title: Protein kinase B is obligatory for follicle-stimulating hormone-induced granulosa cell differentiation.
abstract: Although FSH receptors are linked to the cAMP second messenger system, additional intracellular signaling pathways appear to be required for the induction of aromatase and the LH receptor during granulosa cell differentiation. We employed adenovirus vectors to modulate specific intracellular signaling systems in undifferentiated granulosa cells to identify the signaling pathway(s) that may be involved in the FSH-mediated induction of aromatase and the LH receptor. Expression of either the constitutively activated human LH receptor D578H or the constitutively active human G(s)alpha Q227L resulted in increased cAMP production without increasing aromatase activity or mRNA levels for the LH receptor. To explore the contributions of other pathways, we expressed the constitutively activated forms MAPK kinase (MEK) and protein kinase B (PKB). Neither MEK nor PKB alone increased estrogen or progesterone production by undifferentiated granulosa cells. Stimulation of granulosa cells by FSH in the presence of the constitutively active PKB, but not MEK, led to an amplification of FSH-induced aromatase and LH receptor mRNA levels, whereas a dominant negative PKB vector completely abolished the actions of FSH. The expression of the constitutively active PKB in combination with the constitutively active LH receptor D578H, the constitutively active G(s)alpha Q227L, or 8-bromo-cAMP led to an induction of aromatase as well as LH receptor mRNA comparable to that seen in cells stimulated with FSH alone. These results demonstrate that PKB is an essential component of the FSH-mediated granulosa cell differentiation and that both PKB and G(s)alpha signaling pathways are required.
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA. zeleznik@pitt.edu
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Zeleznik
authors.forename: Deeksha
authors.initials: D
authors.lastname: Saxena
authors.forename: Lynda
authors.initials: L
authors.lastname: Little-Ihrig
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000730:Androstadienes / Q000494:pharmacology; D001141:Aromatase / Q000378:metabolism; D015415:Biomarkers; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004967:Estrogens / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology; D011374:Progesterone / Q000096:biosynthesis; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D011962:Receptors, FSH / Q000378:metabolism; D011974:Receptors, LH / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013449:Sulfonamides; D000077191:Wortmannin
chemical_list: C473217:8-(4-chloro-phenylthio)-2'-O-methyladenosine-3'-5'-cyclic monophosphate; D000730:Androstadienes; D015415:Biomarkers; D004791:Enzyme Inhibitors; D004967:Estrogens; D007546:Isoquinolines; D011518:Proto-Oncogene Proteins; D011962:Receptors, FSH; D011974:Receptors, LH; D013449:Sulfonamides; D011374:Progesterone; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP; D001141:Aromatase; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D019205:GTP-Binding Protein alpha Subunits, Gs; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D000077191:Wortmannin