Dissociation of retinal ganglion cells without enzymes

Yuki Hayashida, Gloria J. Partida, Andrew Ishida

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

We describe here methods for dissociating retinal ganglion cells from adult goldfish and rat without proteolytic enzymes, and show responses of ganglion cells isolated this way to step-wise voltage changes and fluctuating current injections. Taking advantage of the laminar organization of vertebrate retinas, photoreceptors and other cells were lifted away from the distal side of freshly isolated goldfish retinas, after contact with pieces of membrane filter. Likewise, cells were sliced away from the distal side of freshly isolated rat retinas, after these adhered to a membrane filter. The remaining portions of retina were incubated in an enzyme-free, low Ca2+ solution, and triturated. After aliquots of the resulting cell suspension were plated, ganglion cells could be identified by dye retrogradely transported via the optic nerve. These cells showed no obvious morphological degeneration for several days of culture. Perforated-patch whole-cell recordings showed that the goldfish ganglion cells spike tonically in response to depolarizing constant current injections, that these spikes are temporally precise in response to fluctuating current injections, and that the largest voltage-gated Na+ currents of these cells were larger than those of ganglion cells isolated with a neutral protease.

Original languageEnglish (US)
Pages (from-to)25-35
Number of pages11
JournalJournal of Neuroscience Methods
Volume137
Issue number1
DOIs
StatePublished - Aug 15 2004

Keywords

  • Enzyme-free dissociation
  • Goldfish
  • Na current
  • Rat
  • Retinal ganglion cell
  • Spikes

ASJC Scopus subject areas

  • Neuroscience(all)

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