{'fraction_non_alphanumeric': 0.047805302042590175, 'fraction_numerical': 0.018252933507170794, 'mean_word_length': 5.933734939759036, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 0, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
Synergistic interactions between Ca2+ entries through L‐type Ca2+ channels and Na+–Ca2+ exchanger in normal and failing rat heart

We used confocal Ca2+ imaging and the patch‐clamp technique to investigate the interplay between Ca2+ entries through L‐type Ca2+ channels (LCCs) and reverse‐mode Na+–Ca2+ exchange (NCX) in activating Ca2+‐induced Ca2+ release (CICR) from the sarcoplasmic reticulum (SR) in cardiac myocytes from normal and failing rat hearts. In normal myocytes exposed to N(6),2′‐O‐dibutyryl adenosine‐3′,5′‐cyclic monophosphate (db‐cAMP, membrane‐permeable form of cAMP), the bell‐shaped voltage dependence of cytosolic Ca2+ transients was dramatically broadened due to activation of SR Ca2+ release at high membrane potentials (30–120 mV). This broadening of Ca2+‐transient voltage dependence could be prevented by KB‐R7943, an inhibitor of the reverse‐mode NCX. Trans‐sarcolemmal Ca2+ entries were measured fluorometrically in myocytes during depolarizing steps to high membrane potentials. The total Ca2+ entry (ΔFTot) was separated into two Ca2+ entry components, LCC‐mediated (ΔFLCC) and NCX‐mediated (ΔFNCX), by exposing the cells to the specific inhibitors of LCCs and reverse‐mode NCX, nifedipine and KB‐R7943, respectively. In the absence of protein kinase A (PKA) stimulation the amplitude of the Ca2+‐inflow signal (ΔFTot) corresponded to the arithmetic sum of the amplitudes of the KB‐R7943‐ and nifedipine‐resistant components (ΔFTot=ΔFLCC+ΔFNCX). PKA activation resulted in significant increases in ΔFTot and ΔFLCC. Paradoxically, ΔFTot became ∼threefold larger than the sum of the ΔFNCX and ΔFLCC components. In myocytes from failing hearts, stimulation of PKA failed to induce a shift in Ca2+ release voltage dependence toward more positive membrane potentials. Although the total and NCX‐mediated Ca2+ entries were increased again, ΔFTot did not significantly exceed the sum of ΔFLCC and ΔFNCX. We conclude that the LCC and NCX Ca2+‐entry pathways interact synergistically to trigger SR Ca2+ release on depolarization to positive membrane potentials in PKA‐stimulated cardiac muscle. In heart failure, this new form of Ca2+ release is diminished and may potentially account for the compromised contractile performance and reduced functional reserve in failing hearts.