Simultaneous Real-Time Measurement of the β-Cell Membrane Potential and CaInflux to Assess the Role of Potassium Channels on β-Cell Function.
AUTHORS
- PMID: 29058185 [PubMed].
ABSTRACT
Stimulus-secretion coupling in pancreatic β-cells requires Cainflux through voltage-dependent Cachannels, whose activity is controlled by the plasma membrane potential (V). Here, we present a method of measuring fluctuations in the β-cell Vand Cainflux simultaneously, which provides valuable information about the ionic signaling mechanisms that underlie insulin secretion. This chapter describes the use of perforated patch clamp electrophysiology on cells loaded with a fluorescent intracellular Caindicator, which permits the stable recording conditions needed to monitor the Vand Cainflux in β-cells. Moreover, this chapter describes the protocols necessary for the preparation of mouse and human islet cells for the simultaneous recording of Vand Caas well as determining the specific islet cell type assessed in each experiment.
Stimulus-secretion coupling in pancreatic β-cells requires Cainflux through voltage-dependent Cachannels, whose activity is controlled by the plasma membrane potential (V). Here, we present a method of measuring fluctuations in the β-cell Vand Cainflux simultaneously, which provides valuable information about the ionic signaling mechanisms that underlie insulin secretion. This chapter describes the use of perforated patch clamp electrophysiology on cells loaded with a fluorescent intracellular Caindicator, which permits the stable recording conditions needed to monitor the Vand Cainflux in β-cells. Moreover, this chapter describes the protocols necessary for the preparation of mouse and human islet cells for the simultaneous recording of Vand Caas well as determining the specific islet cell type assessed in each experiment.
Tags: 2018