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Lateral Diffusion and Exocytosis of Membrane Proteins in Cultured Neurons Assessed using Fluorescence Recovery and Fluorescence-loss Photobleaching
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Lateral Diffusion and Exocytosis of Membrane Proteins in Cultured Neurons Assessed using Fluorescence Recovery and Fluorescence-loss Photobleaching

Lateral Diffusion and Exocytosis of Membrane Proteins in Cultured Neurons Assessed using Fluorescence Recovery and Fluorescence-loss Photobleaching

DOI:
10.3791/3747-v

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11:58 min

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February 29, 2012

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, , ,
1MRC Centre for Synaptic Plasticity,University of Bristol

Chapters

  • 00:05Title
  • 01:33Cell Culture, Viral Transduction, and Protein Expression
  • 02:16FRAP-FLIP Live Cell Imaging: Equipment Set-up and Image Capture Parameters
  • 05:48FRAP-FLIP Live Cell Imaging: Image Acquisition
  • 07:09Data Analysis
  • 09:25Representative FRAP-FLIP Results
  • 11:34Conclusion

Summary

Automatic Translation

Automatic Translation

This report describes the use of live cell imaging and photobleach techniques to determine the surface expression, transport pathways and trafficking kinetics of exogenously expressed, pH-sensitive GFP-tagged proteins at the plasma membrane of neurons.

Tags

Lateral DiffusionExocytosisMembrane ProteinsCultured NeuronsFluorescence RecoveryFluorescence-loss PhotobleachingTraffickingReceptorsIon ChannelsSynaptic ProteinsSuper-ecliptic PHluorinLive Cell ImagingPlasma Membrane ProteinsProtonationPhoton AbsorptionFluorescence EmissionIntracellular Trafficking EventsLow PH CompartmentsNeutral PH Extracellular EnvironmentPhotodamagePhotobleachingFRAP (fluorescence Recovery After Photobleaching)Protein Dynamics

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