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A Method for Mouse Pancreatic Islet Isolation and Intracellular cAMP Determination

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Published on: June 25, 2014

Expression and function of cannabinoid receptors in mouse islets.

Chen Li¹, James E Bowe, Peter M Jones

¹Diabetes Research Group, King's College London, London, UK.

|November 25, 2010

Summary

The endocannabinoid system influences energy balance. Cannabinoid receptors (CB1 and CB2) in mouse islet beta-cells enhance insulin secretion, aiding glucose homeostasis.

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Area of Science:

Endocrinology
Metabolism
Cell Biology

Background:

The endocannabinoid system regulates energy homeostasis.
Cannabinoid receptors (CB1, CB2) have central and peripheral effects.
Previous studies on islet cannabinoid receptor expression and insulin secretion are conflicting.

Purpose of the Study:

Investigate cannabinoid receptor expression in mouse islet beta- and alpha-cells.
Determine the effects of selective receptor agonists on cellular signaling and insulin secretion.

Main Methods:

RT-PCR and Western blotting for receptor mRNA and protein.
Immunohistochemistry for receptor localization.
Enzyme immunoassay for cyclic AMP.
Microfluorimetry for calcium levels.
Radioimmunoassay for insulin secretion.

Main Results:

Mouse islets express both CB1 and CB2 receptors, localized to beta-cells.
Activation of CB1 and CB2 receptors decreased cyclic AMP and increased calcium.
Receptor activation potentiated glucose-stimulated insulin secretion.

Conclusions:

Islet CB1 and CB2 receptors are present in beta-cells.
Activation of these receptors enhances insulin secretion.
Endocannabinoid signaling may regulate insulin secretion for glucose homeostasis.