Automated Patch Clamp Analysis of nAChα7 and Na(V)1.7 Channels

Curr Protoc Pharmacol. 2014 Jun 16:65:11.13.1-48. doi: 10.1002/0471141755.ph1113s65.

Abstract

Automated patch clamp devices are now commonly used for studying ion channels. A useful modification of this approach is the replacement of the glass pipet with a thin planar glass layer with a small hole in the middle. Planar patch clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high-quality and information-rich data, and allow for rapid perfusion and temperature control. Covered in this unit are two challenging targets in drug discovery: voltage-gated sodium subtype 1.7 (Na(V)1.7) and nicotinic acetylcholine α7 receptors (nAChα7R). Provided herein are protocols for recording activation and inactivation kinetics of Na(V)1.7, and activation and allosteric modulation of nAChα7R.

Keywords: NaV1.7; automated patch clamp; nicotinic acetylcholine receptors.

MeSH terms

  • Animals
  • Automation, Laboratory
  • CHO Cells / physiology
  • Cricetulus
  • HEK293 Cells / physiology
  • Humans
  • NAV1.7 Voltage-Gated Sodium Channel / physiology*
  • Patch-Clamp Techniques / methods*
  • Patch-Clamp Techniques / standards
  • alpha7 Nicotinic Acetylcholine Receptor / physiology*

Substances

  • NAV1.7 Voltage-Gated Sodium Channel
  • alpha7 Nicotinic Acetylcholine Receptor