Immunohistochemical and calcium imaging methods in wholemount rat retina

J Vis Exp. 2014 Oct 13:(92):e51396. doi: 10.3791/51396.

Abstract

In this paper we describe the tools, reagents, and the practical steps that are needed for: 1) successful preparation of wholemount retinas for immunohistochemistry and, 2) calcium imaging for the study of voltage gated calcium channel (VGCC) mediated calcium signaling in retinal ganglion cells. The calcium imaging method we describe circumvents issues concerning non-specific loading of displaced amacrine cells in the ganglion cell layer.

Publication types

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

MeSH terms

  • Animals
  • Axons / metabolism
  • Calcium / analysis*
  • Calcium / metabolism
  • Calcium Channels / metabolism
  • Calcium Signaling
  • Female
  • Immunohistochemistry / methods
  • Male
  • Rats
  • Rats, Sprague-Dawley
  • Retina / chemistry*
  • Retina / cytology
  • Retina / metabolism
  • Retinal Ganglion Cells / metabolism

Substances

  • Calcium Channels
  • Calcium