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Am J Physiol Gastrointest Liver Physiol 296: G1151-G1166, 2009. First published April 2, 2009; doi:10.1152/ajpgi.90649.2008
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REVIEW

A guide to Ussing chamber studies of mouse intestine

Lane L. Clarke

Department of Biomedical Sciences, Dalton Cardiovascular Research Center, University of Missouri, Columbia, Missouri

Submitted 10 November 2008 ; accepted in final form 27 March 2009

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.

colon; duodenum; ion transport; nutrient absorption



Address for reprint requests and other correspondence: L. L. Clarke, Univ. of Missouri, 324 D Dalton Cardiovascular Research Ctr., 134 Research Park Dr., Columbia, MO 65211-3300 (e-mail: clarkel{at}missouri.edu)




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Am. J. Physiol. Gastrointest. Liver Physiol.Home page
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Am J Physiol Gastrointest Liver Physiol, October 1, 2009; 297(4): G858 - G859.
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