Quantifying apoprotein synthesis in rodents: Coupling LC-MS/MS analyses with the administration of labeled water

Haihong Zhou, Wenyu Li, Sheng Ping Wang, Vivienne Mendoza, Raymond Rosa, James Hubert, Kithsiri Herath, Theresa McLaughlin, Rory J. Rohm, Michael E. Lassman, Kenny K. Wong, Douglas G. Johns, Stephen F. Previs, Brian K. Hubbard, Thomas P. Roddy

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Stable isotope tracer studies of apoprotein flux in rodent models present difficulties as they require working with small volumes of plasma. We demonstrate the ability to measure apoprotein flux by administering either 2H- or 18O-labeled water to mice and then subjecting samples to LC-MS/MS analyses; we were able to simultaneously determine the labeling of several proteolytic peptides representing multiple apoproteins. Consistent with relative differences reported in the literature regarding apoprotein flux in humans, we found that the fractional synthetic rate of apoB is greater than apoA1 in mice. In addition, the method is suitable for quantifying acute changes in protein flux: we observed a stimulation of apoB production in mice following an intravenous injection of Intralipid and a decrease in apoB production in mice treated with an inhibitor of microsomal triglyceride transfer protein. In summary, we demonstrate a high-throughput method for studying apoprotein kinetics in rodent models. Although notable differences exist between lipoprotein profiles that are observed in rodents and humans, we expect that the method reported here has merit in studies of dyslipidemia as i) rodent models can be used to probe target engagement in cases where one aims to modulate apoprotein production and ii) the approach should be adaptable to studies in humans.

Original languageEnglish
Pages (from-to)1223-1231
Number of pages9
JournalJournal of Lipid Research
Volume53
Issue number6
DOIs
StatePublished - Jun 2012

Keywords

  • Cardiovascular disease
  • Dyslipidemia
  • Kinetic biomarker
  • Liquid chromatographytandem mass spectrometry
  • Stable isotopes

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