Functional expression of the recombinant human FSH receptor.
The functional capacity of the recombinant human FSH (hFSH) receptor was tested on the basis of gonadotrophin stimulation of cyclic AMP (cAMP) production by transient transfections of 293 cells and stable transfections of Chinese hamster ovary (CHO) cells. A CHO cell line expressed with the hFSH receptor cDNA covering the entire amino acid coding region revealed the presence of FSH binding site (Kd 6.2 x 10(-10) M) on the plasma membrane. Treatment of transfected cells with hFSH induced dose-dependent increases in intracellular cAMP production. These results indicate that the hFSH receptor functionally couples with endogenous adenylyl cyclase. Although rat FSH also induced dose-dependent increases in cAMP production, bovine FSH was effective only at high doses and human chorionic gonadotropin did not alter cAMP levels compared with control values. Northern blot analysis with a cRNA probe derived from hFSH receptor cDNA indicated the presence of two common FSH receptor mRNA transcripts (2.4 and 4.1 kb) in RNA prepared from a human ovary and transfected cell lines. Preincubation of CHO cells expressing a functional hFSH receptor (CHO-FSHR) with FSH for 16 h decreased the subsequent cAMP production resulting from a 30-min pulse of FSH stimulation. These results indicate that desensitization of the adenylyl cyclase response to FSH stimulation occurs in CHO-FSHR cells. This cell line therefore provides a tool with which to pursue detailed studies on the molecular basis of FSH-induced desensitization.
Minegishi
T
T
Department of Obstetrics and Gynecology, Gunma University School of Medicine, Japan.
Igarashi
S
S
Nakamura
K
K
Nakamura
M
M
Tano
M
M
Shinozaki
H
H
Miyamoto
K
K
Ibuki
Y
Y
D000818:Animals; D015152:Blotting, Northern; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D000242:Cyclic AMP / Q000096:biosynthesis; D005640:Follicle Stimulating Hormone / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000196:embryology; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis; D011962:Receptors, FSH / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013268:Stimulation, Chemical; D014162:Transfection
D012333:RNA, Messenger; D011962:Receptors, FSH; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP