Journal of Biochemistry Advance Access originally published online on June 16, 2008
Journal of Biochemistry 2008 144(4):419-430; doi:10.1093/jb/mvn084
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© 2008 The Japanese Biochemical Society.
Genomic and Expression Analysis of Canine Calcitonin Receptor-stimulating Peptides and Calcitonin/Calcitonin Gene-related Peptide*
Department of Pharmacology, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan
To whom correspondence should be addressed. Tel: +81-6-6833-5012 (ext. 2507), Fax: +81-6-6835-5349, E-mail: minamino{at}ri.ncvc.go.jp
Received May 13, 2008; Accepted June 9, 2008
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Abstract |
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Calcitonin receptor-stimulating peptides (CRSPs) are new members of the calcitonin/calcitonin gene-related peptide (CT/CGRP) family identified in pigs, dogs and other domestic animals, and CRSP-1 is an active ligand for the CT receptor (CT-R). We recently sequenced porcine CRSP genes (Crsps) and found similarity with the CT/CGRP gene (Ct/Cgrp) in sequence and genomic organization. In this study, we identified five Crsps, Crsp-1 to Crsp-5, in dogs. Crsp-1 has five exons with an exon–intron organization identical to that of porcine Crsp-1 or Crsp-2, while Crsp-2 and Crsp-3 have additional CT-2- and CT-3-coding exons like Ct/Cgrp. Crsp-2 was renamed as Ct-2/Crsp-2 because both CRSP-2 and CT-2 mRNAs were tissue-specifically expressed. Crsp-4 and Crsp-5 are presumably generated by retrotransposition. We postulate that Crsps were generated from the gene duplication of Ct/Cgrp, and gained their diversity during mammalian evolution. Among the canine CTs and CRSPs, CRSP-1, CT-1 and CT-2 are active ligands for the CT-R, but CRSP-2 and others are inactive. Canine CRSP-1 and CT-2 are expressed in the central and peripheral systems, while CT-1 is localized in the thyroid gland. These findings indicate that dogs can be used for an experimental model as analysing the physiological roles of the CT/CGRP/CRSP family.
Key Words: calcitonin, calcitonin gene-related peptide, calcitonin receptor, calcitonin receptor-stimulating peptide, genomic organization
Abbreviations:
CGRP, calcitonin gene-related peptide; CL-R, calcitonin-like receptor; CRSP, calcitonin receptor-stimulating peptide; Crsp, CRSP gene; Crsp-5
, pseudogene Crsp-5; CT, calcitonin; CT-coding exon, exon encoding a CT-like peptide; CT-R, CT receptor; C-terminal, carboxy terminal; Ct/Cgrp, CT/CGRP gene; ESE, exonic splice enhancer; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; NCBI, National Center for Biotechnology Information; N-terminal, amino terminal; RACE, rapid amplification of cDNA ends; RAMP, receptor activity-modifying protein; RT–PCR, reverse transcriptase polymerase chain reaction
*The abbreviations of CT, CGRP, CL-R and related words comply with the recommendation by the Committee on Receptor Nomenclature and Drug Classification of the International Union of Basic and Clinical Pharmacology (Poyner, D.R., Sexton, P.M., Marshall, I., Smith, D.M., Quirion, R., Born, W., Muff, R., Fischer, J.A., and Foord, S.M. (2002) International Union of Pharmacology. XXXII. The mammalian calcitonin gene-related peptides, adrenomedullin, amylin, and calcitonin receptors. Pharmacol. Rev. 54, 233–246).
Accession numbers of cDNA sequences determined in this study are CRSP-3, AB294187 [GenBank] ; CRSP-4, AB294188 [GenBank] ; CT-2, AB294189 [GenBank] ; CT-R, AB294191. The accession number of the cDNA sequence predicted in this study is CT-3, AB294190.
Human Ct/
Cgrp is used instead of porcine Ct/Cgrp in this article, because genetic information on porcine Ct/Cgrp is not publicly available.