Human granulosa contain messenger ribonucleic acids encoding insulin-like growth factor-binding proteins (IGFBPs) and secrete IGFBPs in culture.
OBJECTIVE
To determine if luteinizing human granulosa cells contain messenger ribonucleic acid (mRNAs) encoding insulin-like growth factor-binding protein (IGFBPs) and if cultured granulosa secrete IGFBPs into conditioned medium (CM).


METHODS
Northern analysis, using IGFBP-specific complementary deoxyribonucleic acid probes, was used to detect granulosa-derived IGFBP mRNAs. Western ligand blot analysis of CM was used to detect IGFBPs secreted by granulosa cultures with and without human chorionic gonadotropin (hCG).


METHODS
Granulosa cells were obtained from the In Vitro Fertilization (IVF) Program at Stanford University, a private teaching institution.


METHODS
Patients undergoing IVF for tubal disease.


METHODS
None.


METHODS
Transcripts of IGFBP mRNA and IGFBPs secreted into CM were detected by autoradiography of Northern and Western ligand blots, respectively.


RESULTS
Transcripts of IGFBP-3, IGFBP-2, and IGFBP-1 mRNA were detected in human luteinizing granulosa. Cultured granulosa secreted IGFBPs with molecular weights corresponding to IGFBP-3, IGFBP-2, and IGFBP-1, and the latter two IGFBPs increased with 10 ng/mL hCG. A 24 kd IGFBP was noted, which may be newly characterized IGFBP-4.


CONCLUSIONS
These data show that luteinizing human granulosa cells express mRNAs encoding three IGFBPs, secrete IGFBPs into culture medium, and that production of at least two of the IGFBPs is hCG-dependent, further supporting a role for the IGF system in human folliculogenesis.
Giudice
L C
LC
Department of Gynecology and Obstetrics, Stanford University Medical Center, California 94305-5317.
Milki
A A
AA
Milkowski
D A
DA
el Danasouri
I
I
D015152:Blotting, Northern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D003470:Culture Media; D004247:DNA; D015342:DNA Probes; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D018969:Insulin-Like Growth Factor Binding Proteins; D008970:Molecular Weight; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D013002:Somatomedins / Q000378:metabolism
D002352:Carrier Proteins; D006063:Chorionic Gonadotropin; D003470:Culture Media; D015342:DNA Probes; D018969:Insulin-Like Growth Factor Binding Proteins; D012333:RNA, Messenger; D013002:Somatomedins; D004247:DNA