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

Xiaohua Li, Fernando Catalina, Scott M Grundy, Shailesh Patel

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88 Citations (Scopus)

Abstract

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 transgcnic mouse models now available for study, as well as for effects of diet and drug therapy.

Original languageEnglish (US)
Pages (from-to)210-220
Number of pages11
JournalJournal of Lipid Research
Volume37
Issue number1
StatePublished - Jan 1996

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Apolipoprotein B-48
Apolipoprotein B-100
Lipoproteins
Apolipoproteins B
Liver
Secretory Rate
Diet Therapy
Drug therapy
Apolipoproteins
Nutrition
Electrophoresis
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Triglycerides
Lasers
Blood
Staining and Labeling
Scanning
Drug Therapy
Injections

Keywords

  • Gel electrophoresis
  • Hepatic secretion
  • Laser densitometry
  • mRNA editing
  • Triton

ASJC Scopus subject areas

  • Endocrinology

Cite this

@article{c4c01fd743f147768f19208dfd35dfd1,
title = "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",
abstract = "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 transgcnic mouse models now available for study, as well as for effects of diet and drug therapy.",
keywords = "Gel electrophoresis, Hepatic secretion, Laser densitometry, mRNA editing, Triton",
author = "Xiaohua Li and Fernando Catalina and Grundy, {Scott M} and Shailesh Patel",
year = "1996",
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language = "English (US)",
volume = "37",
pages = "210--220",
journal = "Journal of Lipid Research",
issn = "0022-2275",
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TY - JOUR

T1 - Method to measure apolipoprotein B-48 and B-100 secretion rates in an individual mouse

T2 - Evidence for a very rapid turnover of VLDL and preferential removal of B-48- Relative to B-100-containing lipoproteins

AU - Li, Xiaohua

AU - Catalina, Fernando

AU - Grundy, Scott M

AU - Patel, Shailesh

PY - 1996/1

Y1 - 1996/1

N2 - 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 transgcnic mouse models now available for study, as well as for effects of diet and drug therapy.

AB - 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 transgcnic mouse models now available for study, as well as for effects of diet and drug therapy.

KW - Gel electrophoresis

KW - Hepatic secretion

KW - Laser densitometry

KW - mRNA editing

KW - Triton

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