Patch Clamp Protocols to Study Ion Channel Activity in Microglia
Tom Schilling, Claudia Eder
Abstract
Tom Schilling, Claudia Eder
Abstract
Microglia express a variety of ion channels, which can be distinguished based on their ion selectivity into K(+), H(+), Na(+), Ca(2+), nonselective cation, and Cl(-) channels. With respect to their activation mode, voltage-, Ca(2+)-, calcium release-, G protein-, swelling-, and stretch-activated ion channels have been described in microglia. The best method to study the activity of microglial ion channels is the patch clamp technique. The activity of microglial ion channels under physiological conditions is best explored using the perforated patch clamp technique, which allows recordings of membrane potential or ion currents, while the intracellular milieu of the cells remains intact. In whole-cell patch clamp recordings, application of specific voltage protocols with defined intra- and extracellular solutions allows precise identification of a certain ion channel type in microglia as well as the investigation of the channel's biophysical and pharmacological properties. This chapter summarizes patch clamp protocols optimal for recording and analysis of microglial ion channel activity in vitro and in situ.
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Microglia express a variety of ion channels, which can be distinguished based on their ion selectivity into K(+), H(+), Na(+), Ca(2+), nonselective cation, and Cl(-) channels. With respect to their activation mode, voltage-, Ca(2+)-, calcium release-, G protein-, swelling-, and stretch-activated ion channels have been described in microglia. The best method to study the activity of microglial ion channels is the patch clamp technique. The activity of microglial ion channels under physiological conditions is best explored using the perforated patch clamp technique, which allows recordings of membrane potential or ion currents, while the intracellular milieu of the cells remains intact. In whole-cell patch clamp recordings, application of specific voltage protocols with defined intra- and extracellular solutions allows precise identification of a certain ion channel type in microglia as well as the investigation of the channel's biophysical and pharmacological properties. This chapter summarizes patch clamp protocols optimal for recording and analysis of microglial ion channel activity in vitro and in situ.
Key concepts: Ion channel, Patch clamp, Microglia, Biophysics, Chemistry, Electrophysiology, Voltage clamp, Current clamp