Biology Reference
In-Depth Information
Chapter 21
Application of DIGE and Mass Spectrometry in the Study
of Type 2 Diabetes Mellitus Mouse Models
Celia Smith , Davinia Mills , and Rainer Cramer
Abstract
Knowledge of the differences between the amounts and types of protein that are expressed in diseased
compared to healthy subjects may give an understanding of the biological pathways that cause disease.
This is the reasoning behind the presented protocol, which uses difference gel electrophoresis (DIGE)
to discover up- or down-regulated proteins between mice of different genotypes, or of those fed on
different diets, that may thus be prone to develop diabetes-like phenotypes. Subsequent analysis of
these proteins by tandem mass spectrometry typically facilitates their identifi cation with a high degree
of confi dence.
Key words:
Difference gel electrophoresis, Mass spectrometry, Proteomics, Quantitation, Type 2
diabetes mellitus
1. Introduction
Type 2 diabetes mellitus (T2DM) has reached epidemic proportions
globally, and the number of sufferers is expected to more than
double in the period 2000-2030 to 366 million worldwide(
1, 2
).
The protocol presented here combines difference gel electropho-
resis (DIGE) separation and quantitation with mass spectrometric
identifi cation of the proteins that are differentially expressed in lean
and obese mice when fed standard and high-fat diets. It is hoped
that the identifi cation of these proteins will help to unravel the
complex biological pathways involved in glycaemic control and
obesity, which are closely related to T2DM.
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