A study of the mechanisms of excitation-contraction coupling in frog skeletal muscle based on measurements of [Ca2+ ] transients inside the sarcoplasmic reticulum

Journal of Muscle Research and Cell Motility
J Fernando Olivera, G Pizarro

Abstract

[Ca2+] transients inside the sarcoplasmic reticulum (SR) were recorded in frog skeletal muscle twitch fibers under voltage clamp using the low affinity indicator Mag Fluo 4 (loaded in its AM form) with the purpose of studying the effect on Ca2+ release of extrinsic Ca2+ buffers (i.e. BAPTA) added at high concentration to the myoplasm. When the extrinsic Ca2+ buffer is added to the myoplasm, part of the released Ca2+ binds to it, reducing the Ca2+ signal reported by a myoplasmic indicator. This, in turn, hinders the quantification of the amount of Ca2+ released. Monitoring release by measuring [Ca2+] inside the SR avoids this problem. The application of extrinsic buffers at high concentration reduced the resting [Ca2+] in the SR ([Ca2+]SR) continuously from a starting value close to 400 μM reaching the range of 100 μM in about half an hour. The effect of reducing resting [Ca2+]SR on the Ca2+ permeability of the SR activated by voltage clamp depolarization to 0 mV was studied in cells where the myoplasmic [Ca2+] ([Ca2+]myo) transients were simultaneously recorded with Rhod2. The Ca2+ release flux was calculated from [Ca2+]myo and divided by [Ca2+]SR to obtain the permeability. Peak permeability was significantly reduced, from 0.0...Continue Reading

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Jun 15, 2010·Journal of Muscle Research and Cell Motility·J Fernando Olivera, Gonzalo Pizarro

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Citations

Feb 18, 2021·Journal of Muscle Research and Cell Motility·D Mejía-RaigosaJ C Calderón
Jun 14, 2020·Journal of Theoretical Biology·Gonzalo Pizarro, J Fernando Olivera
Apr 19, 2019·Cell Calcium·Candice KutchukianVincent Jacquemond
Jun 4, 2021·Biochimica Et Biophysica Acta. General Subjects·Andrés F MilánJuan C Calderón

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Methods Mentioned

BETA
biosensor
sedation
dissection

Software Mentioned

Mag
Maxchelator

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