Manipulation and Measurement of AMPK Activity in Pancreatic Islets

Methods Mol Biol. 2018:1732:413-431. doi: 10.1007/978-1-4939-7598-3_26.

Abstract

The role of the energy sensor AMPK-activated protein kinase (AMPK) in the insulin-secreting β-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.

Keywords: AMPK activity; Insulin secretion; Islet isolation; Mouse and human islets; Pharmacological AMPK activators.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Aminoimidazole Carboxamide / analogs & derivatives
  • Aminoimidazole Carboxamide / pharmacology
  • Animals
  • Cell Line
  • Enzyme Activation / drug effects*
  • Enzyme Activators / pharmacology
  • Enzyme Assays / instrumentation
  • Enzyme Assays / methods*
  • Glucose / metabolism
  • Humans
  • Insulin / metabolism*
  • Insulin-Secreting Cells / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Primary Cell Culture
  • Ribonucleotides / pharmacology

Substances

  • Enzyme Activators
  • Insulin
  • Ribonucleotides
  • Aminoimidazole Carboxamide
  • AMP-Activated Protein Kinases
  • AICA ribonucleotide
  • Glucose