Measuring dendritic distribution of membrane proteins

Edmund W. Ballou*, W. Bryan Smith, Roberta Anelli, Cj Heckman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

Neurons perform much of their integrative work in the dendritic tree, and spinal motoneurons have the largest tree of any cell. Electrical excitability is strongly influenced by dendrite membrane properties, which are difficult to measure directly. We describe a method to measure the distribution of ion channel membrane densities along dendritic trajectories. The method combines standard immunohistochemistry with reconstruction procedures for both large-scale and small-scale optical microscopy. Software written for Matlab then extracts the colocalization of the target ion channel with the target dye injected cell, and calculates the relative channel density per square micron of cell surface area, as a function of distance from the cell body. The technique can be used to quantify the localization and distribution of any immunoreactive moiety, and the software provides a flexible vehicle for sensitivity analysis, to validate heuristics for selecting thresholds.

Original languageEnglish (US)
Pages (from-to)257-266
Number of pages10
JournalJournal of Neuroscience Methods
Volume156
Issue number1-2
DOIs
StatePublished - Sep 30 2006

Keywords

  • Colocalization
  • Dendrites
  • Distribution
  • Excitability
  • Ion channels
  • Membrane
  • Modeling
  • Motoneuron
  • Neuroanatomy
  • Neuron
  • Reconstruction

ASJC Scopus subject areas

  • Neuroscience(all)

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