Concentration-Dependent Inner Retina Layer Damage and Optic Nerve Degeneration in a NMDA Model.

The intravitreal injection of N-methyl-D-aspartate (NMDA), a glutamate analogue, is an established model for fast retinal ganglion cell (RGC) degeneration. Yet, NMDA does not cause specific RGC damage. Now, the effects on the whole retina were analyzed. Additionally, the related effects for the structure and apoptotic levels of the optic nerve were investigated. Therefore, different NMDA concentrations were intravitreally injected in rats (20, 40, or 80 nmol NMDA or PBS). At days 3 and 14, Brn-3a+ RGCs were degenerated. A damage of calretinin+ amacrine cells was also recognized at day 14. Only a slight damage was observed in regard to PKCα+ bipolar cells, while rhodopsin+ photoreceptors remained intact. A long-lasting retinal microglia response was observed from day 3 up to day 14. Furthermore, a partial degeneration of the optic nerve was noted. At day 3, the SMI-32+ neurofilaments were just slightly affected, whereas the neurofilament structure was further degenerated at day 14. However, the luxol fast blue (LFB)-stained myelin structure remained intact from day 3 up to day 14. Interestingly, apoptotic mechanisms, like FasL and Fas co-localization as well as caspase 3 activation, were restricted to the optic nerve of the highest NMDA group at this late stage of degeneration. The degeneration of the optic nerve is probably only a side effect of neuronal degeneration of the inner retinal layers. The intact myelin structure might form a barrier against the direct influence of NMDA. In conclusion, this model is very suitable to test therapeutic agents, but it is important to analyze all inner retina layers and the optic nerve to determine their efficacy in this model more precisely.