Memantine blocks mitochondrial OPA1 and cytochrome c release and subsequent apoptotic cell death in glaucomatous retina

Invest Ophthalmol Vis Sci. 2009 Feb;50(2):707-16. doi: 10.1167/iovs.08-2499. Epub 2008 Oct 20.

Abstract

Purpose: To determine whether intraocular pressure (IOP) elevation alters OPA1 expression and triggers OPA1 release, as well as whether the uncompetitive N-methyl-d-aspartate (NMDA) glutamate receptor antagonist memantine blocks OPA1 release and subsequent apoptotic cell death in glaucomatous DBA/2J mouse retina.

Methods: Preglaucomatous DBA/2J mice received memantine (5 mg/kg, intraperitoneal injection, twice daily for 3 months) and IOP in the eyes was measured monthly. RGC loss was counted after FluoroGold labeling. OPA1, Dnm1, Bcl-2, and Bax mRNA were measured by qPCR. OPA1 protein was assessed by immunohistochemistry and Western blot. Apoptotic cell death was assessed by TUNEL staining.

Results: Memantine treatment significantly increased RGC survival in glaucomatous DBA/2J mice and increased the 75-kDa OPA1 isoform, but did not alter the 80- and 90-kDa isoforms. The isoforms of OPA1 were significantly increased in the cytosol of the vehicle-treated glaucomatous retinas but were significantly decreased in memantine-treated glaucomatous retinas. OPA1 immunoreactivity was decreased in the photoreceptors of both vehicle- and memantine-treated glaucomatous retinas, but was increased in the outer plexiform layer of only the memantine-treated glaucomatous retinas. Memantine blocked apoptotic cell death in the GCL, increased Bcl-2 gene expression, and decreased Bax gene expression.

Conclusions: OPA1 release from mitochondria in glaucomatous mouse retina is inhibited by blockade of glutamate receptor activation. Because this OPA1 effect was accompanied by increased Bcl-2 expression, decreased Bax expression, and apoptosis blockade, glutamate receptor activation in the glaucomatous retina may involve a distinct mitochondria-mediated cell death pathway.

Publication types

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

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Blotting, Western
  • Cell Survival
  • Cytochromes c / genetics
  • Cytochromes c / metabolism*
  • Dynamin I / genetics
  • Excitatory Amino Acid Antagonists / pharmacology
  • Female
  • GTP Phosphohydrolases / genetics
  • GTP Phosphohydrolases / metabolism*
  • Gene Expression
  • Glaucoma / genetics
  • Glaucoma / metabolism
  • Glaucoma / prevention & control*
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Injections, Intraperitoneal
  • Intraocular Pressure
  • Memantine / pharmacology*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred DBA
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • RNA, Messenger / metabolism
  • Receptors, N-Methyl-D-Aspartate / antagonists & inhibitors
  • Retinal Diseases / genetics
  • Retinal Diseases / metabolism
  • Retinal Diseases / prevention & control*
  • Retinal Ganglion Cells / drug effects
  • bcl-2-Associated X Protein / genetics

Substances

  • Bax protein, mouse
  • Excitatory Amino Acid Antagonists
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • Receptors, N-Methyl-D-Aspartate
  • bcl-2-Associated X Protein
  • Cytochromes c
  • Dynamin I
  • GTP Phosphohydrolases
  • Opa1 protein, mouse
  • Memantine