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A guide to Ussing chamber studies of mouse intestine.

Abstract

The Ussing chamber provides a physiological system to measure the transport of ions, nutrients, and drugs across various epithelial tissues. One of the most studied epithelia is the intestine, which has provided several landmark discoveries regarding the mechanisms of ion transport processes. Adaptation of this method to mouse intestine adds the dimension of investigating genetic loss or gain of function as a means to identify proteins or processes affecting transepithelial transport. In this review, the principles underlying the use of Ussing chambers are outlined including limitations and advantages of the technique. With an emphasis on mouse intestinal preparations, the review covers chamber design, commercial equipment sources, tissue preparation, step-by-step instruction for operation, troubleshooting, and examples of interpretation difficulties. Specialized uses of the Ussing chamber such as the pH stat technique to measure transepithelial bicarbonate secretion and isotopic flux methods to measure net secretion or absorption of substrates are discussed in detail, and examples are given for the adaptation of Ussing chamber principles to other measurement systems. The purpose of the review is to provide a practical guide for investigators who are new to the Ussing chamber method.

Authors+Show Affiliations

Univ. of Missouri, 324 D Dalton Cardiovascular Research Ctr., 134 Research Park Dr., Columbia, MO 65211-3300, USA. clarkel@missouri.edu

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Review

Language

eng

PubMed ID

19342508

Citation

Clarke, Lane L.. "A Guide to Ussing Chamber Studies of Mouse Intestine." American Journal of Physiology. Gastrointestinal and Liver Physiology, vol. 296, no. 6, 2009, pp. G1151-66.
Clarke LL. A guide to Ussing chamber studies of mouse intestine. Am J Physiol Gastrointest Liver Physiol. 2009;296(6):G1151-66.
Clarke, L. L. (2009). A guide to Ussing chamber studies of mouse intestine. American Journal of Physiology. Gastrointestinal and Liver Physiology, 296(6), pp. G1151-66. doi:10.1152/ajpgi.90649.2008.
Clarke LL. A Guide to Ussing Chamber Studies of Mouse Intestine. Am J Physiol Gastrointest Liver Physiol. 2009;296(6):G1151-66. PubMed PMID: 19342508.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A guide to Ussing chamber studies of mouse intestine. A1 - Clarke,Lane L, Y1 - 2009/04/02/ PY - 2009/4/4/entrez PY - 2009/4/4/pubmed PY - 2009/7/31/medline SP - G1151 EP - 66 JF - American journal of physiology. Gastrointestinal and liver physiology JO - Am. J. Physiol. Gastrointest. Liver Physiol. VL - 296 IS - 6 N2 - The Ussing chamber provides a physiological system to measure the transport of ions, nutrients, and drugs across various epithelial tissues. One of the most studied epithelia is the intestine, which has provided several landmark discoveries regarding the mechanisms of ion transport processes. Adaptation of this method to mouse intestine adds the dimension of investigating genetic loss or gain of function as a means to identify proteins or processes affecting transepithelial transport. In this review, the principles underlying the use of Ussing chambers are outlined including limitations and advantages of the technique. With an emphasis on mouse intestinal preparations, the review covers chamber design, commercial equipment sources, tissue preparation, step-by-step instruction for operation, troubleshooting, and examples of interpretation difficulties. Specialized uses of the Ussing chamber such as the pH stat technique to measure transepithelial bicarbonate secretion and isotopic flux methods to measure net secretion or absorption of substrates are discussed in detail, and examples are given for the adaptation of Ussing chamber principles to other measurement systems. The purpose of the review is to provide a practical guide for investigators who are new to the Ussing chamber method. SN - 0193-1857 UR - https://www.unboundmedicine.com/medline/citation/19342508/A_guide_to_Ussing_chamber_studies_of_mouse_intestine_ L2 - http://www.physiology.org/doi/full/10.1152/ajpgi.90649.2008?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -