title: CFTR chloride channels in human and simian heart.
abstract: OBJECTIVE
  The cAMP-dependent Cl- conductance in heart is believed to be due to cardiac expression of the cystic fibrosis transmembrane conductance regulator (CFTR). While CFTR expressed in rabbit and guinea-pig heart (CFTRcardiac) is an alternatively spliced isoform of the epithelial gene product, little information is known regarding possible expression of CFTR in primate heart. In this study, we examined molecular expression of CFTR in human and simian atrium and ventricle and functional expression of cAMP-dependent Cl- currents in isolated human atrial and simian ventricular cells.
  
  
  METHODS
  The reverse transcription polymerase chain reaction (RT-PCR) was performed on human and simian atrial and ventricular mRNA using primers designed to border regions of the CFTR gene product corresponding to transmembrane segments I-VI (TSI-VI), the first nucleotide binding domain (NBD1), transmembrane segments VII-XII (TSVII-XII), and the large cytoplasmic domain which includes the regulatory (R) domain and NBD1. Functional expression of CFTR Cl- channels in human atrial and simian ventricular myocytes was determined using whole-cell and giant inside-out patch-clamp techniques.
  
  
  RESULTS
  Southern blot analysis of these RT-PCR products demonstrated expression of CFTR transcripts in human and simian atrial and ventricular tissue and revealed a novel pattern of expression compared to most animal species studies: both the exon 5 plus (unspliced) and exon 5 minus (spliced) CFTR transcripts are co-expressed in human and simian atrium and ventricle. Whole-cell experiments demonstrated a Cl- sensitive time-independent background conductance in both human atrial and simian ventricular myocytes that was activated by forskolin (FSK) and insensitive to 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). In inside-out patches utilizing the giant patch technique on human atrial myocytes, unitary Cl- sensitive channels resembling CFTR Cl- channels (approximately 14 pS conductance) were activated by the catalytic subunit of protein kinase A (PKA) in 3/12 patches examined.
  
  
  CONCLUSIONS
  These results clearly demonstrate the molecular expression of CFTR Cl- channels and provide electrophysiological evidence consistent with functional expression of these channels in human atrial and simian ventricular myocardium.
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno 89557-0046, USA.
authors.forename: J D
authors.initials: JD
authors.lastname: Warth
authors.forename: M L
authors.initials: ML
authors.lastname: Collier
authors.forename: P
authors.initials: P
authors.lastname: Hart
authors.forename: Y
authors.initials: Y
authors.lastname: Geary
authors.forename: C H
authors.initials: CH
authors.lastname: Gelband
authors.forename: T
authors.initials: T
authors.lastname: Chapman
authors.forename: B
authors.initials: B
authors.lastname: Horowitz
authors.forename: J R
authors.initials: JR
authors.lastname: Hume
mesh_terms: D000368:Aged; D000818:Animals; D001483:Base Sequence; D018118:Chloride Channels / Q000032:analysis / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006325:Heart Atria / Q000737:chemistry / Q000378:metabolism; D006352:Heart Ventricles / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008252:Macaca fascicularis; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D016133:Polymerase Chain Reaction
chemical_list: C505032:CFTR protein, human; D018118:Chloride Channels; D017931:DNA Primers; D019005:Cystic Fibrosis Transmembrane Conductance Regulator