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B-Type Natriuretic Peptide Receptor Expression and Activity Are Hormonally Regulated in Rat Ovarian Cells¹

Department of Medicine, Division of Experimental Medicine, McGill University (A.N., J.G.); the Laboratory of Cardiovascular Biochemistry, CHUM-Campus Hotel-Dieu, Department of Medicine, University of Montreal (A.N., J.G.)
¹ ; and the Department of Physiology and Obstetrics and Gynecology, Royal Victoria Hospital, McGill University (R.F.)
² , Montréal, Québec, Canada H2W 1T8

Address all correspondence and requests for reprints to: Jolanta Gutkowska, Ph.D., Laboratory of Cardiovascular Biochemistry, Hôtel-Dieu du CHUM, 3850 St. Urbain Street, Pavillon Masson, Montréal, Québec, Canada H2W 1T8. E-mail: jolanta.gutkowska{at}umontreal.ca

Natriuretic peptides form a family of structurally related peptides known to regulate salt and water homeostasis and to cause vasodilation. Synthesis of atrial (ANP), brain (BNP), and C-type (CNP) natriuretic peptides occurs mainly in the heart and brain and has been identified recently in the female reproductive tract. The expression of ANP and CNP as well as their cognate guanylyl cyclase receptors (NPR-A and NPR-B, respectively) have been detected in the rat ovary.

We have shown previously that the expression of the natriuretic peptides and their receptors in the rat ovary appears to be modulated by the estrous cycle. In the present study we have evaluated the expression of the natriuretic peptide system (peptide and receptor) in ovarian cells (granulosa and thecal-interstitial cells) obtained from immature female rats treated with either diethylstilbestrol (DES), an estrogen analog, or equine CG (eCG), a gonadotropin that possesses both LH and FSH activity.

Using a whole cell RRA, we found that CNP binding was increased by 2-fold in granulosa cells taken from animals treated with either DES or eCG. Semiquantitative RT-PCR revealed that granulosa cells from DES- or eCG-treated animals have increased levels of NPR-B messenger RNA (mRNA) transcripts, which was in good agreement with the increased binding. The activity of the receptors was assessed by ligand-dependent stimulation of cGMP release. CNP, but not ANP, stimulated the release of cGMP from granulosa cells obtained from DES-treated, but not from eCG-treated, animals. The relative levels of CNP mRNA in granulosa cells were unaltered by either DES or eCG treatment. In contrast, CNP mRNA levels were increased more than 2-fold, but only in theca-interstitial from the eCG-treated animals.

Our results indicate that CNP and NPR-B are expressed in the ovary, and their expression is responsive to hormonal treatments. Furthermore, expression of these components of the natriuretic peptide system appears to be compartmentalized, with CNP being derived from the extrafollicular compartment and acting, through NPR-B, on the granulosa cells.

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