Abstract
As the most prototypical G protein-coupled receptor, β-adrenergic receptor (βAR) regulates the pace and strength of heart beating by enhancing and synchronizing L-type channel (LCC) Ca2+ influx, which in turn elicits greater sarcoplasmic reticulum (SR) Ca2+ release flux via ryanodine receptors (RyRs). However, whether and how βAR-protein kinase A (PKA) signaling directly modulates RyR function remains elusive and highly controversial. By using unique single-channel Ca2+ imaging technology, we measured the response of a single RyR Ca2+ release unit, in the form of a Ca2+ spark, to its native trigger, the Ca 2+ sparklet from a single LCC. We found that acute application of the selective βAR agonist isoproterenol (1 μM, ≤20 min) increased triggered spark amplitude in an LCC unitary current-independent manner. The increased ratio of Ca2+ release flux underlying a Ca2+ spark to SR Ca2+ content indicated that βAR stimulation helps to recruit additional RyRs in synchrony. Quantification of sparklet-spark kinetics showed that βAR stimulation synchronized the stochastic latency and increased the fidelity (i.e., chance of hit) of LCC-RyR intermolecular signaling. The RyR modulation was independent of the increased SR Ca 2+ content. The PKA antagonists Rp-8-CPT-cAMP (100 μM) and H89 (10 μM) both eliminated these effects, indicating that βAR acutely modulates RyR activation via the PKA pathway. These results demonstrate unequivocally that RyR activation by a single LCC is accelerated and synchronized during βAR stimulation. This molecular mechanism of sympathetic regulation will permit more fundamental studies of altered βAR effects in cardiovascular diseases.
Original language | English (US) |
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Pages (from-to) | 18028-18033 |
Number of pages | 6 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 106 |
Issue number | 42 |
DOIs | |
State | Published - Oct 20 2009 |
Externally published | Yes |
Keywords
- Calcium signaling
- Excitation-contraction coupling
- Isoproterenol
- Protein kinase A
- β-adrenergic receptor
ASJC Scopus subject areas
- General