A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store.
Endoplasmic reticulum calcium homeostasis is critical for cellular functions and is disrupted in diverse pathologies including neurodegeneration and cardiovascular disease. Owing to the high concentration of calcium within the ER, studying this subcellular compartment requires tools that are optimiz...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4599735?pdf=render |
| _version_ | 1819074364737323008 |
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| author | Mark J Henderson Heather A Baldwin Christopher A Werley Stefano Boccardo Leslie R Whitaker Xiaokang Yan Graham T Holt Eric R Schreiter Loren L Looger Adam E Cohen Douglas S Kim Brandon K Harvey |
| author_facet | Mark J Henderson Heather A Baldwin Christopher A Werley Stefano Boccardo Leslie R Whitaker Xiaokang Yan Graham T Holt Eric R Schreiter Loren L Looger Adam E Cohen Douglas S Kim Brandon K Harvey |
| author_sort | Mark J Henderson |
| collection | DOAJ |
| description | Endoplasmic reticulum calcium homeostasis is critical for cellular functions and is disrupted in diverse pathologies including neurodegeneration and cardiovascular disease. Owing to the high concentration of calcium within the ER, studying this subcellular compartment requires tools that are optimized for these conditions. To develop a single-fluorophore genetically encoded calcium indicator for this organelle, we targeted a low affinity variant of GCaMP3 to the ER lumen (GCaMPer (10.19)). A set of viral vectors was constructed to express GCaMPer in human neuroblastoma cells, rat primary cortical neurons, and human induced pluripotent stem cell-derived cardiomyocytes. We observed dynamic changes in GCaMPer (10.19) fluorescence in response to pharmacologic manipulations of the ER calcium store. Additionally, periodic calcium efflux from the ER was observed during spontaneous beating of cardiomyocytes. GCaMPer (10.19) has utility in imaging ER calcium in living cells and providing insight into luminal calcium dynamics under physiologic and pathologic states. |
| first_indexed | 2024-12-21T18:08:21Z |
| format | Article |
| id | doaj.art-161ea35e86594287bc3d469f5096851a |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T18:08:21Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-161ea35e86594287bc3d469f5096851a2022-12-21T18:54:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011010e013927310.1371/journal.pone.0139273A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store.Mark J HendersonHeather A BaldwinChristopher A WerleyStefano BoccardoLeslie R WhitakerXiaokang YanGraham T HoltEric R SchreiterLoren L LoogerAdam E CohenDouglas S KimBrandon K HarveyEndoplasmic reticulum calcium homeostasis is critical for cellular functions and is disrupted in diverse pathologies including neurodegeneration and cardiovascular disease. Owing to the high concentration of calcium within the ER, studying this subcellular compartment requires tools that are optimized for these conditions. To develop a single-fluorophore genetically encoded calcium indicator for this organelle, we targeted a low affinity variant of GCaMP3 to the ER lumen (GCaMPer (10.19)). A set of viral vectors was constructed to express GCaMPer in human neuroblastoma cells, rat primary cortical neurons, and human induced pluripotent stem cell-derived cardiomyocytes. We observed dynamic changes in GCaMPer (10.19) fluorescence in response to pharmacologic manipulations of the ER calcium store. Additionally, periodic calcium efflux from the ER was observed during spontaneous beating of cardiomyocytes. GCaMPer (10.19) has utility in imaging ER calcium in living cells and providing insight into luminal calcium dynamics under physiologic and pathologic states.http://europepmc.org/articles/PMC4599735?pdf=render |
| spellingShingle | Mark J Henderson Heather A Baldwin Christopher A Werley Stefano Boccardo Leslie R Whitaker Xiaokang Yan Graham T Holt Eric R Schreiter Loren L Looger Adam E Cohen Douglas S Kim Brandon K Harvey A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. PLoS ONE |
| title | A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. |
| title_full | A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. |
| title_fullStr | A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. |
| title_full_unstemmed | A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. |
| title_short | A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. |
| title_sort | low affinity gcamp3 variant gcamper for imaging the endoplasmic reticulum calcium store |
| url | http://europepmc.org/articles/PMC4599735?pdf=render |
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