Similarities and discrepancies in the signaling pathway for nerve growth factor in an insulin producing cell line and a neural crest-derived cell line.
Like neuronal cells, insulin producing cells (beta cells) possess nerve growth factor (NGF) binding sites and express mRNA coding for the low- and high-affinity NGF receptors, p75NGFR and Trk-A respectively. Although the role of NGF on neuronal cells is well documented, its function on beta cells is still unknown. As a first step towards the elucidation of the role of NGF on beta cells, we have characterized both types of NGF receptors on INS-1 cells, a beta cell line derived from a rat insulinoma and studied some early post-receptor events by comparing the signaling pathway of NGF in those cells and in PC12 cells, a well characterized NGF-responsive cell line. By polymerase chain reaction, immunocytochemistry, cross-linking and Western blot analysis, we clearly demonstrated that Trk-A and p75NGFR, the two NGF receptors expressed in INS-1 cells and PC12 cells are similar. Moreover, upon NGF treatment, Trk-A is phosphorylated on tyrosine residues in both cell types in the same dose- and time-dependent manner. These data clearly demonstrate that the first step of NGF signal transduction is similar in PC12 and INS-1 cells. Although early responsive genes like NGFI-A and c-fos are induced in both cell types upon NGF treatment, the induction of c-jun expression is restricted to PC12 cells. Furthermore, the expression of late responsive genes, such as vgf and transin, which are induced by NGF in PC12 cells, are not induced in INS-1 cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Tazi
A
A
INSERM CJF 93-13 Hospital Robert Debré, Paris, France.
Czernichow
P
P
Scharfmann
R
R
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003432:Cross-Linking Reagents; D017781:Genes, Immediate-Early / Q000235:genetics; D007150:Immunohistochemistry; D007328:Insulin / Q000096:biosynthesis*; D007457:Iodine Radioisotopes; D007515:Islets of Langerhans / Q000378:metabolism*; D008969:Molecular Sequence Data; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis* / Q000302:isolation & purification; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014158:Transcription, Genetic
D003432:Cross-Linking Reagents; D007328:Insulin; D007457:Iodine Radioisotopes; D012333:RNA, Messenger; D017475:Receptors, Nerve Growth Factor