title: A method for the simultaneous analysis of mRNA levels of multiple cardiac ion channels with a multi-probe RNase protection assay.
abstract: OBJECTIVE
  Various pathological conditions can alter cardiac electrophysiological properties not only by physiological responses but also by modifying the gene expression of ion channels (electrical remodelling). To investigate the underlying mechanisms of the latter, electrophysiological alterations would require a simultaneous and comprehensive analysis of the mRNA level of the ion channel genes.
  
  
  RESULTS
  We designed 19 cardiac ion channel cDNA templates to analyse the corresponding mRNAs and classified them into three template sets. Those sets were a voltage-dependent K(+) channel series (rat erg, KvLQT1, Kv4.3, Kv4.2, Kv2.1, Kv1.5, Kv1.4, Kv1.2), an inwardly rectifying K(+) channel series (rat Kir6.2, SUR2A/B, Kir3.4, Kir3.1, Kir2.2, Kir2.1), and an inward cationic ion channel series (rat SCN5A, alpha1C, beta2, alpha2delta2 of cardiac L-type Ca(2+) channel and alpha1G). These cDNA templates were used to synthesize antisense digoxigenin-labelled RNA probes. An amount of the total RNA of 25 microg was adequate to analyse simultaneously the mRNA levels of the ion channel genes with the use of multi-probe RPA, and these three multi-probe template sets enabled us to evaluate the profile of the spatial and temporal transcripts of the cardiac ion channels.
  
  
  CONCLUSIONS
  The newly developed ion channel multi-probe RPA templates provide an aid in the comprehensive analysis of the electrical remodelling of the heart.
authors.affiliation: The First Department of Internal Medicine, Nippon Medical School, 1-1-5, Sendagi, Bunkyo-ku, Tokyo 1138603, Japan. iwasaki@nms.ac.jp
authors.forename: Yu-Ki
authors.initials: YK
authors.lastname: Iwasaki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yamashita
authors.forename: Akiko
authors.initials: A
authors.lastname: Sekiguchi
authors.forename: Seiji
authors.initials: S
authors.lastname: Hatano
authors.forename: Kouichi
authors.initials: K
authors.lastname: Sagara
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Iinuma
authors.forename: Long-Tai
authors.initials: LT
authors.lastname: Fu
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Takao
authors.initials: T
authors.lastname: Katoh
authors.forename: Teruo
authors.initials: T
authors.lastname: Takano
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D066298:In Vitro Techniques; D007473:Ion Channels / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D015336:Molecular Probe Techniques; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012260:Ribonucleases
chemical_list: D007473:Ion Channels; D012333:RNA, Messenger; D012260:Ribonucleases