Granulosa cell luteinizing hormone receptor expression is modulated by ganglioside-specific ligands.
The ganglioside GM1 (Gal beta 1-->3GalNAc beta 1-->4[NeuAc alpha 2-->3] Gal beta 1-->4Glc beta 1-->1Cer) was synthesized during granulosa cell development in vitro, and the effect of the interaction between cell-surface GM1 and its ligands on the luteinizing hormone (LH) receptor expression was investigated. GM1 synthesis, demonstrated by metabolic labeling of glycosphingolipids with [3H]galactose and binding studies using the 125I-B-subunit of cholera toxin, a specific ligand for GM1, was increased in follicle-stimulating hormone (FSH)-treated granulosa cells. When granulosa cells were cultured for 72 h in a medium containing the B-subunit of cholera toxin, FSH-induced LH-receptor contents determined by measuring the binding of 125I-deglycosylated human chorionic gonadotropin to intact cells, was augmented. The stimulatory effect of the B-subunit was dependent on the FSH concentration and culture duration. The augmentation was observed after culture for 48 h, and marked increases were evident after 72 h, which coincided with an increase of the 125I-B-subunit binding capacity. Scatchard analysis of the LH-receptor binding indicated that treatment with the B-subunit increased the number of LH-binding sites (6580 sites/cell after treatment with 20 ng/ml FSH; 11,290 sites/cell after FSH plus 100 ng/ml B-subunit), but did not alter the binding affinity. A specific antibody against GM1 mimicked the stimulatory effect of the B-subunit. The augmentation was not accompanied by granulosa cell proliferation. These findings suggest that binding of exogenous or possible endogenous ligands to cell-surface GM1 produces signals and modulates the cellular behavior during granulosa cell development.
Hattori
M
M
Institute of Endocrinology, Gunma University, Japan.
Kanzaki
M
M
Kojima
I
I
Horiuchi
R
R
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000378:metabolism / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004054:Diethylstilbestrol / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006028:Glycosphingolipids / Q000378:metabolism; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008563:Membrane Lipids / Q000096:biosynthesis / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D011974:Receptors, LH / Q000096:biosynthesis*
D000906:Antibodies; D006028:Glycosphingolipids; D008563:Membrane Lipids; D011974:Receptors, LH; D005677:G(M1) Ganglioside; D004054:Diethylstilbestrol; D005640:Follicle Stimulating Hormone; D004247:DNA; D002772:Cholera Toxin; D000242:Cyclic AMP