CHOLINERGIC REGULATION OF VIP GENE-EXPRESSION IN HUMAN NEUROBLASTOMA-CELLS

Citation
B. Kristensen et al., CHOLINERGIC REGULATION OF VIP GENE-EXPRESSION IN HUMAN NEUROBLASTOMA-CELLS, Brain research, 775(1-2), 1997, pp. 99-106
Citations number
28
Language
INGLESE
art.tipo
Article
Journal title
ISSN journal
0006-8993
Volume
775
Issue
1-2
Year of publication
1997
Pages
99 - 106
Database
ISI
SICI code
0006-8993(1997)775:1-2<99:CROVGI>2.0.ZU;2-#
Abstract
The effects of carbamylcholine (CCh) on the gene expression of the neu ropeptide vasoactive intestinal polypeptide (VIP) were studied using t wo human neuroblastoma cell lines, NB-1 and BE(2)M17. CCh caused a fas t increase in VIP mRNA level in both cell Lines which was followed by an increase in VIP immunoreactivity. The time-course of the induction of both mRNA and peptide differed, however, between the two cell lines . No morphological changes of the cells were observed during 6 days of stimulation. The effect was mediated by the muscarinic class of acety lcholine receptors, since it could be totally abolished by atropine. S ince CCh caused an accumulation of inositol-1,4,5-triphosphate, it is likely that muscarinic receptor subtype M1, M3 or M5 is involved. Expe riments with the translational inhibitor, cycloheximide, showed that C Ch mediated a direct effect on the VIP gene expression. By combining g el permeation chromatography with radioimmunoassays using antisera spe cific for the various VIP-precursor products, immunoreactive peaks elu ting as the synthetic peptides were found in both cell lines. In addit ion, earlier eluting peaks which could represent partially processed o r extended VIP forms were found. After CCh induction the concentration of all prepro VIP-derived products increased, and there was a tendenc y towards a shift to more fully processed VIP. The findings give new e vidence for a direct regulation of VIP gene expression in human neuron al cells by cholinergic agents. (C) 1997 Elsevier Science B.V.