High-resolution measurement and calibration of Ca(2+)-transients using Indo-1 in guinea-pig atrial myocytes under voltage clamp

Cell Calcium. 1991 Apr;12(4):269-77. doi: 10.1016/0143-4160(91)90001-u.

Abstract

Spherical atrial myocytes obtained by enzymatic dispersion of hearts from adult guinea-pigs were loaded with the fluorescent Ca(2+)-indicator Indo-1 via patch-clamp pipettes. The dialysing solution additionally contained citrate (60 mM) as low-affinity ('linear') Ca(2+)-chelating compound in order to slow intracellular Ca(2+)-transients. Changes in Indo-1 fluorescence under voltage-clamp due to Ca(2+)-entry and/or release from the SR were calibrated using an in vivo procedure to determine the limiting fluorescence ratios. Sample recordings will be presented to demonstrate that components of a [Ca2+]i-transient due to entry via L-type Ca(2+)-channels and due to Ca(2+)-release from the SR can be directly visualized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / chemistry
  • Calcium / metabolism*
  • Cells, Cultured
  • Citrates
  • Citric Acid
  • Dialysis
  • Fluorescent Dyes*
  • Guinea Pigs
  • Heart Atria
  • Indoles*
  • Microscopy, Electron
  • Myocardium / cytology
  • Myocardium / metabolism*

Substances

  • Citrates
  • Fluorescent Dyes
  • Indoles
  • Citric Acid
  • indo-1
  • Calcium