A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons

This protocol describes using fluorescence recovery after photobleaching (FRAP) of a superecliptic pHluorin (SEP)-diacylglycerol lipase α (DAGLα) to measure membrane-bound DAGLα mobility in dendritic shafts of primary cultured cortical mouse neurons. This could serve as an excellent tool to analyze...

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Detalles Bibliográficos
Autores principales: Yoon, Sehyoun, Penzes, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8783203/
https://www.ncbi.nlm.nih.gov/pubmed/35098165
http://dx.doi.org/10.1016/j.xpro.2021.101118
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author Yoon, Sehyoun
Penzes, Peter
author_facet Yoon, Sehyoun
Penzes, Peter
author_sort Yoon, Sehyoun
collection PubMed
description This protocol describes using fluorescence recovery after photobleaching (FRAP) of a superecliptic pHluorin (SEP)-diacylglycerol lipase α (DAGLα) to measure membrane-bound DAGLα mobility in dendritic shafts of primary cultured cortical mouse neurons. This could serve as an excellent tool to analyze endocannabinoid-mediated synaptic plasticity. We have used this protocol to show that DAGLα surface dynamics play an integral role in regulating the dendritic spine. We also detail how we test the qualities of generated SEP-DAGLα in HEK293T cells by FRAP assay. For complete details on the use and execution of this profile, please refer to Yoon et al. (2021a).
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spelling pubmed-87832032022-01-28 A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons Yoon, Sehyoun Penzes, Peter STAR Protoc Protocol This protocol describes using fluorescence recovery after photobleaching (FRAP) of a superecliptic pHluorin (SEP)-diacylglycerol lipase α (DAGLα) to measure membrane-bound DAGLα mobility in dendritic shafts of primary cultured cortical mouse neurons. This could serve as an excellent tool to analyze endocannabinoid-mediated synaptic plasticity. We have used this protocol to show that DAGLα surface dynamics play an integral role in regulating the dendritic spine. We also detail how we test the qualities of generated SEP-DAGLα in HEK293T cells by FRAP assay. For complete details on the use and execution of this profile, please refer to Yoon et al. (2021a). Elsevier 2022-01-20 /pmc/articles/PMC8783203/ /pubmed/35098165 http://dx.doi.org/10.1016/j.xpro.2021.101118 Text en © 2021 Northwestern University https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Yoon, Sehyoun
Penzes, Peter
A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title_full A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title_fullStr A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title_full_unstemmed A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title_short A fluorescence recovery after photobleaching protocol to measure surface diffusion of DAGLα in primary cultured cortical mouse neurons
title_sort fluorescence recovery after photobleaching protocol to measure surface diffusion of daglα in primary cultured cortical mouse neurons
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8783203/
https://www.ncbi.nlm.nih.gov/pubmed/35098165
http://dx.doi.org/10.1016/j.xpro.2021.101118
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