Capillary Electrophoresis-High Resolution Mass Spectrometry for Measuring In Vivo Arginine Isotope Incorporation in Alzheimer's Disease Mouse Models.

Abstract

Immune-based metabolic reprogramming of arginine utilization in the brain contributes to the neuronal pathology associated with Alzheimer's disease (AD). To enable our long-term goals of differentiation of AD mouse model genotypes, ages, and sexes based on activity of this pathway, we describe here the novel dosing (using uniformly labeled (13C615N4) arginine) and analysis methods using capillary electrophoresis high-resolution accurate-mass mass spectrometry for isotope tracing of metabolic products of arginine. We developed a pseudoprimed infusion-dosing regimen, using repeated injections, to achieve a steady state of uniformly labeled arginine in 135-195 min post bolus dose. Incorporation of stable isotope labeled carbon and nitrogen from uniformly labeled arginine into a host of downstream metabolites was measured in vivo in mice using serially sampled dried blood spots from the tail. In addition to the dried blood spot time course samples, total isotope incorporation into arginine-related metabolites was measured in the whole brain and plasma after 285 min. Preliminary demonstration of the technique identified differences isotope incorporation in arginine metabolites between male and female mice in a mouse-model of sporadic Alzheimer's disease (APOE4/huNOS2). The technique described herein will permit arginine pathway activity differentiation between mouse genotypes, ages, sexes, or drug treatments in order to elucidate the contribution of this pathway to Alzheimer's disease.

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Published Version (Please cite this version)

10.1021/jasms.1c00055

Publication Info

Adams, Kendra J, Joan G Wilson, David S Millington, M Arthur Moseley, Carol A Colton, J Will Thompson and W Kirby Gottschalk (2021). Capillary Electrophoresis-High Resolution Mass Spectrometry for Measuring In Vivo Arginine Isotope Incorporation in Alzheimer's Disease Mouse Models. Journal of the American Society for Mass Spectrometry. 10.1021/jasms.1c00055 Retrieved from https://hdl.handle.net/10161/23232.

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Scholars@Duke

David Stuart Millington

Professor Emeritus of Pediatrics

Development of new mass spectrometric methods for the analysis of disease-specific metabolites in human physiological fluids for application to the diagnosis of inborn errors of metabolism. The development of automated methods based on tandem mass spectrometry for the analysis of these metabolites in blood spots and urine from neonates, with a view to revolutionizing newborn screening methodology.
Development and application of mass spectrometry based biomarker assays for monitoring treatment and diagnosis of lysosomal storage diseases (LSDs).
Development and application of methods for analysis of drugs used in human clinical trials.
Application of "lab-on-a-chip" technology, based on digital microfluidics, to enzymatic and DNA- based assays used in newborn screening and at the point of care.
Development and application of new methods for studing clucose metabolism both in vivo and in vitro using stable isotope labeling with tandem mass spectrometry.
Development and application of methods for the analysis of urine markers of oxidative stress, using tandem mass spectrometry.

Colton

Carol Anne Colton

Professor in Neurology

Dr. Colton's research has centered on the study of chronic neurodegenerative diseases such as Alzheimer's disease. These diseases have a neuroinflammatory component involving the innate immune system in the CNS. Dr. Colton was among the first scientists to demonstrate that microglia are CNS macrophages and, like other tissue macrophages, respond to injury in the CNS by "killing" invading organisms. Microglia then help to orchestrate the "repair" process after injury. Recent research has focused on the regulation of microglial reactive oxygen species and reactive nitrogen species production as well as other cytoactive macrophage products that are made during the classical and alternative activation states associated with chronic neurodegeneration. Knowledge gained from the basic research program has been translated to the development of novel and extremely useful mouse models of Alzheimer's disease that enable pre-clinical testing of basic mechanisms and of potential therapeutics.



Thompson

J. Will Thompson

Adjunct Assistant Professor in the Department of Pharmacology & Cancer Biology

Dr. Thompson's research focuses on the development and deployment of proteomics and metabolomics mass spectrometry techniques for the analysis of biological systems. He served as the Assistant Director of the Proteomics and Metabolomics Shared Resource in the Duke School of Medicine from 2007-2021. He currently maintains collaborations in metabolomics and proteomics research at Duke, and develops new tools for chemical analysis as a Principal Scientist at 908 Devices in Carrboro, NC.

Gottschalk

William Kirby Gottschalk

Assistant Professor in Neurology

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