Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease
Rapid release of calcium from internal stores via ryanodine receptors (RyRs) is one of the fastest types of cytoplasmic second messenger signalling in excitable cells. In the heart, rapid summation of the elementary events of calcium release, 'calcium sparks', determine the contraction of...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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The Royal Society
2023-05-01
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| Series: | Open Biology |
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| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsob.230045 |
| _version_ | 1797821651000229888 |
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| author | Miriam E. Hurley Ed White Thomas M. D. Sheard Derek Steele Izzy Jayasinghe |
| author_facet | Miriam E. Hurley Ed White Thomas M. D. Sheard Derek Steele Izzy Jayasinghe |
| author_sort | Miriam E. Hurley |
| collection | DOAJ |
| description | Rapid release of calcium from internal stores via ryanodine receptors (RyRs) is one of the fastest types of cytoplasmic second messenger signalling in excitable cells. In the heart, rapid summation of the elementary events of calcium release, 'calcium sparks', determine the contraction of the myocardium. We adapted a correlative super-resolution microscopy protocol to correlate sub-plasmalemmal spontaneous calcium sparks in rat right ventricular myocytes with the local nanoscale RyR2 positions. This revealed a steep relationship between the integral of a calcium spark and the sum of the local RyR2s. Segmentation of recurring spark sites showed evidence of repeated and triggered saltatory activation of multiple local RyR2 clusters. In myocytes taken from failing right ventricles, RyR2 clusters themselves showed a dissipated morphology and fragmented (smaller) clusters. They also featured greater heterogeneity in both the spark properties and the relationship between the integral of the calcium spark and the local ensemble of RyR2s. While fragmented (smaller) RyR2 clusters were rarely observed directly underlying the larger sparks or the recurring spark sites, local interrogation of the channel-to-channel distances confirmed a clear link between the positions of each calcium spark and the tight, non-random clustering of the local RyR2 in both healthy and failing ventricles. |
| first_indexed | 2024-03-13T09:55:55Z |
| format | Article |
| id | doaj.art-b38620d58c964e7a883436349b4db563 |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-03-13T09:55:55Z |
| publishDate | 2023-05-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-b38620d58c964e7a883436349b4db5632023-05-23T23:05:04ZengThe Royal SocietyOpen Biology2046-24412023-05-0113510.1098/rsob.230045Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and diseaseMiriam E. Hurley0Ed White1Thomas M. D. Sheard2Derek Steele3Izzy Jayasinghe4School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKSchool of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKSchool of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKSchool of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UKSchool of Biosciences, Faculty of Science, The University of Sheffield, Sheffield S10 2TN, UKRapid release of calcium from internal stores via ryanodine receptors (RyRs) is one of the fastest types of cytoplasmic second messenger signalling in excitable cells. In the heart, rapid summation of the elementary events of calcium release, 'calcium sparks', determine the contraction of the myocardium. We adapted a correlative super-resolution microscopy protocol to correlate sub-plasmalemmal spontaneous calcium sparks in rat right ventricular myocytes with the local nanoscale RyR2 positions. This revealed a steep relationship between the integral of a calcium spark and the sum of the local RyR2s. Segmentation of recurring spark sites showed evidence of repeated and triggered saltatory activation of multiple local RyR2 clusters. In myocytes taken from failing right ventricles, RyR2 clusters themselves showed a dissipated morphology and fragmented (smaller) clusters. They also featured greater heterogeneity in both the spark properties and the relationship between the integral of the calcium spark and the local ensemble of RyR2s. While fragmented (smaller) RyR2 clusters were rarely observed directly underlying the larger sparks or the recurring spark sites, local interrogation of the channel-to-channel distances confirmed a clear link between the positions of each calcium spark and the tight, non-random clustering of the local RyR2 in both healthy and failing ventricles.https://royalsocietypublishing.org/doi/10.1098/rsob.230045ryanodine receptornanodomainscalcium signallingDNA-PAINTcorrelative light microscopy |
| spellingShingle | Miriam E. Hurley Ed White Thomas M. D. Sheard Derek Steele Izzy Jayasinghe Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease Open Biology ryanodine receptor nanodomains calcium signalling DNA-PAINT correlative light microscopy |
| title | Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| title_full | Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| title_fullStr | Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| title_full_unstemmed | Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| title_short | Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| title_sort | correlative super resolution analysis of cardiac calcium sparks and their molecular origins in health and disease |
| topic | ryanodine receptor nanodomains calcium signalling DNA-PAINT correlative light microscopy |
| url | https://royalsocietypublishing.org/doi/10.1098/rsob.230045 |
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