aoki H., et al. Angiotensin II induces ovulation and oocyte maturation in rabbit ovaries via the AT2 receptor subtype. Endocrinology. 1996;137(4):1204-1211.
6.
Kuji N., Sueoka K., Miyasaki T., Tanaka M., Oda T., Kobayashi T. Involvement of Angiotensin II in the process of gonadotropin-induced ovulation in rabbits. Biol Reprod. 1996;55(5):984-991.
7.
Daud AI, Bumpus FM, Husain A.Characterization of angiotensin I converting enzyme (ACE)-containing follicles in the rat ovary during the estrous cycle and effects of ACE inhibitor on ovulation. Endocrinology. 1990;126(6):2927-2935.
8.
Yoshimura Y. , Koyama N., Karube M., et al. Gonadotropin stimulates ovarian renin-angiotensin system in the rabbit. J Clin Invest. 1994;93(1):180-187.
9.
Bumpus FM, Pucell AG, Daud AI, Husain A.Angiotensin II, an intraovarian regulatory peptide. Am J Med Sci.1988;295(4): 406-408.
10.
Yoshimura Y. , Karube M., Oda T., et al. Locally produced Angiotensin II induces ovulation by stimulating prostaglandin production in in vitro perfused rabbit ovaries. Endocrinology. 1993;133(4):1609-1616.
11.
Magoffin DALH stimulation of ovarian theca-interstitial cell differentiation: sensitizing effect of angiotensin II. In: Gibori G, ed. Signalling Mechanisms and Gene Expression in the Ovary. New York: Springer-Verlag;1991:417-422.
12.
Fèral C., Gall SL, Leymarie P. Angiotensin II modulates steroidogenesis in granulosa and theca in the rabbit ovary: its possible involvement in atresia . Eur J Endocrinol. 1995;133(4): 747-753.
13.
Aguilera G. , Millan MA, Harwood JPAngiotensin II receptors in the gonads. Am J Hypertens. 1989;2(5 pt 1):395-402.
14.
Peterson CM , Morioka N., Zhu C., Ryan JW, LeMarie WJAngiotensin-converting enzyme inhibitors have no effect on ovulation and ovarian steroidogenesis in the perfused rat ovary . Reprod Toxicol. 1993;7(2):131-135.
15.
Santos RA, Ferreira AJ, Simões e Silva ACRecent advances in the angiotensin-converting enzyme 2-angiotensin(1-7)-Mas axis. Exp Physiol. 2008;93(5):519-527.
16.
Santos RAS , Brosnihan KB, Jacobsen DW, DiCorleto PE, Ferrario CMProduction of angiotensin-(1-7) by human vascular endothelium. Hypertension. 1992 ;19(2 suppl):1156-1161.
17.
Welches WR, Santos RA, Chappell MC, Brosnihan KB, Greene LJ, Ferrario CMEvidence that prolyl endopeptidase participates in the processing of brain angiotensin. J Hypertens. 1991;9(7):631-638.
18.
Neves LAA , Almeida AP, Khosla MC, Santos RASMetabolism of angiotensin I in isolated rat hearts. Effect of angiotensin-converting enzyme inhibitors. Biochem Pharmacol. 1995 ;50(9):1451-1459.
19.
Donoghue M. , Hsieh F., Baronas E., et al. A novel angiotensin-converting enzyme-related carboxypeptidase (ACE2) converts angiotensin I to angiotensin 1-9. Circ Res . 2000;87 (5):E1-E9.
20.
Costa APR , Fagundes-Moura CR, Pereira VM, et al. Angiotensin-(1-7): a novel peptide in the ovary. Endocrinology. 2003;144(5):1942-1948.
21.
Jackson TR, Blair LA, Marshall J., Goedert M., Hanley MRThe Mas oncogene encodes an angiotensin receptor. Nature. 1988;335(6189):437-440.
22.
Metzger R., Bader M., Ludwig T., Berberich C., Bunnemann B., Ganten D.Expression of the mouse and rat Mas proto-oncogene in the brain and peripheral tissues. FEBS Lett. 1995;357(1):27-32.
23.
Santos RA, Simões e Silva AC, Maric C., et al. Angiotensin-(1-7) is a endogenous ligand for the G protein coupled receptor Mas. Proc Natl Acad Sci U S A.2003 ;100(14):8258-8263.
24.
Pereira VM, Costa APR, Rosa-e-Silva AA, Vieira MA, Reis AMRegulation of steroidogenesis by atrial natriuretic peptide (ANP) in the rat testis: differential involvement of GC-A and C receptors. Peptides. 2008;29(11):2024-2032.
25.
Reis FM, Ribeiro MF, Maia AL, Spritzer PMRegulation of human endometrial transforming growth factor beta 1 and beta 3 isoforms through menstrual cycle and medroxyprogesterone acetate treatment . Histol Histopathol. 2002;17(3):739-745.
26.
Vaz-Silva J. , Carneiro MM, Ferreira MC, et al. The vasoactive peptide angiotensin-(1-7), its receptor Mas and the angiotensin-converting-enzyme type 2 are expressed in the human endometrium. Reprod Sci.2009 ;16(3):247-256.
27.
Simões e Silva AC, Diniz JS, Regueira-Filho A., Santos RAThe renin angiotensin system in childhood hypertension: selective increase of angiotensin-(1-7) in essential hypertension. J Pediatr.2004 ;145(1):93-98.
28.
Becker LK, Etelvino GM, Walther T., Santos RA, Campagnole-Santos MJImmunofluorescence localization of the receptor Mas in cardiovascular-related areas of the rat brain. Am J Physiol Heart Circ Physiol. 2007;293(3):H1416-H1424.
29.
Lowry OH, Rosebrough NJ, Farr AL, Randal LJProtein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-275.
30.
Santos RAS , Krieger EM, Greene LJAn improved fluorometric assay of rat serum and plasma converting enzyme. Hypertension. 1985;7(2):244-252.
31.
Stanziola L. , Greene LJ, Santos RAEffect of chronic angiotensin converting enzyme inhibition on angiotensin I and bradikinin metabolism in rats. Am J Hypertens.1999;12(10 pt 1):1021-1029.
32.
Medeiros MAS , França MSF, Boileau G., Juliano L., Carvalho KMSpecific fluorogenic substrates for neprilysin (neutral endopeptidase, EC 3.4.24.11) which are highly resistant to serine- and metalloproteases. Braz J Med Biol Res. 1997;30(10):1157-1162.
33.
Carithers JR , Green JAUltrastructure of rat ovarian interstitial cells. I. Normal structure and regressive changes following hypophysectomy . J Ultrastr Res. 1972;39(3): 239-250.
34.
Erickson GF , Magoffin DA, Dyer CA, Hofeditz C.The ovarian androgen producing cells: a review of structure/function relationships. Endocr Rev. 1985;6(3):371-399.
35.
Ohta N., Takahashi T., Mori T., et al. Hormonal modulation of prolyl endopeptidase and dipeptidyl peptidase IV activities in the mouse uterus and ovary. Acta Endocrinol . 1992;127(3): 262-266.
36.
Pinto FM, Armesto CP, Magraner J., Trujillo M., Martin JD, Cadenas MLTachikinin receptor and neutral endopeptidase gene expression in the rat uterus: characterization and regulation in response to ovarian steroid treatment. Endocrinology. 1999;140(6):2526-2532.
37.
Harata T., Ando H., Iwase A., Nagasaka T., Mizutani S., Kikkawa F. Localization of angiotensin II, the AT1 receptor, angiotensin-converting enzyme, aminopeptidase A, adipocytederived leucine aminopeptidase, and vascular endothelial growth factor in the human ovary throughout the menstrual cycle . Fertil Steril. 2006;86(2):433-439.
38.
Shuttleworth G., Hunter MG, Robinson G., Pipkin FBImmunocytochemical localization of angiotensin II receptor subtypes 1 and 2 in the porcine fetal, prepubertal and postpubertal ovary. J Anat. 2002;201(3):267-274.
39.
Pountain SJ , Pipkin FB, Hunter MGThe ontogeny of components of the renin-angiotensin system in the porcine fetal ovary. Anim Reprod Sci. in press. doi: 10.1016/ j.anireprosci.2009.03.006.
40.
Hayashi M., McGee EA, Min G., et al. Recombinant growth differentiation factor-9 (GDF-9) enhances growth and differentiation of cultured early ovarian follicles. Endocrinology. 1999;140(3):1236-1244.
41.
Solovyeva EV , Hayashi M., Margi K., et al. Growth differentiation factor-9 stimulates rat theca-interstitial cell androgen biosynthesis. Biol Reprod . 2000;63(4):1214-1218.
42.
Pepperell JR , Nemeth G., Yamada Y., Naftolin F., Merino M.Localized accumulation of angiotensin II and production of angiotensin-(1-7) in rat luteal cells and effects on steroidogenesis . Am J Physiol Endocrinol Metab.2006;291(2):E221-E233.
43.
Palumbo A., Jones C., Lightman A., Carcangiu ML, DeCherney AH, Naftolin F.Immunohistochemical localization of renin and angiotensin II in human ovaries . Am J Obstet Gynecol. 1989;160(1):8-14.
44.
Howard RB, Pucell AG, Bumpus FM, Husain A.Rat ovarian renin: characterization and changes during the estrous cycle. Endocrinology. 1988;123(5):2331-2340.
45.
Thomas WG, Sernia C.The immunocytochemical localization of angiotensinogen in the rat ovary. Cell Tissue Res. 1990;261(2):367-373.
46.
Speth RC, Husain A.Distribution of angiotensin-converting enzyme and angiotensin II-receptor binding sites in the rat ovary. Biol Reprod. 1988;38(3):695-702.
