A guide to in vivo single-unit recording from optogenetically identified cortical inhibitory interneurons

J Vis Exp. 2014 Nov 7:(93):e51757. doi: 10.3791/51757.

Abstract

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral cortex. We here share our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex using a method developed by Lima and colleagues. Recordings are performed in mice expressing Channelrhodopsin-2 (ChR2) in specific neuronal subpopulations. Members of the population are identified by their response to a brief flash of blue light. This technique - termed "PINP", or Photostimulation-assisted Identification of Neuronal Populations - can be implemented with standard extracellular recording equipment. It can serve as an inexpensive and accessible alternative to calcium imaging or visually-guided patching, for the purpose of targeting extracellular recordings to genetically-identified cells. Here we provide a set of guidelines for optimizing the method in everyday practice. We refined our strategy specifically for targeting parvalbumin-positive (PV+) cells, but have found that it works for other interneuron types as well, such as somatostatin-expressing (SOM+) and calretinin-expressing (CR+) interneurons.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Calbindin 2 / biosynthesis
  • Cerebral Cortex / cytology
  • Cerebral Cortex / metabolism
  • Cerebral Cortex / physiology*
  • Channelrhodopsins
  • Fiber Optic Technology / methods*
  • Interneurons / cytology
  • Interneurons / metabolism
  • Interneurons / physiology*
  • Membrane Proteins / biosynthesis
  • Mice
  • Mice, Transgenic
  • Neurophysiology / methods*
  • Phosphoproteins / biosynthesis
  • Somatostatin / biosynthesis

Substances

  • Calb2 protein, mouse
  • Calbindin 2
  • Channelrhodopsins
  • Membrane Proteins
  • Palm protein, mouse
  • Phosphoproteins
  • Somatostatin