Reactive oxygen species produced by Zn2+ influx after exposure to AMPA, but not NMDA and their capturing effect on nigral dopaminergic protection

Abstract

Glutamate excitotoxicity is involved in dopaminergic degeneration in the substantia nigra pars compacta (SNpc). Here we compared vulnerability to neurodegeneration after exposure to NMDA and AMPA. Apomorphine-induced movement disorder and dopaminergic degeneration in the SNpc, which are associated with Parkinson's syndrome, were induced after injection of AMPA into the SNpc of rats, but not after injection of NMDA. Co-injection of 1-naphthyl acetyl spermine (NASPM), a selective blocker of Ca²⁺- and Zn²⁺-permeable GluR2-lacking AMPA receptors rescued dopaminergic degeneration and increase in intracellular Zn²⁺ by AMPA. Furthermore, we tested the effect of capturing reactive oxygen species (ROS) produced by Zn²⁺ on neuroprotection in vivo. The levels of ROS, which were determined by HYDROP, a membrane-permeable H₂O₂ fluorescence probe and Aminophenyl Fluorescein (APF), a fluorescence probe for hydroxyl radical and peroxynitrite, were increased after injection of AMPA, but not after co-injection of CaEDTA, an extracellular Zn²⁺ chelator, suggesting that increase in Zn²⁺ influx by AMPA elevates the levels of intracellular ROS. AMPA-mediated dopaminergic degeneration was completely rescued by co-injection of either HYDROP or APF. The present study indicates that neurotoxic signaling of the influx of extracellular Zn²⁺ through Zn²⁺-permeable GluR2-lacking AMPA receptors is converted to ROS production and that capturing the ROS completely protects dopaminergic degeneration after exposure to AMPA, but not NMDA. It is likely that regulation of the conversion from Zn²⁺ influx into ROS production plays a key role to preventing Parkinson's syndrome.

Keywords: AMPA receptor; NASPM; Nigral dopaminergic neuron; Parkinson's syndrome; Reactive oxygen species; Zn(2+) dysregulation.