Critical neuroprotective roles of heme oxygenase-1 induction against axonal injury-induced retinal ganglion cell death

J Neurosci Res. 2014 Sep;92(9):1134-42. doi: 10.1002/jnr.23398. Epub 2014 May 5.

Abstract

Although axonal damage induces significant retinal ganglion cell (RGC) death, small numbers of RGCs are able to survive up to 7 days after optic nerve crush (NC) injury. To develop new treatments, we set out to identify patterns of change in the gene expression of axonal damage-resistant RGCs. To compensate for the low density of RGCs in the retina, we performed retrograde labeling of these cells with 4Di-10ASP in adult mice and 7 days after NC purified the RGCs with fluorescence-activated cell sorting. Gene expression in the cells was determined with a microarray, and the expression of Ho-1 was determined with quantitative PCR (qPCR). Changes in protein expression were assessed with immunohistochemistry and immunoblotting. Additionally, the density of Fluoro-gold-labeled RGCs was counted in retinas from mice pretreated with CoPP, a potent HO-1 inducer. The microarray and qPCR analyses showed increased expression of Ho-1 in the post-NC RGCs. Immunohistochemistry also showed that HO-1-positive cells were present in the ganglion cell layer (GCL), and cell counting showed that the proportion of HO-1-positive cells in the GCL rose significantly after NC. Seven days after NC, the number of RGCs in the CoPP-treated mice was significantly higher than in the control mice. Combined pretreatment with SnPP, an HO-1 inhibitor, suppressed the neuroprotective effect of CoPP. These results reflect changes in HO-1 activity to RGCs that are a key part of RGC survival. Upregulation of HO-1 signaling may therefore be a novel therapeutic strategy for glaucoma.

Keywords: glaucoma; heme oxygenase-1; neuroprotection; retinal ganglion cell.

MeSH terms

  • Animals
  • Axons / pathology*
  • Disease Models, Animal
  • Flow Cytometry
  • Gene Expression Profiling
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / physiology*
  • Heme Oxygenase-1 / genetics
  • Heme Oxygenase-1 / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neuroprotective Agents / pharmacology
  • Neuroprotective Agents / therapeutic use
  • Oligonucleotide Array Sequence Analysis
  • Optic Nerve Injuries / drug therapy
  • Optic Nerve Injuries / pathology*
  • Protoporphyrins
  • Pyrazines / pharmacology
  • Pyrazines / therapeutic use
  • Pyrroles / pharmacology
  • Pyrroles / therapeutic use
  • Retinal Ganglion Cells / pathology*
  • Statistics, Nonparametric
  • Stilbamidines
  • Time Factors

Substances

  • 2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt
  • Nerve Tissue Proteins
  • Neuroprotective Agents
  • Protoporphyrins
  • Pyrazines
  • Pyrroles
  • Stilbamidines
  • cyclohexyl-octahydro-pyrrolo(1,2-a)pyrazine
  • cobaltiprotoporphyrin
  • Heme Oxygenase-1