Abstract
To investigate whether activity of the sarcolemmal Na pump modulates
the influence of sodium current on excitation-contraction (E-C) coupling,
we measured Ca$^2+$(i) transients (fluo-3) in single voltage-clamped
mouse ventricular myocytes (Na$^+$(pip) = 15 or 0 mM) when
the Na pump was activated (4.4 mM K$^+$(o)) and during abrupt
inhibition of the pump by exposure to 0 K with a rapid solution-switcher
device. After induction of steady state Ca$^2+$(i) transients
by conditioning voltage pulses (0.25 Hz), inhibition of the Na pump
for 1.5 s immediately before and continuing during a voltage pulse
(200 ms, -80 to 0 mV) caused a significant increase (15 +/- 2\%;
n = 16; p < 0.01) in peak systolic Ca$^2+$(i) when Na$^+$(pip)
was 15 mM. In the absence of sodium current (I(Na), which was blocked
by 60 microM tetrodotoxin (TTX)), inhibition of the Na pump immediately
before and during a voltage pulse did not result in an increase in
peak systolic Ca$^2+$(i). Abrupt blockade of I(Na) during a
single test pulse with TTX caused a slight decrease in peak Ca$^2+$(i),
whether the pump was active (9\%) or inhibited (10\%). With the reverse-mode
Na/Ca exchange inhibited by KB-R 7943, inhibition of the Na pump
failed to increase the magnitude of the peak systolic Ca$^2+$(i)
(4 +/- 1\%; p = NS) when Na$^+$(pip) was 15 mM. When Na$^+$(pip)
was 0 mM, the amplitude of the peak systolic Ca$^2+$(i) was
not altered by abrupt inhibition of the Na pump immediately before
and during a voltage pulse. These findings in adult mouse ventricular
myocytes indicate the Na pump can modulate the influence of I(Na)
on E-C coupling in a single beat and provide additional evidence
for the existence of Na fuzzy space, where Na$^+$ can significantly
modulate Ca$^2+$ influx via reverse Na/Ca exchange.
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