Optogenetic Control of Pancreatic Islets

Methods Mol Biol. 2016:1408:107-23. doi: 10.1007/978-1-4939-3512-3_8.

Abstract

In light of the emerging diabetes epidemic, new experimental approaches in islet research are needed to elucidate the mechanisms behind pancreatic islet dysfunction and to facilitate the development of more effective therapies. Optogenetics has created numerous new experimental tools enabling us to gain insights into processes little was known about before. The spatial and temporal precision that it can achieve is also attractive for studying the cells of the pancreatic islet and we set out to explore the possibilities of this technology for our purposes. We here describe how to use the islets of an "optogenetic beta-cell" mouse line in islet batch incubations and Ca(2+) imaging experiments. This protocol enables light-induced insulin release and provides an all-optical solution to control and measure intracellular Ca(2+) levels in pancreatic beta-cells. The technique is easy to set up and provides a useful tool for controlling the activity of distinct islet cell populations.

Keywords: All-optical; Beta-cell; Calcium; Channelrhodopsin-2; Diabetes; Insulin; Islets; Optogenetics; Pancreas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / analysis*
  • Calcium / metabolism
  • Genotype
  • Insulin / metabolism
  • Insulin-Secreting Cells / cytology
  • Insulin-Secreting Cells / metabolism
  • Islets of Langerhans / cytology
  • Islets of Langerhans / metabolism*
  • Light
  • Mice
  • Mice, Transgenic
  • Optical Imaging / methods*
  • Optogenetics / methods*
  • Tissue Culture Techniques / methods

Substances

  • Insulin
  • Calcium