Summary
Lipolysis involves the sequential breakdown of triglycerides into free fatty acids and glycerol. The extent of lipolysis is therefore a key determinant of the energy status of an individual and also dictates insulin resistance. Here, we describe a protocol for estimating lipolysis in murine adipocytes. Glycerol released during the lipolytic reaction is estimated radiometrically to determine the extent of lipolysis within the cell and the data are normalized to cell number.
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References
Kershaw EE, Flier JS (2004) Adipose tissue as an endocrine organ. J Clin Endocrinol Metab 89:2548–2566
Boden G, Lebed B, Schatz M, Homko C, Lemieux S (2001) Effects of acute changes of plasma free fatty acids on intramyocellular fat content and insulin resistance in healthy subjects. Diabetes 50:1612–1617
Brasaemle DL, Rubin B, Harten IA, Gruia-Gray J, Kimmel AR, Londos C (2000) Perilipin A increases triacylglycerol storage by decreasing the rate of triacylglycerol hydrolysis. J Biol Chem 275:38486–38493
Steinberg D, Mayer SE, Khoo JC, Miller EA, Miller RE, Fredholm B, Eichner R (1975) Hormonal regulation of lipase, phosphorylase, and glycogen synthase in adipose tissue. Adv. Cyclic Nucleotide Res 5:549–568
Clifford GM, Londos C, Kraemer FB, Vernon RG, Yeaman GJ (2000) Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes. J Biol Chem 275:5011–5015
Haemmerle G, Lass A, Zimmermann R, Gorkiewicz G, Meyer C, Rozman J, Heldmaier G, Maier R, Theussl C, Eder S, Kratky D, Wagner EF, Klingenspor M, Hoefler G, Zechner R (2006) Defective lipolysis and altered energy metabolism in mice lacking adipose triglyceride lipase. Science 312:734–737
Viswanadha S, Londos C (2006) Optimized conditions for measuring lipolysis in murine primary adipocytes. J Lipid Res 47:1859–1864
Brasaemle DL, Levin DM, Adler-Wailes DC, Londos C (1999) The lipolytic stimulation of 3T3-L1 adipocytes promotes the translocation of cytosolic hormone-sensitive lipase to the surfaces of lipid storage droplets. Biochim Biophys Acta 1483:251–262
Mauriege P, Imbeault P, Prud'homme D, Tremblay A, Nadeau A, Despres JP (2000) Subcutaneous adipose tissue metabolism at menopause: Importance of body fatness and regional fat distribution. J Clin Endocrinol Metab 85:2446–2454
You T, Berman DM, Ryan AS, Nicklas BJ (2006) Effects of hypocaloric diet and exercise training on inflammation and adipocyte lipolysis in obese postmenopausal women. J Clin Endocrinol Metab 89:1739–1746
Klyde BJ, Hirsh J (1979) Isotopic labeling of DNA in rat adipose tissue: evidence for proliferating cells associated with mature adipocytes. J Lipid Res 20:691–704
Acknowledgments
This project was supported by the NIDDK/NIH intramural research program.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Viswanadha, S., Londos, C. (2008). Determination of Lipolysis in Isolated Primary Adipocytes. In: Yang, K. (eds) Adipose Tissue Protocols. Methods in Molecular Biology™, vol 456. Humana Press. https://doi.org/10.1007/978-1-59745-245-8_22
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DOI: https://doi.org/10.1007/978-1-59745-245-8_22
Publisher Name: Humana Press
Print ISBN: 978-1-58829-916-1
Online ISBN: 978-1-59745-245-8
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