Document Detail


Real-time detection of acetylcholine release from the human endocrine pancreas.
MedLine Citation:
PMID:  22555241     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Neurons, sensory cells and endocrine cells secrete neurotransmitters and hormones to communicate with other cells and to coordinate organ and system function. Validation that a substance is used as an extracellular signaling molecule by a given cell requires a direct demonstration of its secretion. In this protocol we describe the use of biosensor cells to detect neurotransmitter release from endocrine cells in real-time. Chinese hamster ovary cells expressing the muscarinic acetylcholine (ACh) receptor M3 were used as ACh biosensors to record ACh release from human pancreatic islets. We show how ACh biosensors loaded with the Ca(2+) indicator Fura-2 and pressed against isolated human pancreatic islets allow the detection of ACh release. The biosensor approach is simple; the Ca(2+) signal generated in the biosensor cell reflects the presence (release) of a neurotransmitter. The technique is versatile because biosensor cells expressing a variety of receptors can be used in many applications. The protocol takes ∼3 h.
Authors:
Rayner Rodriguez-Diaz; Robin Dando; Y Anthony Huang; Per-Olof Berggren; Stephen D Roper; Alejandro Caicedo
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-05-03
Journal Detail:
Title:  Nature protocols     Volume:  7     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-05-04     Completed Date:  2012-09-07     Revised Date:  2013-06-25    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1015-23     Citation Subset:  IM    
Affiliation:
Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA.
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MeSH Terms
Descriptor/Qualifier:
Acetylcholine / analysis*
Animals
Biosensing Techniques / methods*
CHO Cells
Calcium / analysis,  metabolism
Cricetinae
Cricetulus
Fluorescent Dyes / analysis
Fura-2 / analysis
Humans
Islets of Langerhans / metabolism*
Receptor, Muscarinic M3 / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
R01 DK084321/DK/NIDDK NIH HHS; R01DC000374/DC/NIDCD NIH HHS; R01DC007630/DC/NIDCD NIH HHS; R01DK084321/DK/NIDDK NIH HHS; R56 DK084321/DK/NIDDK NIH HHS; R56DK084321/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Receptor, Muscarinic M3; 51-84-3/Acetylcholine; 7440-70-2/Calcium; 96314-98-6/Fura-2
Comments/Corrections

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