AMPK Methods and Protocols

Chapter 26

Manipulation and Measurement of AMPK Activity

in Pancreatic Islets

Aida Martinez-Sanchez, Marie-Sophie Nguyen-Tu, Isabelle Leclerc,

and Guy A. Rutter

Abstract

The role of the energy sensor AMPK-activated protein kinase (AMPK) in the insulin-secretingβ-cell
remains unclear and a subject of intense research. With this chapter, we aim to provide a detailed description
of the methods that our group routinely applies to the study of AMPK function in mouse and human
pancreatic islets. Thus, we provide detailed protocols to isolate and/or culture mouse and human islets, to
modulate and measure AMPK activity in isolated islets, and to evaluate its impact on islet function.

Key wordsMouse and human islets, Islet isolation, AMPK activity, Insulin secretion, Pharmacologi-

cal AMPK activators

1 Introduction

Pancreaticβ-cells are responsible for insulin release in response to

elevated glucose concentration in the blood and are therefore

essential for normal glucose homeostasis. Diabetes affects 8.5% of

the global population [1], and type 2 diabetes (T2D), fueled by the

global obesity epidemic, is one of the major modern health con-

cerns. Central to the development of diabetes is the failure of

β-cells, unable to secrete enough insulin to lower blood sugar

levels. A high prevailing blood sugar will itself further accelerate

β-cell failure in a vicious circle.

AMP-activated protein kinase (AMPK) is an energy sensor

involved in the control of glucose homeostasis and a suggested

target for some of the most commonly used antidiabetic drugs,

such as metformin, rosiglitazone, and the SGLT2 inhibitor canagli-

flozin [2, 3]. Despite the glucose-lowering effects of AMPK activa-

tion in the liver and muscle, the impact in other tissues is still

debated. In the β-cell, AMPK activity is promoted by glucose

deprivation [4], but its role in processes central toβ-cell function,

Dietbert Neumann and Benoit Viollet (eds.),AMPK: Methods and Protocols, Methods in Molecular Biology, vol. 1732,
https://doi.org/10.1007/978-1-4939-7598-3_26,©Springer Science+Business Media, LLC 2018

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