Guinea-pig atrial cardioballs

  • M. Bechem
  • F. Pieper
  • L. Pott
Conference paper

Summary

Myocytes from atria of adult guinea-pigs were isolated by means of a previously described enzyme perfusion with some modifications.

The problem of Ca-intolerance of the dispersed cells was circumvented by (i) avoiding cooling of the cells below 25 °C and (ii) increasing the Ca concentration slowly already during the enzyme perfusion.

Isolated atrial myocytes were taken in long-term cell culture. Under this condition they become spherical within about 2 days. The rounded stage (cardioballs), which is found in the cultures for a period of ca. 10 days is highly suited for electrophysiological studies using the different recording configurations of the patch clamp technique, including ‘tight-seal whole-cell recording’ with simultaneous cell dialysis.

Key words

single heart cell cardioball cell culture whole-cell recording 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. I. Altschuld R, Gibb L, Ansel A, Kruger FA, Brierley GP (1980): Calcium tolerance of isolated rat heart cells. J Mol Cell Cardiol 12: 1383–1395CrossRefGoogle Scholar
  2. 2.
    Belardinelli L, Isenberg G (1983): Isolated atrial myocytes: adenosine and acetylcholine increase potassium conductance. Am J Physiol 244: H 734–737Google Scholar
  3. 3.
    Bechem M, Glitsch HG, Pott L (1983a): Properties of an inward rectifying K channel in the membrane of guinea-pig atrial cardioballs. Pflügers Arch 399: 186–193PubMedCrossRefGoogle Scholar
  4. 4.
    Bechem M, Pott L (1984): K-channels activated by loss of intracellular ATP in guinea-pig atrial cardioballs. J Physiol (London) 348: 50 PGoogle Scholar
  5. 5.
    Bechem M, Pott L, Rennebaum H (1983b): Atrial muscle cells from hearts of adult guinea-pigs in culture: a new preparation for cardiac cellular electrophysiology. Europ J Cell Biol 31: 366–369PubMedGoogle Scholar
  6. 6.
    Dow JW, Harding NGL, Powell T (1981): Isolated cardiac myocytes. I. Preparation of adult myocytes and their homology with the intact tissue. Cardiovasc Res 15: 483–514Google Scholar
  7. 7.
    Hamill OP, Marty A, Neher E, Sakmann B, Sigworth F (1981): Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391: 85–100PubMedCrossRefGoogle Scholar
  8. 8.
    Isenberg G, Klöckner U (1982a): Calcium tolerant ventricular myocytes delivered by pre-incubation in a “KB-medium”. Pflügers Arch 395: 6–18PubMedCrossRefGoogle Scholar
  9. 9.
    Isenberg G, Klöckner U (1982b): Calcium currents of isolated bovine ventricular myocytes are fast and of large amplitude. Pflügers Arch 395: 30–41PubMedCrossRefGoogle Scholar
  10. 10.
    Marty A, Neher E (1983): Tight-seal whole-cell recording. In: Sakmann B, Neher E (eds). Single channel recording. Chapter 7, Plenum, New York, 107–122CrossRefGoogle Scholar
  11. IL Piper HM, Probst J, Schwartz P, Hiltter FJ, Spieckermann PG: Culturing of calcium stable adult cardiac myocytes. J Moll Cell Cardiol 14: 397–412Google Scholar
  12. 12.
    Pott L, Bechem M (1984): Properties of muscarinic K channels in guinea-pig atrial cardioballs as studied by ACh injection into patch clamp pipettes. Pflügers Arch (abstr) in pressGoogle Scholar
  13. 13.
    Powell T, Twist VW (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: 327–333PubMedCrossRefGoogle Scholar
  14. 14.
    Powell T, Terrar DA, Twist VW (1980): Electrical properties of individual cells isolated from adult rat ventricular myocardium. J Physiol (London) 302: 131–153Google Scholar
  15. 15.
    Sakmann B, Trube G (1984): Conductance properties of single inwardly rectifying potassium channels in ventricular cells from guinea-pig heart. J Physiol (London) 347: 641–657Google Scholar
  16. 16.
    Truhe G (1983): Enzymatic dispersion of heart and other tissues. In: Sakmann B, Neher E (eds). Single channel recording, Chapter 4, Plenum, New York, 69–76Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1984

Authors and Affiliations

  • M. Bechem
    • 2
  • F. Pieper
    • 1
  • L. Pott
    • 1
  1. 1.Institut für ZellphysiologieRuhr-Universität BochumBochumGermany
  2. 2.Institut für PharmakologieBayer ForschungszentrumWuppertalGermany

Personalised recommendations