Forskolin increases neuregulin receptors in human Schwann cells without increasing receptor mRNA.
Forskolin and heregulin synergistically drive human Schwann cell (HSC) proliferation in vitro, but the role of forskolin is not completely understood. To learn how forskolin might affect receptor levels in HSC cultured from adult nerve roots, we first studied expression and localization of HER2 and HER3 in intact roots, using Western blotting and light and electron microscopic immunocytochemistry. We then determined the effect of forskolin and heregulin on receptor expression in HSC cultured from nerve roots using Western blotting and RNase protection assays. HER2 and HER3 were expressed in nonmyelinating Schwann cells in roots and in cultured HSCs before exposure to forskolin. HER2, but not HER3, was also expressed in endoneurial fibroblasts and in cultured nerve root-derived fibroblasts. Treatment with forskolin for 24 h consistently increased HER2 and HER3 protein levels in HSCs but did not alter HER2 and HER3 mRNA levels. In addition, 24-h treatment with heregulin alone decreased HER2 and HER3 protein levels, an effect not previously described. When both heregulin and forskolin were present, HER2 and HER3 protein levels were similar to initial control values. The effect of forskolin on receptor levels was mimicked by dibutyryl-cAMP and receptor levels in both untreated and forskolin treated HSCs were decreased by treatment with the protein kinase A inhibitor H-89. Following pretreatment of HSCs with forskolin, increased receptor levels were correlated with increased rates of thymidine incorporation into HSCs. These results suggest that forskolin/heregulin synergy might derive, at least in part, from post-transcriptional effects leading to increased steady-state receptor levels.
Fregien
Nevis L
NL
Miami Project to Cure Paralysis, University of Miami School of Medicine, Miami, Florida 33136, USA.
White
Linda A
LA
Bunge
Mary Bartlett
MB
Wood
Patrick M
PM
D003994:Bucladesine / Q000494:pharmacology; D049109:Cell Proliferation; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D020890:Neuregulin-1 / Q000378:metabolism / Q000494:pharmacology; D012323:RNA Processing, Post-Transcriptional / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000187:drug effects* / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020893:Receptor, ErbB-3 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012583:Schwann Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013126:Spinal Nerve Roots / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013936:Thymidine / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D003994:Bucladesine; D005576:Colforsin; D017868:Cyclic AMP-Dependent Protein Kinases; D004791:Enzyme Inhibitors; D020890:Neuregulin-1; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D020893:Receptor, ErbB-3; D013936:Thymidine