title: [Pacemaker current gene expression of rat mesenchymal stem cells and identification of mesenchymal stem cells expressing human pacemaker current gene (hHCN2)].
abstract: OBJECTIVE
  To study pacemaker current gene expression of mesenchymal stem cells (MSCs) and the electrophysiological property of MSCs expressing human pacemaker current gene.
  
  
  METHODS
  Pacemaker current gene expression of MSCs were studied by real-time quantitative polymerase chain reaction (real-time PCR) and pcDNA3-hHCN2 was transfected with Lipofectin 2000 into MSCs. hHCN2 expression at mRNA and at protein levels in the transfected cells were identified by real-time PCR and Western blot, respectively. The ionic currents of cloned hHCN2 (IhHCN2) were recorded and the current characteristics were studied through the whole-cell patch clamp technique.
  
  
  RESULTS
  mHCN1, mHCN2, mHCN3, mHCN4 represent (0.08+/-0.01)%, (77.16+/-0.03)%, (0.24+/-0.01)%, (22.53+/-0.02)% of total HCN mRNA in MSCs as determined by real-time PCR. Transfected hHCN2 ionic currents were recorded by whole-cell patch clamp and current density-voltage curves were obtained. The threshold for activation of IhHCN2 was approximately -80 mV and this current could be blocked by Cs+ (4 mmol/L). hHCN2 expression in transfected MSCs was detected both at mRNA and protein levels.
  
  
  CONCLUSIONS
  1. mHCN2 and mHCN4 represent the major populations of total HCN mRNA in MSCs. 2. Plasmid pcDNA3-hHCN2 by Lipofectin could be successfully transfected into MSCs with IhHCN2 recorded by whole-cell patch clamp technique, this study provides a basis for future antiarrhythmic gene therapy.
authors.affiliation: Department of Cardiology, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Hong-xia
authors.initials: HX
authors.lastname: Li
authors.forename: Xiang-jun
authors.initials: XJ
authors.lastname: Yang
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Bin
authors.initials: B
authors.lastname: Jiang
authors.forename: Xu-jie
authors.initials: XJ
authors.lastname: Cheng
authors.forename: Tan
authors.initials: T
authors.lastname: Chen
authors.forename: Lian-hua
authors.initials: LH
authors.lastname: Han
authors.forename: Jian-ping
authors.initials: JP
authors.lastname: Song
authors.forename: Zhi-hua
authors.initials: ZH
authors.lastname: Liu
authors.forename: Wen-ping
authors.initials: WP
authors.lastname: Jiang
mesh_terms: D000818:Animals; D054815:Cyclic Nucleotide-Gated Cation Channels; D015870:Gene Expression; D006801:Humans; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D008564:Membrane Potentials / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D015221:Potassium Channels