Functional Dopamine-1 Receptors and DARPP-32 Are Expressed in Human Ovary and Granulosa Luteal Cells in Vitro1
Artur Mayerhofer,
Hugh C. Hemmings, Jr.,
Gretchen L. Snyder,
Paul Greengard,
Sylvia Boddien,
Ulrike Berg and
Cosima Brucker
Anatomisches Institut der Technischen Universität
München (A.M., S.B.), D-80802 München, Germany; Departments
of Anesthesiology and Pharmacology (H.C.H.), Cornell University Medical
College, New York, New York 10021; The Rockefeller University (G.L.S.,
P.G.), New York, New York 10021; and I. Frauenklinik der
Ludwig-Maximilians Universität München (U.B., C.B.),
D-80337 München, Germany
Address all correspondence and requests for reprints to: Artur Mayerhofer, M.D., Professor of Molecular Anatomy, Anatomisches Institut, Technische Universität München, Biedersteiner Strasse 29, D-80802 München, Germany. E-mail:
mayerhofer{at}lrz.tum.de
The catecholamines norepinephrine and dopamine (DA) are present
inthe human ovary; in particular, in follicular fluid. Norepinephrine
activatesovarian - and ß-adrenergic receptors and modulates
ovariansteroidogenesis, but the significance of ovarian DA is unclear.
Weexamined whether a DA receptor of the D1-subtype (D1-R) is present
inhuman ovary and in cultured human granulosa luteal cells (GC).Using
RT-PCR, we cloned complementary DNAs from adult humanovarian and GC
messenger RNAs, which are identical to the humanD1-R sequence. In
ovarian sections, D1-R protein was identified(by immunohistochemistry)
in granulosa cells of large antralfollicles, cells of the corpus
luteum, as well as in culturedGC. An immunoreactive band of
approximately Mr 50,000 was foundin cultured luteinized GC using the
same antiserum in Westernblots. The D1-R in these cells was
functional, because DA, aloneor in the presence of the ß-receptor
antagonist propranolol,caused cellular contraction. The selective D1-R
agonist SKF-38393induced a similar change in cytomorphology and
increased thelevels of media cAMP. SKF-38393 failed, however, to
significantlyaffect basal and hCG-stimulated progesterone
release in vitro,indicating that the activation of the
D1-R was not directlylinked to synthesis of progesterone,
the major steroid of humanGC. Estradiol synthesis likewise was not
affected. Using RT-PCRand immunohistochemistry, we found that GC
express DA- and cAMP-regulatedphosphoprotein of Mr 32,000 (DARPP-32),
a protein typicallyassociated with neurons bearing the D1-R. In
cultured GC, DAand SKF-38393 induced increased
threonine-phosphorylation ofDARPP-32, even in the presence of
propranolol but not in thepresence of D1-R antagonist SCH-23390. Taken
together, the presenceof DA and a functional DA receptor and DARPP-32
indicate thata novel, physiological regulatory pathway involving DA
existsin the human ovary.
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- and ß-adrenergic receptors and modulates
ovarian steroidogenesis, but the significance of ovarian DA is unclear.
We examined whether a DA receptor of the D1-subtype (D1-R) is present
in human ovary and in cultured human granulosa luteal cells (GC). Using
RT-PCR, we cloned complementary DNAs from adult human ovarian and GC
messenger RNAs, which are identical to the human D1-R sequence. In
ovarian sections, D1-R protein was identified (by immunohistochemistry)
in granulosa cells of large antral follicles, cells of the corpus
luteum, as well as in cultured GC. An immunoreactive band of
approximately Mr 50,000 was found in cultured luteinized GC using the
same antiserum in Western blots. The D1-R in these cells was
functional, because DA, alone or in the presence of the ß-receptor
antagonist propranolol, caused cellular contraction. The selective D1-R
agonist SKF-38393 induced a similar change in cytomorphology and
increased the levels of media cAMP. SKF-38393 failed, however, to
significantly affect basal and hCG-stimulated progesterone
release in vitro, indicating that the activation of the
D1-R was not directly linked to synthesis of progesterone,
the major steroid of human GC. Estradiol synthesis likewise was not
affected. Using RT-PCR and immunohistochemistry, we found that GC
express DA- and cAMP-regulated phosphoprotein of Mr 32,000 (DARPP-32),
a protein typically associated with neurons bearing the D1-R. In
cultured GC, DA and SKF-38393 induced increased
threonine-phosphorylation of DARPP-32, even in the presence of
propranolol but not in the presence of D1-R antagonist SCH-23390. Taken
together, the presence of DA and a functional DA receptor and DARPP-32
indicate that a novel, physiological regulatory pathway involving DA
exists in the human ovary. 
