Identification of PAC1 receptor isoform mRNAs by real-time PCR in rat suprachiasmatic nucleus

Supaporn Ajpru, Angela J. McArthur, Hugh D. Piggins, David Sugden

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The pituitary adenylate cyclase-activating polypeptide (PACAP) has been implicated in the photic resetting of the rodent circadian clock in the suprachiasmatic nucleus (SCN). PACAP can exert its effects via VPAC1, VPAC2 and PAC1 G-protein coupled receptors. PAC1 and VPAC2, but not VPAC1, mRNA is expressed in rat SCN. A variety of PAC1 receptor splice variants have been described showing differences in ligand binding affinity and selectivity, G-protein coupling and ability to activate signal transduction pathways. The present experiments used PCR with isoform specific primers to determine which PAC1 variants are expressed in rat SCN. The PAC1null isoform and a variant containing a single 28-amino acid insert in the third intracellular (IC3) loop (hop1/2) were detected. No other IC3 variants (hip, hip-hop), N-terminal variants (PAC1short, PAC1very short and PAC1(3a)) or the variant differing in transmembrane II and IV (PAC1TM4) were detected in SCN obtained at any time of day. A quantitative real-time PCR assay was established which measured combined expression of the PAC1null/hop variants in rat SCN during a 12:12-h light:dark (L:D) cycle. There was no significant variation of PAC1 mRNA expression (PAC1null+PAC1hop) with time of day. Nor was there a significant difference in the proportion of these two variants with time of day. These results indicate that the phase-dependency of the actions of PACAP on SCN firing and circadian behaviour are not mediated by changes in the level of expression of PAC1 receptor mRNA, nor by phase-dependent expression of PAC1 receptor variants with altered ligand binding, G-protein coupling or signalling characteristics. © 2002 Elsevier Science B.V. All rights reserved.
    Original languageEnglish
    Pages (from-to)29-37
    Number of pages8
    JournalMolecular Brain Research
    Volume105
    Issue number1-2
    DOIs
    Publication statusPublished - 30 Sept 2002

    Keywords

    • Circadian rhythm
    • PAC1 receptor variants
    • PACAP
    • Rat
    • Real-time quantitative PCR
    • Suprachiasmatic nucleus

    Fingerprint

    Dive into the research topics of 'Identification of PAC1 receptor isoform mRNAs by real-time PCR in rat suprachiasmatic nucleus'. Together they form a unique fingerprint.

    Cite this