Morphological Identification of Melanopsin-Expressing Retinal Ganglion Cell Subtypes in Mice

Methods Mol Biol. 2018:1753:275-287. doi: 10.1007/978-1-4939-7720-8_19.

Abstract

Melanopsin-expressing, intrinsically photosensitive retinal ganglion cells (ipRGCs) are a relatively recently discovered class of photoreceptor. ipRGCs can be subdivided into at least five subtypes (M1-M5), each of which has a distinct complement of morphological and physiological properties. ipRGC subtypes can be identified morphologically based on a combination of dendritic morphology and immunostaining for a cell-type specific marker. In this chapter, we describe methods for conclusively identifying each of the five ipRGC subtypes through a combination of patch clamp electrophysiology, Neurobiotin filling, visualization of ipRGC dendrites, and immunostaining for the marker SMI-32.

Keywords: Dendritic stratification; Intrinsically photosensitive retinal ganglion cell; Melanopsin; Mouse; Opn4; Patch clamp electrophysiology; Retina; SMI-32; Subtypes.

MeSH terms

  • Animals
  • Biotin / analogs & derivatives
  • Biotin / chemistry
  • Dendrites / metabolism*
  • Mice
  • Mice, Transgenic
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods*
  • Patch-Clamp Techniques / instrumentation
  • Patch-Clamp Techniques / methods*
  • Retinal Ganglion Cells / metabolism*
  • Rod Opsins / genetics
  • Rod Opsins / metabolism*

Substances

  • Rod Opsins
  • melanopsin
  • neurobiotin
  • Biotin