Native System and Cultured Cell Electrophysiology for Investigating Anesthetic Mechanisms

Methods Enzymol. 2018:602:301-338. doi: 10.1016/bs.mie.2018.01.012. Epub 2018 Mar 20.

Abstract

Anesthetic agents interact with a variety of ion channels and membrane-bound receptors, often at agent-specific binding sites of a single protein. These molecular-level interactions are ultimately responsible for producing the clinically anesthetized state. Between these two scales of effect, anesthetic agents can be studied in terms of how they impact the physiology of neuronal circuits, individual neurons, and cells expressing individual receptor types. The acutely dissected hippocampal slice is one of the most extensively studied and characterized preparations of intact neural tissue and serves as a highly useful experimental model system to test hypotheses of anesthetic mechanisms. Specific agent-receptor interactions and their effect on excitable membranes can further be defined with molecular precision in cell-based expression systems. We highlight several approaches in these respective systems that we have used and that also have been used by many investigators worldwide. We emphasize economy and quality control, to allow an experimenter to carry out these types of studies in a rigorous and efficient manner.

Keywords: Cultured cells; Dissociated neuron; Electrophysiology; Field recording; GABA(A) receptor; HEK293 cell; Hippocampus; Patch clamp; Pop spike; Transfection.

Publication types

  • Review

MeSH terms

  • Anesthetics / pharmacology*
  • Animals
  • Electrodes
  • Electrophysiology / economics
  • Electrophysiology / instrumentation
  • Electrophysiology / methods*
  • GABA-A Receptor Antagonists / pharmacology
  • HEK293 Cells
  • Hippocampus / cytology
  • Hippocampus / drug effects*
  • Hippocampus / metabolism
  • Histocytological Preparation Techniques / instrumentation
  • Histocytological Preparation Techniques / methods*
  • Humans
  • Mice
  • Models, Animal
  • Neurons / drug effects
  • Neurons / metabolism
  • Patch-Clamp Techniques / economics
  • Patch-Clamp Techniques / instrumentation
  • Patch-Clamp Techniques / methods
  • Primary Cell Culture / instrumentation
  • Primary Cell Culture / methods
  • Rats
  • Receptors, GABA-A / metabolism
  • Synaptic Potentials / drug effects

Substances

  • Anesthetics
  • GABA-A Receptor Antagonists
  • Receptors, GABA-A