Microglia Analysis in Retinal Degeneration Mouse Models

Methods Mol Biol. 2018:1753:159-166. doi: 10.1007/978-1-4939-7720-8_10.

Abstract

Microgliosis is a hallmark of degenerative processes in the retina. Reactive microglia migrate to the photoreceptor layer and the subretinal space during outer retinal degeneration. This process creates a toxic milieu where reactive microglia and dying photoreceptors recruit additional reactive phagocytes. This results in the release of a multitude of proinflammatory factors which accelerate photoreceptor demise. In this chapter, we outline in detail how to monitor microgliosis in the Fam161a-deficient mouse model of Retinitis Pigmentosa by performing immunohistochemical stainings of retinal cryosections and flat mounts using the marker Iba1. This protocol will serve as a guideline in evaluating microglia reactivity and localization in various mouse models of retinal degeneration.

Keywords: Cryosections; Fam161a; Flat mounts; Iba1; Immunohistochemistry; Microglia; Retinal degeneration.

MeSH terms

  • Animals
  • Calcium-Binding Proteins / analysis
  • Disease Models, Animal
  • Eye Proteins / genetics
  • Humans
  • Immunohistochemistry / instrumentation
  • Immunohistochemistry / methods*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microfilament Proteins / analysis
  • Microglia / pathology*
  • Photoreceptor Cells / pathology*
  • Retinitis Pigmentosa / genetics
  • Retinitis Pigmentosa / pathology*

Substances

  • Aif1 protein, mouse
  • Calcium-Binding Proteins
  • Eye Proteins
  • Fam161A protein, mouse
  • Microfilament Proteins