A role for depolarisation induced calcium entry on the Na-Ca exchange in triggering intracellular calcium release and contraction in rat ventricular myocytes.
OBJECTIVE
The aim was to test whether depolarisation-induced calcium entry on the Na-Ca exchange is able to trigger calcium release from the sarcoplasmic reticulum in rat ventricular myocytes.


METHODS
Myocytes were isolated enzymatically from the left ventricle of the rat heart. Cells were impaled with narrow tipped microelectrodes to minimise intracellular dialysis and maintain normal internal ionic conditions. Cells were voltage clamped, contraction was measured optically, and in some experiments intracellular calcium was measured with Fura-2.


RESULTS
When the fast Na current was inactivated by using a holding potential of -40 mV, Ca entry via L-type Ca channels was expected to be the only mechanism capable of triggering sarcoplasmic reticular Ca release. In this situation, blocking L-type Ca channels should have abolished sarcoplasmic reticular release and the phasic twitch. However, after 2 min exposure to 20 microM nifedipine, which abolished the Ca current (ICa) completely, voltage clamp depolarisation from -40 mV to 0 mV still elicited 41(SEM 8.9)% of the control phasic twitch (n = 22 cells). This shows that there must be another mechanism, besides Ca entry via Ca channels, by which membrane depolarisation can trigger sarcoplasmic reticular release and the phasic twitch. The phasic twitch that remained in the presence of nifedipine increased progressively with the magnitude of step depolarisation, required a functional sarcoplasmic reticulum, was abolished by 5 mM external nickel, and was sensitive to both the Na and Ca transmembrane gradients.


CONCLUSIONS
The voltage dependent sarcolemmal Na-Ca exchange is predicted theoretically to generate a transient Ca entry at the start of a step membrane depolarisation, when membrane potential suddenly becomes more positive than the reversal potential of the Na-Ca exchange. The results of this study indicate that in rat myocytes with normal internal ions, physiological levels of membrane depolarisation generate a sufficient Ca entry on the exchange to trigger sarcoplasmic reticular calcium release and contraction. In the absence of ICa, this mechanism is capable of triggering a calcium release which leads to about 40% of the phasic contraction in cells depolarised from -40 mV to 0 mV. The existence of this sarcoplasmic triggering mechanism may have significance for the normal control of cardiac muscle contraction.
Levi
A J
AJ
Department of Physiology, School of Medical Sciences, University of Bristol, United Kingdom.
Brooksby
P
P
Hancox
J C
JC
D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D002478:Cells, Cultured; D016623:Ion Pumps / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects; D009200:Myocardial Contraction / Q000187:drug effects / Q000502:physiology*; D009206:Myocardium / Q000166:cytology; D009543:Nifedipine / Q000494:pharmacology; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*
D016623:Ion Pumps; D012964:Sodium; D009543:Nifedipine; D002118:Calcium