The effects of strontium ions, Sr2+, on Ca2+‐dependent feedback mechanisms during excitation‐contraction coupling were examined in voltage‐clamped rat ventricular myocytes in which intracellular [Ca2+] and [Sr2+] were monitored with the fluorescent indicator, indo‐1.
Voltage clamp depolarizations and caffeine applications during superfusion in Ca2+‐free, Sr2+‐containing solutions were employed to exchange intracellular Ca2+ with Sr2+. Myocytes were loaded with Sr2+ by applying voltage clamp depolarizations during superfusion in Na+‐free, Sr2+‐containing solutions.
Caffeine applications produced large fluorescence transients in Sr2+‐loaded cells. Thus, Sr2+ could be sequestered and released from the sarcoplasmic reticulum.
Ca2+ influx, but not Sr2+ influx, via sarcolemmal Ca2+ channels evoked ryanodine‐sensitive fluorescence transients in Sr2+‐loaded cells. These results demonstrated that Ca2+ influx‐induced Sr2+ release (CISR) from the sarcoplasmic reticulum occurred in these experiments, even though Sr2+ influx‐induced Sr2+ release was not observed.
The amplitude of the Ca2+ influx‐induced fluorescence transient was 17 ± 1% of the caffeine‐induced transient (n= 5 cells), an indication that fractional utilization of Sr2+ sequestered in the sarcoplasmic reticulum during CISR was low.
With increased Sr2+ loading, the amplitude of Ca2+ influx‐ and caffeine‐induced fluorescence transients increased, but fractional utilization of sarcoplasmic reticulum divalent cation stores was independent of the degree of Sr2+ loading. These data suggest that Ca2+ influx directly activated the release of divalent cations from the sarcoplasmic reticulum, but mechanisms promoting positive feedback of Sr2+ release were minimal during CISR.
By comparison, in Ca2+‐loaded myocytes, Ca2+ influx‐induced Ca2+ release (CICR) utilized a greater fraction of caffeine‐releasable stores than CISR. Fractional utilization of Ca2+ stores during CICR increased with the degree of Ca2+ loading.
Taken together, these results suggest that Ca2+‐dependent feedback mechanisms play a major role in determining the extent of sarcoplasmic reticulum Ca2+ release during cardiac excitation‐contraction coupling under a wide range of conditions.