Junctophilin-2 in the nanoscale organisation and functional signalling of ryanodine receptor clusters in cardiomyocytes
Journal of Cell Science
Company of Biologists
© 2016. Published by The Company of Biologists Ltd http://www.biologists.com/user-licence-1-1/
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Signalling nanodomains requiring close contact between the plasma membrane and internal compartments, known as 'junctions', are fast communication hubs within excitable cells such as neurones and muscle. Here we have examined two transgenic murine models probing the role of junctophilin-2, a membrane tethering protein crucial for the formation and molecular organisation of sub-microscopic junctions in ventricular muscle cells of the heart. Quantitative single molecule localisation microscopy showed that junctions in animals producing above-normal levels of junctophilin-2 were enlarged, allowing the re-organisation of the primary functional protein within it, the ryanodine receptor (RyR). Although this change was associated with much enlarged RyR clusters that due to their size should be more excitable, functionally it caused a mild inhibition in the calcium signalling output of the junctions (calcium sparks). Analysis of the single molecule densities of both RyR and junctophilin-2 revealed an ∼3-fold increase in the junctophilin-2 to RyR ratio. This molecular rearrangement is compatible with direct inhibition of RyR opening by junctophilin-2 to intrinsically stabilise the calcium signalling properties of the junction and thus the contractile function of the cell.
This work was supported by Health Research Council of New Zealand (HRC.govt.nz) grant 12/240 and a Human Frontier Science Programme Award to CS. Also supported by an American Heart Association grants (14PRE20490083 to AQ, and 13EIA14560061 to XHTW), National Institutes of Health grants R01-HL089598, R01-HL091947, R01-HL117641, and R41- HL129570 (to XHTW) and UK Royal Society grant RG.IMSB.107729 (to IDJ).
This is the final version of the article. Available from the publisher via the DOI in this record.
Published online October 21, 2016.
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