Exp Mol Med.
2005 Aug;37(4):311-322.
Representational difference analysis of cDNA identifies novel genes expressed following preconditioning of the heart
- Affiliations
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- 1Trafford Centre for Graduate Medical Education and Research, University of Sussex, Falmer, Brighton BN1 9RY, UK. l.mayne@sussex.ac.uk
- 2The Hatter Institute for Cardiovascular Studies, Centre for Cardiology, University College London Hospital and Medical School, Grafton Way, London WC1E 6DB, UK.
- 3Department of Basic Sciences, The Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK.
- 4Institute of Child Health, University College, London, 30 Guilford Street, London WC1N 1EH, UK.
Abstract
- Preconditioning of the myocardium rapidly induces a number of transcription factors, which are likely to be responsible for a cascade of transcriptional changes underlying the development of delayed adaptation. Identifying these changes provides insight into the molecular pathways elicited by sub-lethal ischaemia and the mechanism leading to delayed adaptation. Genes up-regulated in rabbit myocardium in vivo by ischaemic preconditioning following reperfusion for 2 h, 4 h and 6 h posttreatment were identified by representational difference analysis of cDNA (cDNA. RDA). The area of the left ventricle rendered ischaemic by preconditioning or the equivalent area of sham-treated animals was isolated and cDNA.RDA performed. Three novel genes and six genes with known function where identified, including the TGFbeta receptor interacting protein 1, the alpha isoform of the A subunit of PP2 and the cap binding protein NCBP1. To determine whether expression of these genes correlated with preconditioning per se, expression was measured in myocardium after both ischaemic as well as heat shock induced preconditioning following 2 h, 4 h, and 6 h reperfusion. These genes were induced in rabbit myocardium in vivo by both ischaemia and heat shock, consistent with a fundamental role in the development of delayed adaptation. The well described role of PP2 in modulating the mitogen-activated protein kinase pathway and promoting cell survival is consistent with our previous work, which identified the reperfusion injury salvage kinase pathway in mediating the protective effects of ischaemic preconditioning. Expression of Trip1 and Ncbp1 also implicates TGFbeta signalling pathways and RNA processing and transport in delayed adaptation to stress in the myocardium.
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