Skip to main content

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

Molecular and Cellular Biochemistry volume 343, pages 107–113 (2010)Cite this article

Abstract

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.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Kohl P, Nesbitt AD, Cooper PJ, Lei M (2001) Sudden cardiac death by commotion cordis: role of mechano-electric feedback. Cardiovasc Res 50:280–289

    CAS  Article  PubMed  Google Scholar 

  2. van Wagoner DR (1993) Mechanosensitive gating of atrial ATP-sensitive potassium channels. Circ Res 72:973–983

    PubMed  Google Scholar 

  3. Janse MJ, Coronel R, Wilms FJG, de Groot JR (2003) Mechanical effects on arrhythmogenesis: from pipette to patient. Progress Biophys Mol Biol 82:187–189

    Article  Google Scholar 

  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 15:637–645

    Article  PubMed  Google Scholar 

  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 143–183

    Google Scholar 

  6. De Mello WC, van Loon P (1987) Influence of cyclic nucleotides on junctional permeability in atrial muscle. J Mol Cell Cardiol 19:83–90

    Article  PubMed  Google Scholar 

  7. De Mello WC (1975) Effect of intracellular injection of calcium on cell communication in heart. J Physiol 250:231–236

    PubMed  Google Scholar 

  8. Burt JM, Spray DC (1988) Inotropic agents modulate gap junctional conductance between cardiomyocytes. Am J Physiol 254:H1206–H1210

    CAS  PubMed  Google Scholar 

  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 61:271–279

    CAS  PubMed  Google Scholar 

  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 330:211–217

    Article  PubMed  Google Scholar 

  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 72:327–333

    CAS  Article  PubMed  Google Scholar 

  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 31:547–558

    Google Scholar 

  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 76:259–270

    CAS  PubMed  Google Scholar 

  14. Sorenson AL, Tepper D, Sonnenblick EH et al (1985) Size and shape of enzymatically isolated ventricular myocytes from cardiomyopathic hamsters. Cardiov Res 19:793–799

    CAS  Article  Google Scholar 

  15. Imanaga I (1987) Cell-to cell coupling studied by diffusional methods in myocardial cells. Experientia 43:108083

    Article  Google Scholar 

  16. Page E, McCallister LP (1973) Studies on the intercalated disk of rat left ventricular cells. J Ultrastr Res 43:388–411

    CAS  Article  Google Scholar 

  17. Matter A (1973) A morphometric study on the nexus of rat cardiac muscle. J Cell Biol 56:690–696

    CAS  Article  PubMed  Google Scholar 

  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 84:157–165

    CAS  PubMed  Google Scholar 

  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–H1278

    CAS  Article  PubMed  Google Scholar 

  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 122:689–702

    CAS  Article  PubMed  Google Scholar 

  21. Hume JR, Duan D, Collier ML, Yamazaki J et al (2000) Anion transport in heart. Physiol Rev 80:31–81

    CAS  PubMed  Google Scholar 

  22. Wendt-Gallitelli MF, Voigt T, Isenberg G (1993) Microheterogeneity of subsarcolemmal sodium gradients: electron probe microanalysis in guinea pig ventricular myocytes. J Physiol 472:33–34

    CAS  PubMed  Google Scholar 

  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 263:171–197

    PubMed  Google Scholar 

  24. De Mello WC (1996) Renin angiotensin system and cell communication in the failing heart. Hypertension 27:1267–1272

    PubMed  Google Scholar 

  25. Sorota S (1995) Tyrosine protein kinase inhibitors prevent activation of cardiac swelling-induced chloride current. Pflügers Arch 431:178–185

    CAS  Article  PubMed  Google Scholar 

  26. De Mello WC (2004) Heart failure: how important is cellular sequestration? The role of the renin angiotensin aldosterone system. J Mol Cell Cardiol 37:431–438

    Article  PubMed  Google Scholar 

  27. Ismailov II, Benos DJ (1995) Effects of phosphorylation on ion channel function. Kidney Int 48:1167–1179

    CAS  Article  PubMed  Google Scholar 

  28. Laird DW, Puranam KL, Revel JP (1991) Turnover and phosphorylation dynamics of connexin43 gap junction protein in cultured cardiac myocytes. Biochem J 273:67–72

    CAS  PubMed  Google Scholar 

  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 76:975–978

    PubMed  Google Scholar 

  30. Jackson P, Strange K (1993) Volume sensitive anion channels mediate swelling activated inositol and taurine efflux. Am J Physiol 265:C1489–C1500

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

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

Author information

Affiliations

  1. Department of Pharmacology, School of Medicine, UPR, San Juan, PR, 00936-5067, USA

    Walmor C. De Mello

Authors
  1. Walmor C. De Mello
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Walmor C. De Mello.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

De Mello, W.C. Cell swelling impairs dye coupling in adult rat ventricular myocytes. Cell volume as a regulator of cell communication. Mol Cell Biochem 343, 107–113 (2010). https://doi.org/10.1007/s11010-010-0504-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-010-0504-8

Keywords

  • Cell volume
  • Dye coupling
  • Heart myocardial
  • Ischemia pharmacology