Gonadotropin regulation of testosterone production by primary cultured theca and granulosa cells of Atlantic croaker: II. Involvement of a mitogen-activated protein kinase pathway.
Previous investigations in Atlantic croaker ovaries and primary co-cultured theca and granulosa cells have identified multiple signal transduction pathways involved in the control of gonadotropin-induced steroidogenesis, including adenylyl cyclase- and calcium-dependent signaling pathways. In the present study, evidence was obtained for an involvement of a third signal transduction pathway, a mitogen-activated protein kinase (MAP kinase) signaling cascade, in the regulation of gonadal steroidogenesis in this lower vertebrate teleost model. Gonadotropin-stimulated testosterone synthesis was markedly attenuated by two antagonists of mitogen-activated protein kinase kinases 1/2 (MEK1/2, also known as Map2k1/Map2k2). Moreover, treatment with gonadotropin-induced MEK1/2-dependent phosphorylation of extracellular signal-regulated protein kinases 1/2 (ERK1/2, also known as Mapk3/Mapk1) in a concentration- and time-dependent manner in co-cultured croaker theca and granulosa cells. Active MEK1/2 was required for a complete steroidogenic response to activators of the adenylyl cyclase pathway, including forskolin and dbcAMP, suggesting that the target(s) of MAP kinase signaling are distal to cAMP generation and activation of cAMP-dependent protein kinase (PKA). Interestingly, dbcAMP caused a similar increase of ERK1/2 phosphorylation as was observed with gonadotropin treatment, although an inhibitor of PKA did not attenuate this response. Finally, there was no evidence of cross-talk between calcium-dependent signaling pathways and this MAP kinase cascade. While drugs that block calcium-dependent signal transduction, including inhibitors of voltage-sensitive calcium channels, calmodulin, and calcium/calmodulin-dependent kinases, significantly reduced gonadotropin-induced testosterone accumulation, these drugs had no apparent effect on hCG-induced ERK1/2 phosphorylation.
Department of Marine Science, University of Texas at Austin, 750 Channel View Drive, Port Aransas, TX 78373, USA. Abby.Benninghoff@science.oregonstate.edu
Abby D
AD
Benninghoff
Peter
P
Thomas
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D018920:Coculture Techniques / Q000379:methods; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis; D017868:Cyclic AMP-Dependent Protein Kinases / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology*; D005260:Female; D005419:Flavonoids / Q000494:pharmacology; D006062:Gonadotropins / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D048370:MAP Kinase Kinase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D008954:Models, Biological; D006080:Ovarian Follicle / Q000254:growth & development; D010473:Perciformes / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013739:Testosterone / Q000096:biosynthesis*; D013799:Theca Cells / Q000187:drug effects / Q000378:metabolism*
D005419:Flavonoids; D006062:Gonadotropins; D005576:Colforsin; D013739:Testosterone; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D048052:Mitogen-Activated Protein Kinase 3; D048369:MAP Kinase Kinase 1; D048370:MAP Kinase Kinase 2; D000262:Adenylyl Cyclases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D002118:Calcium