AM-loading of fluorescent Ca2+ indicators into intact single fibers of frog muscle

Biophysical Journal
M ZhaoS M Baylor

Abstract

The AM loading of a number of different fluorescent Ca2+ indicators was compared in intact single fibers of frog muscle. Among the 13 indicators studied, loading rates (the average increase in the fiber concentration of indicator per first 60 min of loading) varied approximately 100-fold, from approximately 3 microM/h to >300 microM/h (16 degrees C). Loading rates were strongly dependent on the molecular weight of the AM compounds, with the rate increasing steeply as molecular weight decreased below approximately 850. Properties of delta F/F (the Ca2(+)-related fluorescence signal observed with fiber stimulation) were also measured in AM-loaded fibers and compared with those previously reported for fibers microinjected with indicator. In general, the time course of delta F/F was very similar with AM-loading and microinjection; however, the amplitude of delta F/F was usually smaller with AM-loading. There was a strong correlation between the rate of indicator loading and the value of the parameter f (the ratio of the amplitude of delta F/F in AM-loaded versus microinjected fibers). For indicators with small loading rates (<10 microM/h, N = 5), f values were generally small (< or =0.4, N = 4); whereas with large loading rates (>1...Continue Reading

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Citations

Jul 27, 2011·Journal of Plant Research·Haiyong QuShaoling Zhang
Feb 6, 2009·ACS Chemical Biology·Michael BannwarthKai Johnsson
Sep 30, 2005·Chemical Communications : Chem Comm·Heather A Behanna, Samuel I Stupp
Apr 5, 2012·Cold Spring Harbor Protocols·Stephan D Brenowitz, Wade G Regehr
Feb 3, 1999·Annual Review of Genetics·M A Riley
Nov 30, 2011·The Journal of Physiology·Stephen HollingworthStephen M Baylor
Aug 3, 2002·The Journal of Physiology·Hugh R Matthews, Gordon L Fain
Jun 23, 2012·Cell Calcium·Brian M HagenW Jonathan Lederer
Jul 6, 2010·Progress in Biophysics and Molecular Biology·Stephen M Baylor, Stephen Hollingworth
Apr 1, 2014·Biochimica Et Biophysica Acta·Martin OheimMayeul Collot
Jul 15, 2015·The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology·Jeong Hoon LeeChae Hun Leem
Jul 6, 2014·Cold Spring Harbor Protocols·Stephan D Brenowitz, Wade G Regehr

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