Characterization of insulin-like growth factor binding to human granulosa cells obtained during in vitro fertilization.
Insulin and IGF-I affect in vitro ovarian stromal and follicular cell function in several species. We previously characterized insulin receptors on human granulosa cells obtained from in vitro fertilization procedures but were unable to demonstrate specific binding of IGF-I. Following modification of the assay conditions, we now report specific, high affinity IGF-1 binding sites on human granulosa cells. Substitution of equimolar concentrations of sucrose for sodium chloride in the buffer solution increased binding of IGF but not insulin in equilibrium assays. Maximal specific IGF-I binding was 2.69 +/- 0.30%/10(5) cells (SEM, n = 9) with half-maximal inhibition of binding at 2 ng/ml IGF-I. Unlabeled insulin recognized the type I IGF receptor with low affinity. An IGF-I receptor monoclonal antibody (alpha IR-3) inhibited 125I-IGF-I but not 125I-insulin binding. Affinity crosslinking followed by SDS/PAGE under reducing conditions revealed IGF-I binding at a molecular weight compatible with the alpha subunit of the type I IGF receptor and with a pattern of inhibition by various ligands that paralleled the equilibrium binding assays. IGF-I receptors are present on freshly isolated human ovarian granulosa cells obtained following pharmacologic stimulation with gonadotrophin according to the protocols of in vitro fertilization. The biologic function of these receptors currently is being investigated.
Gates
G S
GS
Charles A. Dana Research Institute, Boston, Mass.
Bayer
S
S
Seibel
M
M
Poretsky
L
L
Flier
J S
JS
Moses
A C
AC
D000345:Affinity Labels; D001665:Binding Sites; D002021:Buffers; D005260:Female; D005307:Fertilization in Vitro; D006107:Granulosa Cells / Q000032:analysis*; D006801:Humans; D010053:Ovary / Q000032:analysis; D011247:Pregnancy; D013002:Somatomedins / Q000302:isolation & purification*
D000345:Affinity Labels; D002021:Buffers; D013002:Somatomedins