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Papers In Press, published online ahead of print February 16, 2003
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Department of Medicine and Gastroenterology Division, Beth Israel Deaconess Medical Center, Boston, MA 02215
Corresponding Author: dqwang{at}caregroup.harvard.edu
The rate of intestinal cholesterol absorption is an important criterion for quantitation of cholesterol homeostasis. However, studies in the literature suggest that percent cholesterol absorption, measured usually by a fecal dual isotope ratio method, spans a wide range, from 20 to 90% in healthy inbred mice on a chow diet. In the present study, we adapted four standard methods, one direct (lymph collection) and three indirect (plasma and fecal dual isotope ratio, and sterol balance) measurements of cholesterol absorption and applied them to mice. Our data establish that all methodologies can be valid in mice, with all methods supporting the concept that gallstone-susceptible C57L mice absorb significantly more cholesterol (37±5%) than gallstone-resistant AKR mice (24±4%). We ascertained that sources of error in the literature leading to marked differences in cholesterol absorption efficiencies between authors, relate to a number of technical factors most notably, expertise in mouse surgery, complete solubilization and delivery of radioisotopes, appropriate collection periods for plasma and fecal samples, and total extraction of radioisotopes from feces. We find that all methods provide excellent inter-experimental agreement, and the ranges obtained challenge previously held beliefs regarding the spread of intestinal cholesterol absorption efficiencies in mice. The approaches documented herein, provide quantifiable methodologies for exploring genetic mechanisms of cholesterol absorption, and for investigating the assembly and secretion of chylomicrons, as well as intestinal lipoprotein metabolism.
Revised on February 3, 2003
Accepted on February 4, 2003
Measurement of intestinal cholesterol absorption by plasma and fecal dual isotope ratio, mass balance, and lymph fistula methods in the mouse: An analysis of direct versus indirect methodologies
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