Real-time detection of acetylcholine release from the human endocrine pancreas

Rayner Rodriguez-Diaz, Robin Dando, Y. Anthony Huang, Per Olof Berggren, Stephen D. Roper, Alejandro Caicedo

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

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 Ca2+ indicator Fura-2 and pressed against isolated human pancreatic islets allow the detection of ACh release. The biosensor approach is simple; the Ca2+ 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.

Original languageEnglish (US)
Pages (from-to)1015-1023
Number of pages9
JournalNature protocols
Volume7
Issue number6
DOIs
StatePublished - Jun 1 2012

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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