Hurley, M., Shah, S., Sheard, T. orcid.org/0000-0003-4940-3188 et al. (4 more authors) (2024) Super-resolution analysis of the origins of the elementary events of ER calcium release in dorsal root ganglion neurons. Cells, 13 (1). 38. ISSN 2073-4409
Abstract
Coordinated events of calcium (Ca2+) released from the endoplasmic reticulum (ER) are key second messengers in excitable cells. In pain-sensing dorsal root ganglion (DRG) neurons, these events can be observed as Ca2+ sparks, produced by a combination of ryanodine receptors (RyR) and inositol 1,4,5- triphosphate receptors (IP3R1). These microscopic signals offer the neuronal cells with a possible means of modulating the sub-plasmalemmal Ca2+ handling, initiating vesicular exocytosis. With super-resolution dSTORM and expansion microscopies, we visualised the nanoscale distributions of both RyR and IP3R1 that featured loosely organised clusters in the sub-plasmalemmal regions of cultured, rat DRG somata. We adapted a novel correlative microscopy protocol to examine the nanoscale patterns of RyR and IP3R1 in the locality of each Ca2+ spark. We found that most subplasmalemmal sparks correlated with relatively small groups of RyR whilst larger sparks were often associated with larger groups of IP3R1. These data also showed spontaneous Ca2+ sparks in < 30% of the sub-plasmalemmal cell area but consisted of both these channel species at a 3.8-5 times higher density than in non-active regions of the cell. Taken together, these observations reveal distinct patterns and length-scales of RyR and IP3R1 co-clustering at contact sites between the ER and the surface plasmalemma that encode the positions and the quantity of Ca2+ released at each Ca2+ spark.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
Keywords: | Dorsal Root Ganglion; correlative microscopy; dSTORM; expansion microscopy; ryanodine receptor-3; inositol 1,4,5-trisphosphate receptor; calcium signalling |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 22 Dec 2023 15:23 |
Last Modified: | 04 Jan 2024 12:23 |
Status: | Published |
Publisher: | MDPI AG |
Refereed: | Yes |
Identification Number: | 10.3390/cells13010038 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:206902 |