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

Sehyoun Yoon*, Peter Penzes

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

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).

Original languageEnglish (US)
Article number101118
JournalSTAR Protocols
Volume3
Issue number1
DOIs
StatePublished - Mar 18 2022

Keywords

  • Cell Biology
  • Cell Membrane
  • Cell culture
  • Cell-based Assays
  • Microscopy
  • Molecular Biology
  • Molecular/Chemical Probes
  • Neuroscience

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Medicine(all)

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