Superresolution localization of single functional IP3R channels utilizing Ca2+ flux as a readout.

Biophysical Journal
Steven M WiltgenI Parker

Abstract

The subcellular localization of membrane Ca2+ channels is crucial for their functioning, but is difficult to study because channels may be distributed more closely than the resolution of conventional microscopy is able to detect. We describe a technique, stochastic channel Ca2+ nanoscale resolution (SCCaNR), employing Ca2+-sensitive fluorescent dyes to localize stochastic openings and closings of single Ca2+-permeable channels within <50 nm, and apply it to examine the clustered arrangement of inositol trisphosphate receptor (IP3R) channels underlying local Ca2+ puffs. Fluorescence signals (blips) arising from single functional IP3Rs are almost immotile (diffusion coefficient<0.003 microm2 s(-1)), as are puff sites over prolonged periods, suggesting that the architecture of this signaling system is stable and not subject to rapid, dynamic rearrangement. However, rapid stepwise changes in centroid position of fluorescence are evident within the durations of individual puffs. These apparent movements likely result from asynchronous gating of IP3Rs distributed within clusters that have an overall diameter of approximately 400 nm, indicating that the nanoscale architecture of IP3R clusters is important in shaping local Ca2+ signals...Continue Reading

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Citations

Feb 1, 2012·Journal of Mathematical Biology·Jay Raol, Steven J Cox
May 10, 2011·The Journal of Biological Chemistry·Evangelia Pantazaka, Colin W Taylor
Apr 30, 2014·PloS One·Estefanía PiegariSilvina Ponce Dawson
Mar 1, 2014·Nature Methods·Alex Small, Shane Stahlheber
Oct 29, 2011·Biochimica Et Biophysica Acta·Ana M RossiColin W Taylor
Mar 23, 2012·Biochemical Society Transactions·Taufiq Rahman
Sep 6, 2011·Molecular and Cellular Endocrinology·Colin W Taylor, Philippa Dale
Mar 19, 2013·Biochimica Et Biophysica Acta·Jessica J Li, Christopher M Yip
Jan 1, 2013·Plants·Teresa Vaz MartinsRichard J Morris
Apr 2, 2019·Physical Review Letters·V VoorsluijsG Dupont
Jul 25, 2021·Nature Communications·Nagendra Babu ThillaiappanColin W Taylor
Oct 3, 2013·Cold Spring Harbor Protocols·Colin W TaylorAna M Rossi

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