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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Main Author: Koh, Charmay Pearl Lin
Other Authors: Ayumu Tashiro
Format: Thesis-Doctor of Philosophy
Language:English
Published: Nanyang Technological University 2023
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Online Access:https://hdl.handle.net/10356/165153
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spelling sg-ntu-dr.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
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
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
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.
author2 Ayumu Tashiro
author_facet Ayumu Tashiro
Koh, Charmay Pearl Lin
format Thesis-Doctor of Philosophy
author Koh, Charmay Pearl Lin
author_sort Koh, Charmay Pearl Lin
title 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_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_sort aberrant intercellular calcium waves in the ca1 region of the mouse hippocampus injected with gcamp-expressing virus
publisher Nanyang Technological University
publishDate 2023
url https://hdl.handle.net/10356/165153
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