Pituitary adelylate cyclase-activating peptide is an activator of vasoactive intestinal polypeptide gene transcription in human neuroblastoma cells

被引：18

作者：

Georg, B ^{[1
]}

Fahrenkrug, J ^{[1
]}

机构：

[1] Univ Copenhagen, Bispebjerg Hosp, Dept Clin Biochem, DK-2400 Copenhagen NV, Denmark

来源：

MOLECULAR BRAIN RESEARCH | 2000年 / 79卷 / 1-2期

关键词：

CRE site; human neuroblastoma cells; pituitary adenylate cyclase activating peptide; transcription; vasoactive intenstinal polypeptide gene; expression;

D O I：

10.1016/S0169-328X(00)00101-7

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

In many ganglia, the neuropeptide pituitary adenylate cyclase-activating peptide (PACAP) innervates nerve cell bodies containing the homologous neuropoptide vasoactive intestinal polypeptide (VIP). We therefore investigated whether PACAP affected the VIP gene expression and elucidated the molecular mechanisms using the human neuroblastoma cell line NB-1. A concentration dependent induction of the VIP mRNA level was found upon PACAP stimulation. Five nM PACAP mediated transient elevation of the VIP mRNA being evident after 2 h, the maximal 65-fold induction was reached after 6-8 h and hereafter the level decreased rapidly. In cell extracts, the concentration of immunoreactive VIP was elevated four-fold upon PACAP stimulation for 8 h, and it remained elevated during the next 30 h. In conditioned medium, a stable 20-fold VIP increase was seen after 8-24 h. Experiments with the translational inhibitor cycloheximide showed a direct effect of PACAP on the VIP mRNA level, and nuclear run-on assays revealed a three- to four-fold enhancement of the VIP gene transcription rate after PACAP stimulation. The VIP mRNA induction was abolished by transcriptional inhibition with the actinomycin D, and PACAP did not seem to mediate any changes in the VIP mRNA half-life. However, the VIP mRNA level seemed very stable during the transcriptional cessation. Reporter gene constructs were used to evaluate involvement of the VIP CRE site in the PACAP mediated induction of the VIP gene transcription. Mutation of the CRE site did not abolish the induction suggesting it to be of minor if any importance for the induction. In conclusion, the PACAP mediated induction of the VIP gene expression suggests that PACAP released from nerve terminals could influence the function of VIP'ergic neurons in target tissues. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：67 / 76

页数：10

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