Publications (4)4.06 Total impact
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Article: Synergistic Induction of Pituitary Adenylate Cyclase‐Activating Polypeptide (PACAP) Gene Expression by Nerve Growth Factor and PACAP in PC12 Cells
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ABSTRACT: Pituitary adenylate cyclase-activating polypeptide (PACAP) gene expression was analyzed in PC12 cells. PC12 cells transfected with a PACAP promoter-luciferase reporter construct were utilized to investigate the effects of PACAP, either alone or in combination with nerve growth factor (NGF), on PACAP transcriptional response. PACAP induced transcription from the PACAP promoter through PACAP type I receptor (PAC1 receptor). PACAP gene transcription was also induced by NGF. Simultaneous treatment with PACAP and NGF resulted in a synergistic transcriptional response that was more than three times the predicted response, based on a simple additive effect of both agents. This synergism in transcriptional response paralleled the PACAP mRNA levels, as determined by RT-PCR and northern blotting. The level of PACAP mRNA peaked 3 h after stimulation and gradually returned to basal levels by 48 h. PC12 cells are known to express predominantly the hop isoform of the PAC1 receptor, which positively couples to both adenylate cyclase and phospholipase C. To determine the role of the cyclic AMP and protein kinase C pathways in PACAP gene expression, the effects of forskolin and phorbol 12-myristate 13-acetate (PMA) were then examined. PMA did not alter PACAP mRNA levels but enhanced forskolin-induced PACAP mRNA expression. Down-regulation of protein kinase C blocked the ability of PACAP to stimulate PACAP mRNA expression. The mitogen-activated protein kinase extracellular signal-regulated kinase (ERK) kinase 1/2 (MEK1/2) inhibitor PD98059 also blocked the PACAP mRNA expression induced by either PACAP or NGF but not that induced by a combination of PACAP and NGF. These results suggest that PACAP stimulates the PACAP gene expression in PC12 cells at least in part through activation of adenylate cyclase and protein kinase C signaling pathways and that the ERK1/2 cascade is involved in PACAP and NGF-induced PACAP gene expression, although redundant signaling pathways may also be involved. The present finding showing that PACAP in combination with NGF causes a synergistic increase in PACAP gene expression in PC12 cells supports the idea that PACAP acts as an autocrine regulatory factor.Journal of Neurochemistry 01/2000; 74(2):501 - 507. · 4.06 Impact Factor -
Article: Cloning and characterization of the mouse pituitary adenylate cyclase-activating polypeptide (PACAP) gene
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ABSTRACT: The gene encoding the mouse precursor of pituitary adenylate cyclase-activating polypeptide (PACAP) has been cloned, and its structural organization was determined. Using the 5′-rapid amplification of cDNA ends (RACE) procedure, three types of transcription initiation produced by alternative exon usage of two untranslated alternative exons (exons 1A and 1B) were defined. The PACAP gene spans 6.6 kb of genomic DNA and is composed of six exons including the alternative exons. The signal peptide, PACAP-related peptide and mature 38-amino acid PACAP (PACAP-38) are encoded within exons 2, 4 and 5, respectively. The 5′-flanking region of the PACAP gene contains several sequence motifs homologous to cAMP response element, TPA response element, and growth hormone factor-1 binding site. A dinucleotide repeat sequence is present in an intron. In addition, there are di- and tetranucleotide repeat sequences 2.4 kb and 3.2 kb upstream to the translation start point, respectively. The overall intron–exon organization and the production of the alternate mRNAs of the PACAP gene are markedly similar to those of the growth hormone-releasing hormone (GHRH), supporting the hypothesis that the two genes encoding GHRH or PACAP were originated from a gene duplication. Promoter analysis of the 5′-flanking region of the PACAP gene using a luciferase gene reporter system revealed that the isolated 5′-flanking region has functional promoter activity and is responsible for inducible expression.Gene. -
Article: Genomic Organization and Chromosomal Location of the Mouse Vasoactive Intestinal Polypeptide 1 (VPAC1) Receptor
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ABSTRACT: The gene encoding the mouse vasoactive intestinal polypeptide type 1 (VPAC1) receptor was cloned, and its structural organization was determined. The gene (Vipr1) is more than 16 kb in length and is divided into 13 exons. The 5′-flanking region is highly GC-rich and lacks an apparent TATA box, but contains a CCAAT box, three potential Sp1-binding sites, and two potential AP-2-binding sites. Promoter analysis of the 5′-flanking region ofVipr1using a luciferase gene reporter system revealed that the isolated 5′-flanking region has functional promoter activity. The mouseVipr1gene is encoded by a single gene, which was mapped to the distal region of mouse chromosome 9. This region is syntenic with human chromosome 3p, where the human VPAC1receptor gene has been mapped.Genomics. -
Article: cDNA cloning of a mouse pituitary avenylate cyclase-activating polypeptive receptor
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ABSTRACT: A cDNA clone for mouse pituitary avenylate cyclase-activating polypeptive (PACAP) receptor (PACAP-R) was obtained from the brain using reverse transcription-polymerase chain reaction (RT-PCR). The recombinant PACAP receptor expressed in COS cells bound PACAP with about 1000-times higher affinity than vasoactive intestinal polypeptive (VIP), and PACAP stimulated avenylate cyclase through the cloned PACAP receptor. The mouse PACAP receptor consists of 496 amino acids, contains seven transmembrane segments and has 98.4%, 93.0%, and 92.5% iventity with the rat, bovine, and human PACAP-R, respectively.Biochimica et Biophysica Acta (BBA) - Biomembranes. 1281(2):129-133.
