Pacing rate, halothane, and BDM affect fura 2 reporting of [Ca2+]i in intact rat trabeculae

The American Journal of Physiology
Y Jiang, F J Julian

Abstract

Experiments were done on intact trabeculae from rats. Fura 2 in the salt form was microinjected directly into the myoplasm. The experiments were conducted at 30 degrees C, with 2 mM extracellular Ca2+ concentration and pacing at either 0.5 or 5 Hz. The aims were to establish a new method for in vivo calibration of fura 2 and to determine the effect of autofluorescence changes on intracellular Ca2+ concentration ([Ca2+]i) reported by fura 2. Autofluorescence was recorded under optimal conditions for fura 2 fluorescence (emission at 510 nm). By alteration of the oxidation-reduction state, it was shown that NADH is the main component of autofluorescence in heart. An increase in pacing frequency caused a decrease in autofluorescence. Both halothane and 2,3-butanedione monoxime (BDM) at 5-Hz pacing produced a substantial rise in autofluorescence, approaching the levels observed at 0.5-Hz pacing. The values for the dissociation constant (678 nM) and maximum fluorescence ratio of fura 2 for Ca2+ for the in vivo calibration are 3.4 times larger and 2.6 times smaller, respectively, than those found in vitro. Using the parameters obtained in vivo, we found that the diastolic and systolic [Ca2+]i of a twitch at 30 degrees C were 0.2 and 2...Continue Reading

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Citations

Sep 24, 1998·The American Journal of Physiology·Y Jiang, F J Julian
Mar 29, 2001·The Journal of Biological Chemistry·G LambertK Kunzelmann
Jun 18, 2002·The Journal of Biological Chemistry·Jason D KiltsMadan M Kwatra
Aug 12, 2005·Annals of the New York Academy of Sciences·Henk E D J Ter KeursAmir Landesberg
Mar 27, 2001·Annual Review of Pharmacology and Toxicology·S F Steinberg, L L Brunton
Apr 1, 1999·American Journal of Respiratory Cell and Molecular Biology·W KammouniM D Merten
Sep 29, 2000·American Journal of Physiology. Heart and Circulatory Physiology·R C Scaduto, L W Grotyohann
Oct 25, 2003·Journal of Cardiovascular Pharmacology·Jason D KiltsMadan M Kwatra
May 1, 2001·Current Protocols in Toxicology·K K Gunter, T E Gunter
Sep 2, 1998·The American Journal of Physiology·A J BakerS A Camacho
Apr 1, 2008·Annals of the New York Academy of Sciences·Henk E D J Ter KeursAmir Landesberg
Feb 12, 1998·The American Journal of Physiology·Y Jiang, F J Julian
Apr 9, 2008·Progress in Biophysics and Molecular Biology·Henk E D J ter KeursAmir Landesberg

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