title: Calcium and mechanically induced potentials in fibroblasts of rat atrium.
abstract: OBJECTIVE
  Electrically non-excitable cardiac fibroblasts in the sino-atrial node region are mechano-sensitive. Rhythmic contraction of adjacent myocardium, or artificial stretch of the tissue, produce a reversible change in the membrane potential: mechanically induced potentials (MIP). Stretch of normal cardiomyocytes can be associated with intracellular calcium changes. The purpose of this study is to use pharmacological interventions to investigate the possibility that stretch-induced Ca2+ entry through ion channels in the sarcolemma and Ca2+ release from internal stores play a role in MIP generation.
  
  
  METHODS
  Isolated spontaneously contracting or artificially stretched preparations of right atrium of rat heart were superfused with physiological solutions. An intracellular floating microelectrode recorded fibroblast MIPs and was also used for injection of current. A dye, Lucifer yellow, applied through the micropipette, identified recording sites. We assessed the role of extracellular Ca2+ using EGTA in the bathing solution. For the role of intracellular Ca2+ in the generation of MIP, several substances that influence [Ca2+]i handling were applied intracellularly by diffusion from the recording microelectrode. These include: BAPTA (to chelate intracellular Ca2+); BHQ, thapsigargin and CPA (to deplete Ca2+ from intracellular stores by inhibition of the endoplasmic reticulum (ER) ATP Ca2+ pump), and caffeine and ryanodine (to induce ER Ca2+ release).
  
  
  RESULTS
  All the pharmacological compounds which were introduced intracellulary, and EGTA applied extracellularly, decreased the amplitude of the MIP to variable degrees. Only thapsigargin induced a bi-phasic response with an initial increase in MIP amplitude, followed by a decrease. MIP duration was reduced by most interventions, exceptions being low extracellular Ca2+, BHQ and ryanodine. Short duration extracellular application of caffeine, which was added to the perfusate as a secondary contractile stimulus, partly restored the MIPs by activation of cardiac contraction. Intracellular current injection, before any intervention, linearly altered both membrane potential (Em) and MIP amplitude (Vm). Application of compounds listed above introduced non-linearity to the Em/Vm relationship.
  
  
  CONCLUSIONS
  We suggest that mechanically induced Ca2+ influx, induced through stretch-activated channels in the plasma membrane, and release of Ca2+ from the endoplasmic reticulum, play key roles in the mechanism of MIP generation. Further, our results demonstrate the existence of functional ryanodine/caffeine-sensitive Ca2+ stores in cardiac fibroblasts.
authors.affiliation: institute of Physiology, Humboldt University, Berlin, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Kiseleva
authors.forename: A
authors.initials: A
authors.lastname: Kamkin
authors.forename: P
authors.initials: P
authors.lastname: Kohl
authors.forename: M J
authors.initials: MJ
authors.lastname: Lab
mesh_terms: D000200:Action Potentials / Q000187:drug effects*; D000818:Animals; D000889:Anti-Arrhythmia Agents / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006325:Heart Atria; D066298:In Vitro Techniques; D008297:Male; D009200:Myocardial Contraction / Q000502:physiology*; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D012433:Ryanodine / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology*; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin
chemical_list: D000889:Anti-Arrhythmia Agents; D002614:Chelating Agents; D013729:Terpenes; D012433:Ryanodine; D002110:Caffeine; D004533:Egtazic Acid; D019284:Thapsigargin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium