Molecular and Cellular Biochemistry

, Volume 343, Issue 1–2, pp 107–113 | Cite as

Cell swelling impairs dye coupling in adult rat ventricular myocytes. Cell volume as a regulator of cell communication



The influence of cell swelling on cell communication was investigated in cardiomyocytes isolated from the ventricle of adult rats. Measurements of dye coupling were performed in cell pairs using intracellular dialysis of Lucifer Yellow CH. The pipette was attached to one cell of the pair and after a gig ohm seal was achieved, the membrane was ruptured by a brief suction allowing the dye to diffuse from the pipette into the cell. Fluorescence of the dye in the injected as well as in non-dialyzed cell of the pair was continuously monitored. The results indicate that in cell pairs exposed to hypotonic solution the cell volume was increased by about 60% within 35 min and the dye coupling was significantly reduced by cell swelling. Calculation of gap junction permeability (P j) assuming an the intracellular volume accessible to intracellular diffusion of the dye as 12% of total cell volume, showed an average P j value of 0.16 ± 0.04 × 10−4 cm/s (n = 35) in the control and 0.89 ± 1.1 × 10−5 cm (n = 40) for cells exposed to hypotonic solution (P < 0.05). Similar results were found assuming intracellular volumes accessible to the dye of 20 and 30% of total cell volume, respectively. Cell swelling did not change the rate of intracellular diffusion of the dye. The results which indicate that cell volume is an important regulator of gap junction permeability, have important implications to myocardial ischemia and heart failure as well as to heart pharmacology because changes in cell volume caused by drugs and transmitters can impair cell communication with consequent generation of slow conduction and cardiac arrhythmias.


Cell volume Dye coupling Heart myocardial Ischemia pharmacology 



This work was supported by Grants GM 61838 and G12RR03051 from NIH.


  1. 1.
    Kohl P, Nesbitt AD, Cooper PJ, Lei M (2001) Sudden cardiac death by commotion cordis: role of mechano-electric feedback. Cardiovasc Res 50:280–289CrossRefPubMedGoogle Scholar
  2. 2.
    van Wagoner DR (1993) Mechanosensitive gating of atrial ATP-sensitive potassium channels. Circ Res 72:973–983PubMedGoogle Scholar
  3. 3.
    Janse MJ, Coronel R, Wilms FJG, de Groot JR (2003) Mechanical effects on arrhythmogenesis: from pipette to patient. Progress Biophys Mol Biol 82:187–189CrossRefGoogle Scholar
  4. 4.
    De Mello WC, Gonzalez Castillo M, van Loon P (1983) Intercellular diffusion of Lucifer Yell CH in mammalian cardiac fibers. J Mol Cell Cardiol 15:637–645CrossRefPubMedGoogle Scholar
  5. 5.
    Veenstra R (2002) Biophysics of gap junction channels. In: De Mello WC, Janse M (eds) Heart cell coupling and impulse propagation in health and disease. Kluwer, Norwell, pp 143–183Google Scholar
  6. 6.
    De Mello WC, van Loon P (1987) Influence of cyclic nucleotides on junctional permeability in atrial muscle. J Mol Cell Cardiol 19:83–90CrossRefPubMedGoogle Scholar
  7. 7.
    De Mello WC (1975) Effect of intracellular injection of calcium on cell communication in heart. J Physiol 250:231–236PubMedGoogle Scholar
  8. 8.
    Burt JM, Spray DC (1988) Inotropic agents modulate gap junctional conductance between cardiomyocytes. Am J Physiol 254:H1206–H1210PubMedGoogle Scholar
  9. 9.
    Kleber AG, Riegger CB, Janse MJ (1987) Electrical uncoupling and increase of extracellular resistance after induction of ischemia in isolated, arterially perfused rabbit papillary muscle. Circ Res 61:271–279PubMedGoogle Scholar
  10. 10.
    De Mello WC (2009) Cell swelling, impulse conduction, and cardiac arrhythmias in the failing heart. Opposite effects of angiotensin II and angiotensin (1–7) on cell volume regulation. Mol Cell Biochem 330:211–217CrossRefPubMedGoogle Scholar
  11. 11.
    Powell T, Twist T (1976) A rapid technique for the isolation and purification of adult cardiac muscle cells having respiratory control and a tolerance to calcium. Biochem Biophys Res Commun 72:327–333CrossRefPubMedGoogle Scholar
  12. 12.
    Tanigushi Y, Kokubun S, Noma A, Irisawa H (1981) Spontaneously active cells isolated from sinoatrial and atrioventricular node of the rabbit heart. Jpn J Physiol 31:547–558Google Scholar
  13. 13.
    Boyett MR, Framton JE, Kirby MS (1991) The length, width and volume of isolated rat and ferret ventricular myocytes during twitch contraction and changes in osmotic strength. Exp Physiol 76:259–270PubMedGoogle Scholar
  14. 14.
    Sorenson AL, Tepper D, Sonnenblick EH et al (1985) Size and shape of enzymatically isolated ventricular myocytes from cardiomyopathic hamsters. Cardiov Res 19:793–799CrossRefGoogle Scholar
  15. 15.
    Imanaga I (1987) Cell-to cell coupling studied by diffusional methods in myocardial cells. Experientia 43:108083CrossRefGoogle Scholar
  16. 16.
    Page E, McCallister LP (1973) Studies on the intercalated disk of rat left ventricular cells. J Ultrastr Res 43:388–411CrossRefGoogle Scholar
  17. 17.
    Matter A (1973) A morphometric study on the nexus of rat cardiac muscle. J Cell Biol 56:690–696CrossRefPubMedGoogle Scholar
  18. 18.
    Clemo HF, Stambler BS, Baumgarten CM (1999) Swelling-activated chloride current is persistently activated in ventricular myocytes from dogs with tachycardia-induced congestive heart failure. Circ Res 84:157–165PubMedGoogle Scholar
  19. 19.
    Mills RW, Narayan SM, Andrew D, McCulloch AD (2008) Mechanisms of conduction slowing during myocardial stretch by ventricular volume loading in the rabbit. Am J Physiol Heart Circ Physiol 295:H1270–H1278CrossRefPubMedGoogle Scholar
  20. 20.
    Browe DM, Baumgarten CM (2003) Stretch of beta 1 integrin activates an outwardly rectifying chloride current via FAK and Scr in rabbit ventricular myocytes. J Gen Physiol 122:689–702CrossRefPubMedGoogle Scholar
  21. 21.
    Hume JR, Duan D, Collier ML, Yamazaki J et al (2000) Anion transport in heart. Physiol Rev 80:31–81PubMedGoogle Scholar
  22. 22.
    Wendt-Gallitelli MF, Voigt T, Isenberg G (1993) Microheterogeneity of subsarcolemmal sodium gradients: electron probe microanalysis in guinea pig ventricular myocytes. J Physiol 472:33–34PubMedGoogle Scholar
  23. 23.
    De Mello WC (1976) Influence of the sodium pump on intercellular communication in heart fibres: effect of intracellular injection of sodium ion on electrical coupling. J Physiol 263:171–197PubMedGoogle Scholar
  24. 24.
    De Mello WC (1996) Renin angiotensin system and cell communication in the failing heart. Hypertension 27:1267–1272PubMedGoogle Scholar
  25. 25.
    Sorota S (1995) Tyrosine protein kinase inhibitors prevent activation of cardiac swelling-induced chloride current. Pflügers Arch 431:178–185CrossRefPubMedGoogle Scholar
  26. 26.
    De Mello WC (2004) Heart failure: how important is cellular sequestration? The role of the renin angiotensin aldosterone system. J Mol Cell Cardiol 37:431–438CrossRefPubMedGoogle Scholar
  27. 27.
    Ismailov II, Benos DJ (1995) Effects of phosphorylation on ion channel function. Kidney Int 48:1167–1179CrossRefPubMedGoogle Scholar
  28. 28.
    Laird DW, Puranam KL, Revel JP (1991) Turnover and phosphorylation dynamics of connexin43 gap junction protein in cultured cardiac myocytes. Biochem J 273:67–72PubMedGoogle Scholar
  29. 29.
    Le Guennec JY, Gannier F, Argibay JA, Garnier D (1991) Stretch-induced increase of resting intracellular calcium concentration in guinea pig ventricular myocytes. Exp Physiol 76:975–978PubMedGoogle Scholar
  30. 30.
    Jackson P, Strange K (1993) Volume sensitive anion channels mediate swelling activated inositol and taurine efflux. Am J Physiol 265:C1489–C1500PubMedGoogle Scholar

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© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  1. 1.Department of Pharmacology, School of MedicineUPRSan JuanUSA

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