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European Journal of Biochemistry
Article . 2004 . Peer-reviewed
License: Wiley Online Library User Agreement
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Colocalization of insulin receptor and insulin receptor substrate‐1 to caveolae in primary human adipocytes

Cholesterol depletion blocks insulin signalling for metabolic and mitogenic control
Authors: Margareta, Karlsson; Hans, Thorn; Anna, Danielsson; Karin G, Stenkula; Anita, Ost; Johanna, Gustavsson; Fredrik H, Nystrom; +1 Authors

Colocalization of insulin receptor and insulin receptor substrate‐1 to caveolae in primary human adipocytes

Abstract

Caveolae are plasma membrane invaginations with several functions, one of which appears to be to organize receptor mediated signalling. Here we report that in primary human subcutaneous adipocytes the insulin receptor was localized to caveolae by electron microscopy/immunogold detection and by isolating caveolae from plasma membranes. Part of insulin receptor substrate 1 (IRS1), the immediate downstream signal mediator, was colocalized with the insulin receptor in the plasma membrane and caveolae, as demonstrated by immunofluorescence microscopy, immunogold electron microscopy, and immunogold electron microscopy of transfected recombinant HA‐IRS1. In contrast, rat epididymal adipocytes lacked IRS1 at the plasma membrane. Depletion of cholesterol from the cells using β‐cyclodextrin blocked insulin stimulation of glucose uptake, insulin inhibition of perilipin phosphorylation in response to isoproterenol, and insulin stimulation of protein kinase B and Map‐kinases extracellular signal‐related kinase (ERK)1/2 phosphorylation. Insulin‐stimulated phosphorylation of the insulin receptor and IRS1 was not affected, indicating that caveolae integrity is required downstream of IRS1. In conclusion we show that insulin receptor and IRS1 are both caveolar proteins and that caveolae are required for both metabolic and mitogenic control in human adipocytes. Our results establish caveolae as foci of insulin action and stress the importance of examining human cells in addition to animal cells and cell lines.

Related Organizations
Keywords

Adult, Male, Perilipin-1, Cell Membrane, Middle Aged, Caveolae, Phosphoproteins, Receptor, Insulin, Rats, Cholesterol, Adipose Tissue, Insulin Receptor Substrate Proteins, Animals, Humans, Insulin, Female, Phosphorylation, Carrier Proteins, Cells, Cultured, Aged

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    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    85
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
85
Top 10%
Top 10%
Top 10%
bronze