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Confocal Imaging of Neuropeptide Y-pHluorin: A Technique to Visualize Insulin Granule Exocytosis in Intact Murine and Human Islets
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Biologia
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JoVE Journal Biologia
Confocal Imaging of Neuropeptide Y-pHluorin: A Technique to Visualize Insulin Granule Exocytosis in Intact Murine and Human Islets

Confocal Imaging of Neuropeptide Y-pHluorin: A Technique to Visualize Insulin Granule Exocytosis in Intact Murine and Human Islets

DOI:
10.3791/56089-v

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09:41 min

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September 13, 2017

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, , , ,
1Department of Medicine,University of Miami, 2Department of Medicine,University of Toronto

Capítulos

  • 00:05Título
  • 00:48Viral Infection of Intact Isolated Human or Mouse Pancreatic Islets
  • 02:19Reagent Preparation and Experimental Setup
  • 05:23Confocal Imaging
  • 07:14Results: Insulin Granule Exocytosis Imaging Using Neuropeptide Y-pHIuorin
  • 09:09Conclusion

Summary

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We describe a protocol for visualization of insulin exocytosis in intact islets using pHluorin, a pH-sensitive green fluorescent protein. Isolated islets are infected with adenovirus encoding pHluorin coupled to the vesicle cargo neuropeptide Y. This allows for the detection of insulin granule fusion events by confocal microscopy.

Tags

Confocal ImagingNeuropeptide Y-pHluorinInsulin Granule ExocytosisIntact Murine And Human IsletsDiabetesGranule FusionPancreatic IsletsViral InfectionExtracellular SolutionBasal Glucose MediumHyperglycemic Medium

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