Reference - Detail
|Author||Jiang C, Hou X, Wang C, May JV, Butnev VY, Bousfield GR, Davis JS.|
|Title||Hypoglycosylated hFSH Has Greater Bioactivity Than Fully Glycosylated Recombinant hFSH in Human Granulosa Cells.|
|Journal||J Clin Endocrinol Metab|
CONTEXT:Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH.
OBJECTIVE:Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells.
DESIGN AND SETTING:Recombinant human FSH (hFSH) derived from GH3 pituitary cells was purified into fractions containing hypoglycosylated hFSH(21/18) and fully glycosylated hFSH(24). The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center.
INTERVENTIONS:Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours.
MAIN OUTCOME MEASURE(S):The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis.
RESULTS:We observed that hypoglycosylated FSH(21/18) was significantly more effective than fully glycosylated FSH(24) at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH(21/18) was also much more effective than hFSH(24) on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviral-mediated expression of the endogenous inhibitor of PKA, inhibited FSH(21/18)- and FSH(24)-stimulated CREB phosphorylation, and steroidogenesis.
CONCLUSIONS:Hypoglycosylated FSH(21/18) has greater bioactivity than fully glycosylated hFSH(24), suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.
|MeSH||Carbohydrate Sequence Cells, Cultured Cyclic AMP / metabolism Cyclic AMP Response Element-Binding Protein / metabolism Cyclic AMP-Dependent Protein Kinases / metabolism Enzyme Activation / drug effects Female Follicle Stimulating Hormone, Human / metabolism* Follicle Stimulating Hormone, Human / pharmacology* Glycosylation Granulosa Cells / drug effects* Granulosa Cells / metabolism Humans Phosphorylation Protein Isoforms Recombinant Proteins / metabolism* Recombinant Proteins / pharmacology*|
|WOS Category||ENDOCRINOLOGY & METABOLISM|
|Human and Animal Cells||KGN(RCB1154)|