Potentiation of Contraction as Related to Changes in Free and Total Intracellular Calcium

  • Maria-Fiora Wendt-Gallitelli
  • Gerrit Isenberg
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 311)


In voltage-clamped guinea-pig ventricular myocytes, we studied the potentiation of contraction in dependence on the concentration of intracellular calcium; ionized calcium [Ca2+]c was measured by Indo-1 microfluospectroscopy and total calcium (∑Ca) by electronprobe microanalysis (EPMA). After a 15 min rest period, [Ca2+]c was approx. 90 nM and ∑Ca was below the detection limit (80 µM) in myoplasm (∑Camyo), junctional sarcoplasmic reticulum (∑CaSR) and mitochondria (∑CaMito). Post rest, repetitive clamp steps (1 Hz) potentiated extent and rate of shortening by 300%. In the literature, post-rest potentiation is attributed to the replenishment of SR with releasable calcium; by EPMA the postulated increase in ∑CaSR was measured directly. Post-rest, the peaks of systolic [Ca2+]c transients increased, however only by 40%. In addition, a moderate increase of end-diastolic [Ca2+]c was measured. In an other series of experiments, contraction was potentiated by 800% increase by means of paired voltage-clamp pulses (1 Hz, 36 °C, 2 mM [Ca2+]0). In the potentiated state, end-diastolic [Ca2+]c was 180 nM and ∑Camyo was 0.65 mM. During systole, [Ca2+]c peaked within 20 ms to 950 nM. ∑Camyo rose within 20 ms to 1.4 mM and fell within 40 ms to 1.1 and within 90 ms to 0.8 mM. In contrast, the time course of contraction was slow and peaked at a time (130 ms) when the [Ca2+]c and ∑Camyo transients were finished. We suggest that Ca2+ bound to troponin C (TnC) controls only the onset but not the time course of myofilament interaction. From [Ca2+]c and ∑Camyo we estimated a Ca2+ buffering capacitance of 1.5 mmol ∑Camyo per pCa change, only a fraction of which can be attributed to Ca2+ binding sites on TnC. A model explaining the results requires the assumption of 0.6 mM additional slow, high affinity Ca2+ sites and 2 mM fast, low affinity Ca2+ sites. We discuss that end-diastolic Ca2+ binding to these sites contributes to the potentiation of contraction. Junctional SR. At the end of diastole ∑CaSR was 2.4 mM which is 4 times larger than ∑Camyo. This difference disappeared 20 ms after depolarization (∑CaSR 1.1 mM), within another 20 ms it largely recovered (∑CaSR 2.0 mM). These properties suggest that the junctional SR is a compartment suitable not only for Ca2+ release but also for rapid Ca2+ reuptake. Mitochondria. Paired-pulse potentiation increased end-diastolic ∑CaMito significantly (0.4 mM). During diastolic ∑CaMito followed the changes in ∑Camyo with some delay (peak ∑CaMito of 1.2 mM after 40 ms). According to biochemical literature, we interpret the changes in ∑CaMito to stimulate the activity of dehydrogenases and to adapt the rate of ATP synthesis to the demands of muscle mechanics.


Sarcoplasmic Reticulum Sarcomere Length Paired Pulse Junctional Sarcoplasmic Reticulum Voltage Clamp Protocol 
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Copyright information

© Springer Science+Business Media New York 1992

Authors and Affiliations

  • Maria-Fiora Wendt-Gallitelli
    • 1
  • Gerrit Isenberg
    • 2
  1. 1.Institute of Physiology IIUniversity of TübingenGermany
  2. 2.Department of PhysiologyUniversity of CologneGermany

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