Subcutaneous adipose tissue imaging of human obesity reveals two types of adipocyte membranes: Insulin-responsive and -nonresponsive.

Abstract

In adipose tissue, resistance to insulin's ability to increase glucose uptake can be induced by multiple factors, including obesity. Impaired insulin action may take place at different spatial loci at the cellular or subcellular level. To begin to understand the spatial response to insulin in human subcutaneous adipose tissue (hSAT), we developed a quantitative imaging method for activation of a major signaling node in the glucoregulatory insulin signaling pathway. After treatment with insulin or control media, biopsied tissues were immunostained for Akt phosphorylation at Thr-308/9 (pAkt) and then imaged by confocal fluorescence microscopy automated to collect a large grid of high resolution fields. In hSAT from 40 men and women with obesity, substantial heterogeneity of pAkt densities in adipocyte membranes were quantified in each image mosaic using a spatial unit of at least twice the size of the point spread function. Statistical analysis of the distribution of pAkt spatial units was best fit as the weighted sum of two separate distributions, corresponding to either a low or high pAkt density. A "high pAkt fraction" metric was calculated from the fraction of high pAkt distributed units over the total units. Importantly, upon insulin stimulation, tissues from the same biopsy showed either a minimal or a substantial change in the high pAkt fraction. Further supporting a two-state response to insulin stimulation, subjects with similar insulin sensitivity indices are also segregated into either of two clusters identified by the amount of membrane-localized pAkt.

• Authors: Chad D McCormick, Hang N Waters, Ludmila Bezrukov, Roxana Taginya, Viraj Parikh, Ginikanwa I Onyekaba, Jordan A Levine, Andrew P Demidowich, Jack A Yanovski, Paul S Blank, Joshua Zimmerberg
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 293 Issue 37 Pg. 14249-14259 (09 14 2018) ISSN: 1083-351X [Electronic] United States
• PMID: 30006347 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, N.I.H., Intramural)
• Copyright: © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• Glucose Transporter Type 4
• Insulin
• SLC2A4 protein, human
• Proto-Oncogene Proteins c-akt

Topics

• Adipocytes (enzymology, metabolism)
• Adult
• Aged
• Cell Membrane (metabolism)
• Cohort Studies
• Enzyme Activation
• Female
• Glucose Transporter Type 4 (metabolism)
• Humans
• Insulin (metabolism)
• Insulin Resistance
• Male
• Microscopy, Confocal
• Microscopy, Fluorescence
• Middle Aged
• Obesity (metabolism)
• Phosphorylation
• Proto-Oncogene Proteins c-akt (metabolism)
• Signal Transduction
• Subcutaneous Fat (enzymology, metabolism)
• Young Adult