Measurement of free Ca2+ changes and total Ca2+ release in a single striated muscle fibre using the fluorescent indicator quin 2

Biochem Biophys Res Commun. 1985 May 16;128(3):1180-9. doi: 10.1016/0006-291x(85)91065-4.

Abstract

The fluorescent Ca2+ indicator, quin 2, has been used in isolated striated muscle fibres. There is a distinct quin 2 fluorescence peak at lambda 500 nm upon excitation at lambda 339 nm after axial injection of the potassium salt of quin 2, pH 7.1. Single voltage-clamp or current clamp electrical stimulation resulted in a distinct transient change in the fluorescence at lambda 500 nm which was not observed at lambda 400 nm, the peak of the fibre autofluorescence. Ca2+ buffering is marked at high quin 2 concentrations (greater than or equal to 400 microM) producing a slow decay of force and fluorescence. At lower concentrations (8-30 microM) of quin, the decay of force is within the range observed in non-injected control fibres. A Kd of 457 nM at 5 mM free Mg2+ suggests an upper resting free Ca2+ concentration of 310 nM at 12 degrees C.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoquinolines*
  • Animals
  • Calcium / metabolism*
  • Electric Stimulation
  • Fluorescent Dyes
  • In Vitro Techniques
  • Muscles / metabolism*
  • Sarcoplasmic Reticulum / metabolism
  • Thoracica / metabolism

Substances

  • Aminoquinolines
  • Fluorescent Dyes
  • Quin2
  • Calcium