Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 102, Issue 31, Pages 11112-11117Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0500360102
Keywords
evenescent field microscopy; smooth muscle; sparklets
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Funding
- NHLBI NIH HHS [HL 07828, T32 HL007312, T32 HL007828, HL 077115, HL 07312, R01 HL077115] Funding Source: Medline
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Ca2+ influx through L-type Ca2+ channels (LTCCs) influences numerous physiological processes ranging from contraction in muscle and memory in neurons to gene expression in many cell types. However, the spatiotemporal organization of functional LTCCs has been nearly impossible to investigate because of methodological limitations. Here, we examined LTCC function with high temporal and spatial resolution using evanescent field fluorescence microscopy. Surprisingly, we found that LTCCs operated in functionally organized clusters, not necessarily as individual proteins. Furthermore, LTCC function in these clusters does not appear to be controlled by simple stochastic gating but instead by a PKC-dependent switch mechanism. This work suggests that resting intracellular free calcium concentration in arterial myocytes is predominantly controlled by this process in combination with rare voltage-dependent openings of individual LTCCs. We propose that Ca2+ influx via persistent LTCCs may be an important mechanism regulating steady-state local and global Ca2+ signals.
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