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Journal of Lipid Research, Vol 37, 210-220, Copyright © 1996 by Lipid Research, Inc.
X Li, F Catalina, SM Grundy and S Patel
We have developed a procedure to measure the rates of apolipoprotein (apoB)
and triglyceride secretion from the liver of an individual mouse. Using the
well-characterized method of Triton WR-1339 injection to block peripheral
removal of newly secreted VLDL, the rate of triglyceride accumulation is
monitored and at the end of the experimental period, blood is extracted for
quantitative VLDL preparation. ApoB species in isolated VLDL are analyzed
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the mass
of the apoB-48 and apoB-100 species are estimated by Coomassie staining and
laser densitometric scanning, using known quantities of LDL-apoB-100 as
standards. This methodology was applied to measure the rate of apoB
secretion in male and female FVB/N mice and we found that the molar ratio
of newly secreted ApoB-48 to B-100 is 4.6 in the male, and 3.8 in the
female. Measurements of the steady state apoB levels indicate that
liver-derived apoB-48 is cleared from the circulation 7.1 times faster than
B-100 in the male and 4.7 times faster in the female mouse. VLDL apoB-48
fractional turnover is approximately 1800 pools per day in both the male
and female mouse (1814 +/- 139 vs. 1831 +/- 365 respectively, P = 0.92).
ApoB-100 fractional turnover rates are much slower and show a statistically
significant difference between males and females (255 +/- 19 pools per day
vs. 386 +/- 65 pools per day, respectively, P = 0.006). This procedure
provides for quantification of secretory rates of these apo proteins in
vivo, and may be useful for studying the effects of genetic manipulation on
the simultaneous secretion of apoB- 48- and apoB-100-containing VLDL,
afforded by the panoply of transgenic mouse models now available for study,
as well as for effects of diet and drug therapy.
ARTICLES
Method to measure apolipoprotein B-48 and B-100 secretion rates in an individual mouse: evidence for a very rapid turnover of VLDL and preferential removal of B-48- relative to B-100-containing lipoproteins
Center for Human Nutrition, UT Southwestern Medical Center, Dallas, TX 75235-9052, USA.
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