Inhibition of N-Methyl-D-aspartate-induced Retinal Neuronal Death by Polyarginine Peptides Is Linked to the Attenuation of Stress-induced Hyperpolarization of the Inner Mitochondrial Membrane Potential

J Biol Chem. 2015 Sep 4;290(36):22030-48. doi: 10.1074/jbc.M115.662791. Epub 2015 Jun 22.

Abstract

It is widely accepted that overactivation of NMDA receptors, resulting in calcium overload and consequent mitochondrial dysfunction in retinal ganglion neurons, plays a significant role in promoting neurodegenerative disorders such as glaucoma. Calcium has been shown to initiate a transient hyperpolarization of the mitochondrial membrane potential triggering a burst of reactive oxygen species leading to apoptosis. Strategies that enhance cell survival signaling pathways aimed at preventing this adverse hyperpolarization of the mitochondrial membrane potential may provide a novel therapeutic intervention in retinal disease. In the retina, brain-derived neurotrophic factor has been shown to be neuroprotective, and our group previously reported a PSD-95/PDZ-binding cyclic peptide (CN2097) that augments brain-derived neurotrophic factor-induced pro-survival signaling. Here, we examined the neuroprotective properties of CN2097 using an established retinal in vivo NMDA toxicity model. CN2097 completely attenuated NMDA-induced caspase 3-dependent and -independent cell death and PARP-1 activation pathways, blocked necrosis, and fully prevented the loss of long term ganglion cell viability. Although neuroprotection was partially dependent upon CN2097 binding to the PDZ domain of PSD-95, our results show that the polyarginine-rich transport moiety C-R(7), linked to the PDZ-PSD-95-binding cyclic peptide, was sufficient to mediate short and long term protection via a mitochondrial targeting mechanism. C-R(7) localized to mitochondria and was found to reduce mitochondrial respiration, mitochondrial membrane hyperpolarization, and the generation of reactive oxygen species, promoting survival of retinal neurons.

Keywords: N-methyl-d-aspartate receptor (NMDA receptor, NMDAR); apoptosis; mitochondrial membrane potential; necrosis (necrotic death); neuroprotection; retina; retinal ganglion cells.

Publication types

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

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Death / drug effects
  • Disks Large Homolog 4 Protein
  • Excitatory Amino Acid Agonists / pharmacology
  • Guanylate Kinases / metabolism
  • HEK293 Cells
  • Humans
  • Male
  • Membrane Potential, Mitochondrial / drug effects*
  • Membrane Proteins / metabolism
  • Microscopy, Fluorescence
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Mitochondria / physiology
  • Mitochondrial Membranes / drug effects
  • Mitochondrial Membranes / physiology
  • N-Methylaspartate / pharmacology*
  • Neuroprotective Agents / metabolism
  • Neuroprotective Agents / pharmacology
  • Peptides / metabolism
  • Peptides / pharmacology*
  • Peptides, Cyclic / metabolism
  • Peptides, Cyclic / pharmacology
  • Protein Binding
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species / metabolism
  • Receptors, N-Methyl-D-Aspartate / metabolism
  • Retina / cytology
  • Retina / drug effects
  • Retina / metabolism
  • Retinal Ganglion Cells / drug effects
  • Retinal Ganglion Cells / metabolism
  • Retinal Neurons / drug effects*
  • Retinal Neurons / metabolism
  • Stress, Physiological / drug effects
  • Stress, Physiological / physiology

Substances

  • CN2097
  • Disks Large Homolog 4 Protein
  • Dlg4 protein, mouse
  • Excitatory Amino Acid Agonists
  • Membrane Proteins
  • Neuroprotective Agents
  • Peptides
  • Peptides, Cyclic
  • Reactive Oxygen Species
  • Receptors, N-Methyl-D-Aspartate
  • polyarginine
  • N-Methylaspartate
  • Guanylate Kinases