Whole cell recording from an organotypic slice preparation of neocortex

Robert Foehring, Dongxu Guan, Tara Toleman, Angela R. Cantrell

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex. Because of the lack of specific pharmacological agents for these channels, we have taken a genetic approach to manipulating channel expression. We use an organotypic culture preparation (16) in order to maintain cell morphology and the laminar pattern of cortex. We typically isolate acute neocortical slices at postnatal days 8-10 and maintain the slices in culture for 3-7 days. This allows us to study neurons at a similar age to those in our work with acute slices and minimizes the development of exuberant excitatory connections in the slice. We record from visually-identified pyramidal neurons in layers II/III or V using infrared illumination (IR-) and differential interference contrast microscopy (DIC) with whole cell patch clamp in current- or voltage-clamp. We use biolistic (Gene gun) transfection of wild type or mutant potassium channel DNA to manipulate expression of the channels to study their function. The transfected cells are easily identified by epifluorescence microscopy after co-transfection with cDNA for green fluorescent protein (GFP). We compare recordings of transfected cells to adjacent, untransfected neurons in the same layer from the same slice.

Original languageEnglish (US)
Article numbere2600
JournalJournal of Visualized Experiments
Issue number52
DOIs
StatePublished - Jan 1 2011

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Neocortex
Patch-Clamp Techniques
Neurons
Pyramidal Cells
Clamping devices
Transfection
Potassium
Microscopic examination
Biolistics
Interference Microscopy
Voltage-Gated Potassium Channels
Potassium Channels
Firearms
Electric potential
Green Fluorescent Proteins
Lighting
Rats
Microscopy
DNA
Complementary DNA

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Whole cell recording from an organotypic slice preparation of neocortex. / Foehring, Robert; Guan, Dongxu; Toleman, Tara; Cantrell, Angela R.

In: Journal of Visualized Experiments, No. 52, e2600, 01.01.2011.

Research output: Contribution to journalArticle

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