dizocilpine-maleate and Retinal-Degeneration

Apolipoprotein E (ApoE) plays important roles in the body, including a carrier of cholesterols, an anti-oxidant, and a ligand for the low-density lipoprotein receptors. In the nervous system, the presence of ApoE4 isoforms is associated with Alzheimer's disease. ApoE gene polymorphisms are also associated with glaucoma, but the function of ApoE in the retina remains unclear. In this study, we investigated the role of ApoE in axonal damage-induced RGC death. ApoE was detected in the astrocytes and Müller cells in the wild-type (WT) retina. RGC damage was induced in adult ApoE-deficient mice (male, 10-12 weeks old) through ocular hypertension (OH), optic nerve crush (NC), or by administering kainic acid (KA) intravitreally. The WT mice were treated with a glutamate receptor antagonist (MK801 or CNQX) 30 min before performing NC or left untreated. Seven days later, the retinas were flat mounted and Fluorogold-labeled RGCs were counted. We found that the RGCs in the ApoE-deficient mice were resistant to OH-induced RGC death and optic nerve degeneration 4 weeks after induction. In WT mice, NC effectively induced RGC death (control: 4085±331 cells/mm(2), NC: 1728±170 cells/mm(2)). CNQX, an inhibitor of KA receptors, suppressed this RGC death (3031±246 cells/mm(2)), but MK801, an inhibitor of NMDA receptors, did not (1769±212 cells/mm(2)). This indicated the involvement of KA receptor signaling in NC-induced RGC death. We found that NC- or KA-induced RGC death was significantly less in the ApoE-deficient mice than in the WT mice. These data suggest that the ApoE deficiency had a neuroprotective effect against axonal damage-induced RGC death by suppressing the KA receptor signaling.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Apolipoproteins E; Cell Death; Disease Models, Animal; Dizocilpine Maleate; Dose-Response Relationship, Drug; Kainic Acid; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Neuroprotective Agents; Ocular Hypertension; Optic Nerve Injuries; Receptors, Kainic Acid; Retinal Degeneration; Retinal Ganglion Cells; Signal Transduction; Stilbamidines

Previously we showed that apoptosis is involved in N -methyl- D -aspartate (NMDA) induced excitotoxicity in adult rat retinas. Since rabbits have a higher endogenous level of glutamate in the retina and very different retinal structures, it is not clear if apoptosis is similarly involved in adult rabbit retinas after intravitreal injection of NMDA. In this study, we used ultrastructural features, TdT-mediated biotin-dUTP nick end labeling (TUNEL) and two caspase inhibitors to examine whether apoptosis is involved in NMDA-induced excitotoxicity in adult rabbit retinas. At 18 hr after an intravitreal injection of 400 nmoles NMDA, typical apoptotic features in degenerative cells in the retinal ganglion cell layer (RGCL) and the inner nuclear layer (INL) were noted by electron microscopy. TUNEL positive nuclei were detected in these layers as early as 4 hr showing maximal numbers at 18 hr. At 7 days, significant loss of nuclei from the RGCL was noted at the visual streak, the superior and the inferior retinas. These losses were abolished by simultaneous administration of MK-801 and ameliorated by YVAD, a caspase-1 inhibitor, but not by IETD, a caspase-8 inhibitor. These results indicated that, similar to adult rat retinas, apoptosis is involved in NMDA receptor-mediated excitotoxicity in rabbit retinas and that specific caspases may play important roles.

Topics: Animals; Apoptosis; Caspase Inhibitors; Caspases; Dizocilpine Maleate; In Situ Nick-End Labeling; Microscopy, Electron; N-Methylaspartate; Neuroprotective Agents; Rabbits; Retina; Retinal Degeneration

To establish a new behavioral animal model of excitotoxicity, we injected adult rats intraocularly with a single dose of 2, 20, or 100 nmol of N-methyl-D-aspartate (NMDA). We quantified visual impairment by plotting the size of the visual field in which the rats successfully oriented towards a small, moving target. In comparison to the saline-injected (contralateral) control side, the side injected with 2 nmol of NMDA was not significantly impaired. When injected with higher doses, the rats were nearly blind immediately after surgery, with only about 20% (20 nmol NMDA) or 10% (100 nmol NMDA) of residual vision. Within about 3 weeks, however, visual performance returned to near-normal levels. Simultaneous intraocular administration of a non-competitive NMDA-antagonist, MK-801 (1 nmol), resulted in complete behavioral protection. NMDA administration led to a dose-dependent loss of cells within the ganglion cell layer, as assessed in whole-mounted retinae which were retrogradely labelled with horseradish peroxidase (HRP). Whereas 2 nmol of NMDA led to the loss of about 30% of retinal ganglion cells (RGCs), at higher NMDA doses only 13% of the RGCs survived. After the injection of 20 nmol of NMDA, large-diameter RGCs (> 22 microns) survived the lesion to a greater extent than small diameter cells (8-21 microns); at 100 nmol cells of all diameters were equally affected. The number of Nissl-stained cells with small diameters (< 11 microns), presumed to be displaced amacrine cells, was also affected by NMDA, although to a lesser degree. Analysis of behavioral performance (vision score) and the number of cells in the retina revealed a correlation of r = 0.76 between visual performance and the number of HRP-filled RGCs immediately after surgery. Lower correlations were found between visual performance and cells stained with Nissl of diameters smaller than 11 microns (presumed RGCs without retinofugal connections; r = 0.55 and r = 0.58, respectively). Because of the spontaneous recovery of vision, all correlations declined to values near 0 after 3 weeks. Thus, despite a dramatic loss of RGCs following NMDA administration, visual deficits recover significantly in adult rats within 2-3 weeks.

Topics: Animals; Behavior, Animal; Cell Survival; Dizocilpine Maleate; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Eye; Horseradish Peroxidase; Injections; N-Methylaspartate; Orientation; Rats; Rats, Inbred Strains; Retinal Degeneration; Retinal Ganglion Cells; Vision Disorders; Visual Fields

Pineal gland N-acetyltransferase (NAT) activity and pineal and serum levels of melatonin declined linearly in albino rats acutely exposed to different intensities of red light (600 nm or higher; low, 140 microW/cm2; moderate, 690 microW/cm2; high, 1200 microW/cm2) during the middle of the night. The high intensity red light was as effective as white light (780 microW/cm2) in suppressing NAT activity and pineal and circulating melatonin. Red-light-inhibited nighttime NAT activity and suppressed nocturnal melatonin levels in both retinally degenerate and normal rats. Pretreatment with the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (10 mg/kg intraperitoneally) completely prevented the red-light-induced inhibition of nighttime melatonin synthesis. Magnesium chloride (300 mg/kg intraperitoneally) reduced the inhibitory effects of low and moderate intensities of red light but was ineffective when high red-light intensity was used. However, both agents failed to antagonize the suppression of nighttime melatonin synthesis elicted by the exposure to white light. Since retinally degenerate and retinally normal animals respond in the same way to both red-light and pharmacological intervention with the NMDA receptor blocker MK-801, the findings indicate that the activation of central hypothalamic NMDA receptors might mediate the photic inhibition of nocturnal melatonin synthesis in the pineal gland elicited by the exposure to red light at night. Red-light-induced suppression of nocturnal melatonin synthesis possibly can be used to investigate the biochemical mechanisms by which light entrains melatonin synthesis and to study the pharmacological and physiological effects of endogenous and synthetic agents that antagonize the NMDA receptor response.

Topics: Animals; Arylamine N-Acetyltransferase; Dizocilpine Maleate; Dose-Response Relationship, Radiation; Excitatory Amino Acid Antagonists; Male; Melatonin; Neuroprotective Agents; Photic Stimulation; Pineal Gland; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Reference Values; Retina; Retinal Degeneration