| Real-time detection of acetylcholine release from the human endocrine pancreas. | |
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MedLine Citation:
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PMID: 22555241 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Rayner Rodriguez-Diaz; Robin Dando; Y Anthony Huang; Per-Olof Berggren; Stephen D Roper; Alejandro Caicedo |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2012-05-03 |
Journal Detail:
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Title: Nature protocols Volume: 7 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2012 Jun |
Date Detail:
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Created Date: 2012-05-04 Completed Date: 2012-09-07 Revised Date: 2013-02-21 |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1015-23 Citation Subset: IM |
Affiliation:
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Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, Florida, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acetylcholine
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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:
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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:
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0/Fluorescent Dyes; 0/Receptor, Muscarinic M3; 51-84-3/Acetylcholine; 7440-70-2/Calcium; 96314-98-6/Fura-2 |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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