Adipose Tissue Protocols

1. Front Matter

   Pages i-xiii

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2. Choosing an Adipose Tissue Depot for Sampling

   • Louis Casteilla, Luc Pénicaud, Béatrice Cousin, Denis Calise

   Pages 1-19

3. Morphologic Techniques for the Study of Brown Adipose Tissue and White Adipose Tissue

   • Saverio Cinti, M. Cristina Zingaretti, Raffaella Cancello, Enzo Ceresi, Plinio Ferrara

   Pages 21-51

4. Confocal Microscopy of Adipocytes

   • Daniela Malide

   Pages 53-64

5. Cellularity Measurements

   • Mario DiGirolamo, Jacqueline B. Fine

   Pages 65-75

6. Subcellular Fractionation of Adipocytes and 3T3-L1 Cells

   • Hans-Georg Joost, Annette Schürmann

   Pages 77-82

7. Quantification of Lipid-Related mRNAs by Reverse Transcription-Competitive Polymerase Chain Reaction in Human White Adipose Tissue Biopsies

   • Hubert Vidal

   Pages 83-88

8. Glycogen Synthase Activity in Adipose Tissue

   • Heidi K. Ortmeyer

   Pages 89-96

9. Assays of Lipolytic Enzymes

   • Cecilia Holm, Gunilla Olivecrona, Malin Ottosson

   Pages 97-119

10. Assays of Lipogenic Enzymes

    • Raymond Bazin, Pascal Ferré

    Pages 121-127

11. Assays of Adrenergic Receptors

    • Christian Carpéné

    Pages 129-140

12. Transfection of Adipocytes and Preparation of Nuclear Extracts

    • Isabelle Dugail

    Pages 141-146

13. Assay of Membrane Transport of Long-Chain Fatty Acids by Adipocytes

    • Nada A. Abumrad

    Pages 147-155

14. Assays of Glucose Entry, Glucose Transporter Amount, and Translocation

    • Jean-François Tanti, Mireille Cormont, Thierry Grémeaux, Yannick Le Marchand-Brustel

    Pages 157-165

15. Methods to Study Phosphorylation and Activation of the Hormone-Sensitive Adipocyte Phosphodiesterase Type 3B in Rat Adipocytes

    • Eva Degerman, Svante Resjö, Tova Rahn Landström, Vincent Manganiello

    Pages 167-180

16. Measurements of Glucose Conversion to its Metabolites

    • Mario DiGirolamo

    Pages 181-192

17. Metabolite and Ion Fluxes and Ion Channels in Brown and White Adipocytes

    • Jan Nedergaard

    Pages 193-196

18. Culture of Adipose Tissue and Isolated Adipocytes

    • Susan K. Fried, Naima Moustaid-Moussa

    Pages 197-212

19. Cultures of Adipose Precursor Cells from Brown Adipose Tissue and of Clonal Brown-Adipocyte-Like Cell Lines

    • Barbara Cannon, Jan Nedergaard

    Pages 213-224

20. Cultures of Adipose Precursor Cells and Cells of Clonal Lines from Animal White Adipose Tissue

    • Raymond Négrel, Christian Dani

    Pages 225-237

Adipose tissue is recognized to be exquisitely sensitive to hormone action, and is also now recognized as a secretory and endocrine organ required for reproduction and good health. Adipocytes are “smart” cells able within the tissue to communicate with surrounding cells, but also with various organs, particularly via leptin acting on the central nervous system. Brown adipose tissue (BAT) and white adipose tissue (WAT) are known to be distinct tissues, whereas the heterogeneity of WAT depots is well est- lished. Unfortunately, excess WAT leads to obesity, which is the most common health problem in industrialized countries. Therefore, from both a scientific and a technical point of view, the time has come to create a survey of adipose tissues and their neglected adipocytes. In Adipose Tissue Protocols, I have attempted to gather together chapters from all areas of adipose tissue research—from in vivo to in vitro studies—and to provide methods covering a wide variety of techniques, including the choice of adipose tissue depot and of morphological techniques for the study of BAT and WAT; the isolation, subcellular fractionation, and transfection of adipocytes where the low density of these cells must be taken into account; assays of nutrient and ion fluxes and the metabolic aspects of nutrient uptake; assays of lipid-related enzymes; biopsies and quantification of lipid-related mRNAs; cultures of adipose precursor cells from WAT and BAT of various species, including human tissue; measurements of adipose secretory products; and assessment of WAT metabolism in vivo.

• Université de Nice-Sophia Antipolis, Nice, France

  Gérard Ailhaud

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