Corticotropin-releasing hormone gene expression in primary placental cells is modulated by cyclic adenosine 3',5'-monophosphate.
CRH, the principal neuropeptide regulator of pituitary ACTH secretion, is also expressed in placenta. Placental CRH has been linked to the process of human parturition. However, the mechanisms regulating transcription of the CRH gene in placenta remain unclear. cAMP signaling pathways play important roles in regulating the expression of a diverse range of endocrine genes in the placenta. Therefore, we have explored the effect of cAMP on CRH promoter activity in primary cultures of human placental cells. Both forskolin and 8-bromo-cAMP, activators of protein kinase A, can increase CRH promoter activity 5-fold in transiently transfected human primary placental cells, in a manner that parallels the increase in endogenous CRH peptide. Maximal stimulation of CRH promoter activity occurs at 500 micromol/L 8-bromo-cAMP and 10 micromol/L forskolin. Electrophoretic mobility shift assay and mutation analysis combined with transient transfection demonstrate that in placental cells cAMP stimulates CRH gene expression through a cAMP regulatory element in the proximal CRH promoter region and involves a placental nuclear protein interacting specifically with the cAMP regulatory element.
Cheng
Y H
YH
Mothers and Babies Research Center, Endocrine Unit, John Hunter Hospital, Newcastle, New South Wales, Australia.
Nicholson
R C
RC
King
B
B
Chan
E C
EC
Fitter
J T
JT
Smith
R
R
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D003346:Corticotropin-Releasing Hormone / Q000096:biosynthesis* / Q000235:genetics; D000242:Cyclic AMP / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics; D004247:DNA / Q000378:metabolism; D004586:Electrophoresis; D005260:Female; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D016296:Mutagenesis / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism; D010920:Placenta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011247:Pregnancy; D011817:Rabbits; D014162:Transfection / Q000235:genetics
D009687:Nuclear Proteins; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D004247:DNA; D003346:Corticotropin-Releasing Hormone; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases