Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus
The advent of genetically engineered calcium indicators (GECIs) like GCaMP have paved the way for the use of calcium imaging to monitor neuronal activity in neuroscience research. However, accumulating evidence have highlighted undesired phenotypes associated with GCaMP expression. In this study, an...
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| Format: | Thesis-Doctor of Philosophy |
| Language: | English |
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Nanyang Technological University
2023
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| Online Access: | https://hdl.handle.net/10356/165153 |
| _version_ | 1811686673100046336 |
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| author | Koh, Charmay Pearl Lin |
| author2 | Ayumu Tashiro |
| author_facet | Ayumu Tashiro Koh, Charmay Pearl Lin |
| author_sort | Koh, Charmay Pearl Lin |
| collection | NTU |
| description | The advent of genetically engineered calcium indicators (GECIs) like GCaMP have paved the way for the use of calcium imaging to monitor neuronal activity in neuroscience research. However, accumulating evidence have highlighted undesired phenotypes associated with GCaMP expression. In this study, an adeno-associated virus was used to express GCaMP6f in the CA1 region of C57BL6/J mice. Here, we reveal another unprecedented side effect resulting from elevated GCaMP levels – intercellular Ca2+ waves. These waves were characterised, and were also found to be reduced by diazepam and carbenoxolone, but not CPP, and were completely but reversibly blocked by isoflurane anaesthesia. No waves were observed when the original virus used was diluted, suggesting that high GCaMP levels contributed to the appearance of the intercellular Ca2+ waves. Although GCaMP is a very useful genetic tool, researchers should take note of its faults, to ensure the validity of their experimental results. |
| first_indexed | 2024-10-01T05:04:09Z |
| format | Thesis-Doctor of Philosophy |
| id | ntu-10356/165153 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:04:09Z |
| publishDate | 2023 |
| publisher | Nanyang Technological University |
| record_format | dspace |
| spelling | ntu-10356/1651532023-04-04T02:58:00Z Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus Koh, Charmay Pearl Lin Ayumu Tashiro School of Biological Sciences atashiro@ntu.edu.sg Science::Biological sciences The advent of genetically engineered calcium indicators (GECIs) like GCaMP have paved the way for the use of calcium imaging to monitor neuronal activity in neuroscience research. However, accumulating evidence have highlighted undesired phenotypes associated with GCaMP expression. In this study, an adeno-associated virus was used to express GCaMP6f in the CA1 region of C57BL6/J mice. Here, we reveal another unprecedented side effect resulting from elevated GCaMP levels – intercellular Ca2+ waves. These waves were characterised, and were also found to be reduced by diazepam and carbenoxolone, but not CPP, and were completely but reversibly blocked by isoflurane anaesthesia. No waves were observed when the original virus used was diluted, suggesting that high GCaMP levels contributed to the appearance of the intercellular Ca2+ waves. Although GCaMP is a very useful genetic tool, researchers should take note of its faults, to ensure the validity of their experimental results. Doctor of Philosophy 2023-03-16T06:58:43Z 2023-03-16T06:58:43Z 2022 Thesis-Doctor of Philosophy Koh, C. P. L. (2022). Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/165153 https://hdl.handle.net/10356/165153 10.32657/10356/165153 en This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0). application/pdf Nanyang Technological University |
| spellingShingle | Science::Biological sciences Koh, Charmay Pearl Lin Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title | Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title_full | Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title_fullStr | Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title_full_unstemmed | Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title_short | Aberrant intercellular calcium waves in the CA1 region of the mouse hippocampus injected with GCaMP-expressing virus |
| title_sort | aberrant intercellular calcium waves in the ca1 region of the mouse hippocampus injected with gcamp expressing virus |
| topic | Science::Biological sciences |
| url | https://hdl.handle.net/10356/165153 |
| work_keys_str_mv | AT kohcharmaypearllin aberrantintercellularcalciumwavesintheca1regionofthemousehippocampusinjectedwithgcampexpressingvirus |