Measurement of rates of cholesterol synthesis using tritiated water

J. M. Dietschy, D. K. Spady

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

Rates of sterol synthesis in various tissues commonly are assessed by assaying levels of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase on isolated microsomes or by measuring the rates of incorporation of various 14C-labeled substrates or [3H]water into cholesterol by whole cell preparations in vitro or by the tissues of the whole animal in vivo. While measurement of activities of HMG-CoA reductase or rates of incorporation of 14C-labeled substrates into cholesterol give useful relative rates of sterol production, neither method yields absolute rates of cholesterol synthesis. The use of [3H]water circumvents the problem of variable and unknown dilution of the specific activity of the precursor pool encountered when 14C-labeled substrates are used and does yield absolute rates of cholesterol synthesis provided that the 3H/C incorporation ratio is known for a particular tissue. In 12 different experimental situations it has been found that from 21 to 27 μg atoms of 3H are incorporated into cholesterol from [3H]water in different tissues of several animal species, so that the 3H/C incorporation ratio is similar under nearly all experimental conditions and varies from 0.78 to 1.00. When administered in vivo [3H]water rapidly equilibrates with intracellular water and is incorporated into sterols within the various organs at rates that are linear with respect to time. From such data it is possible to obtain absolute rates of cholesterol synthesis in the whole animal and in the various organs of the animal. Current data suggest, therefore, that use of [3H]water yields the most accurate rates of cholesterol synthesis both in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)1469-1476
Number of pages8
JournalJournal of Lipid Research
Volume25
Issue number13
StatePublished - 1984

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Cholesterol
Water
Animals
Sterols
Tissue
Substrates
Animal Structures
Hydroxymethylglutaryl CoA Reductases
Coenzyme A
Microsomes
Dilution
Oxidoreductases
Atoms
In Vitro Techniques

ASJC Scopus subject areas

  • Endocrinology

Cite this

Dietschy, J. M., & Spady, D. K. (1984). Measurement of rates of cholesterol synthesis using tritiated water. Journal of Lipid Research, 25(13), 1469-1476.

Measurement of rates of cholesterol synthesis using tritiated water. / Dietschy, J. M.; Spady, D. K.

In: Journal of Lipid Research, Vol. 25, No. 13, 1984, p. 1469-1476.

Research output: Contribution to journalArticle

Dietschy, JM & Spady, DK 1984, 'Measurement of rates of cholesterol synthesis using tritiated water', Journal of Lipid Research, vol. 25, no. 13, pp. 1469-1476.
Dietschy, J. M. ; Spady, D. K. / Measurement of rates of cholesterol synthesis using tritiated water. In: Journal of Lipid Research. 1984 ; Vol. 25, No. 13. pp. 1469-1476.
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