Adipose Tissue Protocols

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Gérard Ailhaud
Springer Science & Business Media, 2001 - Science - 338 pages
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Adipose tissue is now recognized as a widely dispersed secretory organ that exhibits autocrine, paracrine, and endocrine properties, and plays a significant role in obesity, the most common health problem in industrialized countries. In Adipose Tissue Protocols, Gerard Ailhaud and a team of laboratory experts and clinicians describe in step-by-step detail the major techniques needed for the study of adipose tissue and cells. Drawn from both in vivo and in vitro studies, these readily reproducible methods cover a broad range of techniques, including the choice of adipose tissue depot and of morphological techniques for work on both brown adipose tissue (BAT) and white adipose tissue (WAT). Major treatment is accorded the isolation, subcellular fractionation, and transfection of low density adipocytes, as well as the metabolic aspects of nutrient uptake and key assays of nutrient and ion fluxes. Also covered are: biopsies and quantification of lipid-related mRNAs; cultures of adipose precursor cells from WAT and BAT; measurements of adipose secretory products; assessment of WAT metabolism in vivo; and assays of lipid-related enzymes. Innovative and highly practical, Adipose Tissue Protocols offers endocrinologists, physiologists, cell biologists, and pharmacologists a gold-standard collection of proven methods for effective nutritional, physiological, and molecular-level research on adipose tissue."
 

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Contents

Morphologic Techniques for the Study of Brown Adipose Tissue
21
Confocal Microscopy of Adipocytes
53
Cellularity Measurements
65
Subcellular Fractionation of Adipocytes and 3T3L1 Cells
77
Quantification of LipidRelated mRNAs by Reverse
83
Assays of Lipolytic Enzymes
97
Assays of Lipogenic Enzymes
121
Transfection of Adipocytes and Preparation of Nuclear Extracts
141
Metabolite and Ion Fluxes and Ion Channels in Brown
193
Cultures of Adipose Precursor Cells from Brown Adipose Tissue
213
Cultures of Adipose Precursor Cells and Cells
225
Cultures of Human Adipose Precursor Cells
239
Measurements of Peptide and Nonpeptide Secretory Products
249
Assessment of White Adipose Tissue Metabolism
269
Measurement of Adipose Tissue Blood Flow
281
Respiratory and Thermogenic Capacities of Cells
295

Assay of Membrane Transport of LongChain Fatty Acids
147
Assays of Glucose Entry Glucose Transporter Amount
157
Methods to Study Phosphorylation and Activation
167
Measurements of Glucose Conversion to its Metabolites
181
Measurements of White Adipose Tissue Metabolism
305
Immunoassay Measurements of Leptin
323
Index
331
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