Monitoring of rhodopsin trafficking and mistrafficking in live photoreceptors

Methods Mol Biol. 2015;1271:293-307. doi: 10.1007/978-1-4939-2330-4_19.

Abstract

Outer segment (OS) directed trafficking is required for accomplishing the extremely high concentration of rhodopsin and explicitly high photon sensitivity of rod photoreceptor cells. Aberrant targeting of rhodopsin often leads to blinding disorders, due to various mechanisms causing rhodopsin mislocalization. Until recently, it has been challenging to monitor the dynamics of rhodopsin biogenesis and trafficking. Here, we describe a new method to visualize rhodopsin trafficking in living and unfixed Xenopus laevis rod photoreceptors. By harnessing the photochemical property of a photoconvertible fluorescent protein Dendra2, it is now possible to encode temporal information into colors and resolve spatiotemporal distribution of rhodopsin-Dendra2 fusion proteins in individual rod photoreceptors.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Microscopy, Confocal
  • Protein Transport / physiology
  • Retinal Rod Photoreceptor Cells / metabolism*
  • Rhodopsin / chemistry
  • Rhodopsin / metabolism*
  • Xenopus laevis

Substances

  • Rhodopsin