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Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons
DOI:

12:58 min

November 20, 2012

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Chapters

  • 00:05Title
  • 01:43Preparing Mouse Glial Culture for Neuronal Culture Conditioning
  • 05:10Neuronal Culture
  • 07:51Transfection
  • 09:41TIRF Imaging
  • 11:12Representative TIRF Images of pHluorin-DRD2 Insertion
  • 12:30Conclusion

Summary

Automatic Translation

By tagging the extracellular domains of membrane receptors with superecliptic pHluorin, and by imaging these fusion receptors in cultured mouse neurons, we can directly visualize individual vesicular insertion events of the receptors to the plasma membrane. This technique will be instrumental in elucidating the molecular mechanisms governing receptor insertion to the plasma membrane.

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