Summary
Adipose tissue development undergoes remodeling in terms of newly synthesized cells (hyperplasia) and newly synthesized lipids that accumulate in adipocytes (hypertrophy). Synthesis and/or breakdown rates of adipose cells and lipids follow a continuous and dynamic pattern, e.g., during obesity development. This chapter describes a unique in vivo method to measure the dynamics of adipose tissue growth using 2H2O labeling and mass spectrometry analyses. The approach uses 2H2O as a metabolic tracer to label the adipose tissue components such as the triglycerides (TG), the fatty acids, and the genomic DNA. Deuterium from 2H2O incorporates in the C–H bonds of glycerol moiety of TG through glyceroneogenesis as well as in palmitate moiety through de novo lipogenesis (DNL). Deuterium also incorporates into DNA through the de novo nucleoside synthesis pathway. The labeled water, 2H2O, is administrated intraperitoneally and/or orally in rodents or in humans for a defined duration and biopsies are collected at the end of the labeling period. We describe the procedure to extract, isolate, and purify the adipose components (TG-glycerol, TG-palmitate, and genomic DNA) and the derivation procedure to analyze the isotopic 2H-enrichment of these components by gas chromatography/mass spectrometry. The calculation principles are described to obtain the fractional and absolute synthesis rates of TG, of DNL, and of DNA measured in the adipose tissues. The method is nonradioactive, nonhazardous, accurate, reproducible, and very sensitive. We present recent in vivo data on the ontogeny of adipose tissue growth dynamics in young and adult obese Zucker rats compared with lean Zucker rats.
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Acknowledgment
We particularly thank Marc Hellerstein, Katherine Macé, Corinne Ammon-Zufferey, and Mireille Moser for their collaboration to the success of the present work. We thank the animal house staff. The present work was co financed by KineMed Inc and Nestec Ltd.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Pouteau, E., Beysen, C., Saad, N., Turner, S. (2009). Dynamics of Adipose Tissue Development by 2H2O Labeling. In: Armstrong, D. (eds) Lipidomics. Methods in Molecular Biology, vol 579. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-322-0_17
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DOI: https://doi.org/10.1007/978-1-60761-322-0_17
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