Whole cell electrophysiology of primary cultured murine enterochromaffin cells

Katilyn Knutson; Peter R. Strege; Joyce Li; Andrew B. Leiter; Gianrico Farrugia; Arthur Beyder

doi:10.3791/58112

Whole cell electrophysiology of primary cultured murine enterochromaffin cells

Katilyn Knutson, Peter R. Strege, Joyce Li, Andrew B. Leiter, Gianrico Farrugia, Arthur Beyder

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Enterochromaffin (EC) cells in the gastrointestinal (GI) epithelium constitute the largest subpopulation of enteroendocrine cells. As specialized sensory cells, EC cells sense luminal stimuli and convert them into serotonin (5-hyroxytryptamine, 5-HT) release events. However, the electrophysiology of these cells is poorly understood because they are difficult to culture and to identify. The method presented in this paper outlines primary EC cell cultures optimized for single cell electrophysiology. This protocol utilizes a transgenic cyan fluorescent protein (CFP) reporter to identify mouse EC cells in mixed primary cultures, advancing the approach to obtaining high-quality recordings of whole cell electrophysiology in voltage-and current-clamp modes.

Original language	English (US)
Article number	e58112
Journal	Journal of Visualized Experiments
Volume	2018
Issue number	139
DOIs	https://doi.org/10.3791/58112
State	Published - Sep 26 2018

Keywords

Biology
Current clamp
Enterochromaffin cell
Epithelium
Gastrointestinal
Issue 139
Primary culture
Voltage clamp
Whole cell electrophysiology

ASJC Scopus subject areas

Neuroscience(all)
Chemical Engineering(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.3791/58112

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title = "Whole cell electrophysiology of primary cultured murine enterochromaffin cells",

abstract = "Enterochromaffin (EC) cells in the gastrointestinal (GI) epithelium constitute the largest subpopulation of enteroendocrine cells. As specialized sensory cells, EC cells sense luminal stimuli and convert them into serotonin (5-hyroxytryptamine, 5-HT) release events. However, the electrophysiology of these cells is poorly understood because they are difficult to culture and to identify. The method presented in this paper outlines primary EC cell cultures optimized for single cell electrophysiology. This protocol utilizes a transgenic cyan fluorescent protein (CFP) reporter to identify mouse EC cells in mixed primary cultures, advancing the approach to obtaining high-quality recordings of whole cell electrophysiology in voltage-and current-clamp modes.",

keywords = "Biology, Current clamp, Enterochromaffin cell, Epithelium, Gastrointestinal, Issue 139, Primary culture, Voltage clamp, Whole cell electrophysiology",

author = "Katilyn Knutson and Strege, {Peter R.} and Joyce Li and Leiter, {Andrew B.} and Gianrico Farrugia and Arthur Beyder",

note = "Funding Information: The authors thank Mrs. Lyndsay Busby for administrative assistance and Mr. Robert Highet from Mayo Clinic Division of Engineering for design and 3D printing of the culture dish insert. This work was supported by NIH K08 to AB (DK106456), Pilot and Feasibility Grant to AB from Mayo Clinic Center for Cell Signaling in Gastroenterology (NIH P30DK084567) and 2015 American Gastroenterological Association Research Scholar Award (AGA RSA) to AB and NIH R01 to GF (DK52766). Publisher Copyright: {\textcopyright} 2018, Journal of Visualized Experiments. All rights reserved.",

year = "2018",

month = sep,

day = "26",

doi = "10.3791/58112",

language = "English (US)",

volume = "2018",

journal = "Journal of visualized experiments : JoVE",

issn = "1940-087X",

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T1 - Whole cell electrophysiology of primary cultured murine enterochromaffin cells

AU - Knutson, Katilyn

AU - Strege, Peter R.

AU - Li, Joyce

AU - Leiter, Andrew B.

AU - Farrugia, Gianrico

AU - Beyder, Arthur

N1 - Funding Information: The authors thank Mrs. Lyndsay Busby for administrative assistance and Mr. Robert Highet from Mayo Clinic Division of Engineering for design and 3D printing of the culture dish insert. This work was supported by NIH K08 to AB (DK106456), Pilot and Feasibility Grant to AB from Mayo Clinic Center for Cell Signaling in Gastroenterology (NIH P30DK084567) and 2015 American Gastroenterological Association Research Scholar Award (AGA RSA) to AB and NIH R01 to GF (DK52766). Publisher Copyright: © 2018, Journal of Visualized Experiments. All rights reserved.

PY - 2018/9/26

Y1 - 2018/9/26

N2 - Enterochromaffin (EC) cells in the gastrointestinal (GI) epithelium constitute the largest subpopulation of enteroendocrine cells. As specialized sensory cells, EC cells sense luminal stimuli and convert them into serotonin (5-hyroxytryptamine, 5-HT) release events. However, the electrophysiology of these cells is poorly understood because they are difficult to culture and to identify. The method presented in this paper outlines primary EC cell cultures optimized for single cell electrophysiology. This protocol utilizes a transgenic cyan fluorescent protein (CFP) reporter to identify mouse EC cells in mixed primary cultures, advancing the approach to obtaining high-quality recordings of whole cell electrophysiology in voltage-and current-clamp modes.

AB - Enterochromaffin (EC) cells in the gastrointestinal (GI) epithelium constitute the largest subpopulation of enteroendocrine cells. As specialized sensory cells, EC cells sense luminal stimuli and convert them into serotonin (5-hyroxytryptamine, 5-HT) release events. However, the electrophysiology of these cells is poorly understood because they are difficult to culture and to identify. The method presented in this paper outlines primary EC cell cultures optimized for single cell electrophysiology. This protocol utilizes a transgenic cyan fluorescent protein (CFP) reporter to identify mouse EC cells in mixed primary cultures, advancing the approach to obtaining high-quality recordings of whole cell electrophysiology in voltage-and current-clamp modes.

KW - Biology

KW - Current clamp

KW - Enterochromaffin cell

KW - Epithelium

KW - Gastrointestinal

KW - Issue 139

KW - Primary culture

KW - Voltage clamp

KW - Whole cell electrophysiology

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Whole cell electrophysiology of primary cultured murine enterochromaffin cells

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