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

David Q.H. Wang, Martin C. Carey

Research output: Contribution to journalArticle

75 Scopus citations

Abstract

The rate of intestinal cholesterol (Ch) absorption is an important criterion for quantitation of Ch homeostasis. However, studies in the literature suggest that percent Ch 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 Ch 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 Ch (37 ± 5%) than gallstone-resistant AKR mice (24 ± 4%). We ascertained that sources of error in the literature leading to marked differences in Ch absorption efficiencies between laboratories 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 interexperimental agreement, and the ranges obtained challenge previously held beliefs regarding the spread of intestinal Ch absorption efficiencies in mice. The approaches documented herein provide quantifiable methodologies for exploring genetic mechanisms of Ch absorption, and for investigating the assembly and secretion of chylomicrons, as well as intestinal lipoprotein metabolism in mice.

Original languageEnglish (US)
Pages (from-to)1042-1059
Number of pages18
JournalJournal of Lipid Research
Volume44
Issue number5
DOIs
StatePublished - May 1 2003

Keywords

  • Bile flow
  • Bile salts
  • Chylomicrons
  • Genetics
  • Lecithin
  • Nutrition
  • Radiolabeled sterols
  • Sitostanol

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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