Title

RNAi-based gene silencing in primary mouse and human adipose tissues

UMMS Affiliation

Program in Molecular Medicine; Department of Surgery

Date

11-10-2006

Document Type

Article

Subjects

Adaptor Proteins, Signal Transducing; Adipose Tissue, White; Animals; Cells, Cultured; Gene Expression Regulation; Glucose; Glucose Transporter Type 1; Glucose Transporter Type 4; Humans; Male; Mice; Nuclear Proteins; PTEN Phosphohydrolase; Phosphorylation; Proto-Oncogene Proteins c-akt; *RNA Interference; RNA, Messenger; Up-Regulation

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Cultured adipocyte cell lines are a model system widely used to study adipose function, but they exhibit significant physiological differences compared with primary cells from adipose tissue. Here we report short interfering RNA-based methodology to selectively attenuate gene expression in mouse and human primary adipose tissues as a means of rapidly validating findings made in cultured adipocyte cell lines. The method is exemplified by depletion of the PTEN phosphatase in white adipose tissue (WAT) from mouse and humans, which increases Akt phosphorylation as expected. This technology is also shown to silence genes in mouse brown adipose tissue. Previous work revealed upregulation of the mitochondrial protein UCP1 in adipose cells from mice lacking the gene for the transcriptional corepressor RIP140, whereas in cultured adipocytes, loss of RIP140 has a little effect on UCP1 expression. Application of our method to deplete RIP140 in primary mouse WAT elicited markedly increased oxygen consumption and expression of UCP1 that exactly mimics the phenotype observed in RIP140-null mice. This ex-vivo method of gene silencing should be useful in rapid validation studies as well as in addressing the depot- and species-specific functions of genes in adipose biology.

Rights and Permissions

Citation: J Lipid Res. 2007 Feb;48(2):465-71. Epub 2006 Nov 8. Link to article on publisher's site

DOI of Published Version

10.1194/jlr.D600033-JLR200

Related Resources

Link to Article in PubMed

Journal Title

Journal of lipid research

PubMed ID

17093294