Biological actions of monoclonal luteinizing hormone/human chorionic gonadotropin receptor antibodies.
Several recent studies have elucidated the structure of the mammalian LH/hCG receptor; as reported in the present work, we have developed a series of monoclonal antibodies (mAbs) against the rat ovarian LH/hCG receptor using highly purified receptor as immunogen and by screening hybridomas with purified LH/hCG receptors. The mAbs were able to specifically immunoprecipitate LH/hCG receptors from solubilized preparations of rat ovarian membranes as well as from partially purified preparations. Western blotting with mAb P1B4 detected a probable receptor dimer and a receptor fragment in rat and porcine ovarian tissue but not in other tissues. This mAb also partially inhibited hCG binding to rat and porcine ovarian tissues. The receptor mAbs were able to inhibit hCG-induced progesterone synthesis in cultured human and porcine granulosa cells without affecting cAMP- and FSH-induced progesterone synthesis. The mAb P1B4 was used to demonstrate that the majority of ovarian receptors are internalized after hCG treatment and that in pseudopregnant rats receptors are present in the rough endoplasmic reticulum and in microvesicles. Bovine corpus luteal cells also contained P1B4 binding sites, as detected by immunohistochemical technique. Taken together, these results suggest that the mAbs are specific for the LH/hCG receptor, mAb P1B4 recognizes an epitope that is highly conserved among mammals, and this epitope is probably in the extracellular domain.
Indrapichate
K
K
Department of Physiology, University of Nebraska Medical Center, Omaha 68198-4575.
Meehan
D
D
Lane
T A
TA
Chu
S Y
SY
Rao
C V
CV
Johnson
D
D
Chen
T T
TT
Wimalasena
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D003338:Corpus Luteum / Q000378:metabolism; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism; D006801:Humans; D008184:Luteal Cells / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D010053:Ovary / Q000378:metabolism; D051381:Rats; D011974:Receptors, LH / Q000276:immunology* / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis; D013552:Swine
D000911:Antibodies, Monoclonal; D011974:Receptors, LH; D013256:Steroids