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Effect of Ocular Hypertension on Retinal Nitridergic Pathway Activity

¹From the Laboratorio de Neuroquímica Retiniana y Oftalmología Experimental, the ²Departamento de Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, CEFyBO (Centro de Estudios Farmacológicos y Botánicos), CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), Buenos Aires, Argentina.

PURPOSE. Understanding the mechanisms of neuronal cell death in glaucoma is important for devising new treatments. Excitatory amino acids, excessive Ca²⁺ influx, and formation of nitric oxide (NO) via NO synthase (NOS)-1 could be involved in glaucomatous neuropathy. The purpose of the present study was to examine the retinal nitridergic pathway activity in rats exposed to experimentally elevated intraocular pressure.

METHODS. Weekly injections of HA were performed unilaterally in the rat anterior chamber, whereas the contralateral eye was injected with saline solution. At 3 or 6 weeks of treatment, retinal NOS activity was assessed through the conversion of ³H-L-arginine to ³H-L-citrulline, whereas NOS-1, -2, and -3 levels were assessed by Western blotting. L-Arginine uptake was measured using ³H-L-arginine, whereas mRNA levels of L-arginine transporters were determined by semiquantitative RT-PCR. In addition, cyclic guanosine monophosphate (cGMP) levels were quantified by radioimmunoassay.

RESULTS. At both 3 and 6 weeks of treatment, NOS activity significantly increased in HA-injected eyes although no changes in retinal NOS-1, -2, or -3 levels were observed in eyes injected with HA. L-Arginine influx and mRNA levels of cationic amino acid transporter type (CAT)-1 and -2 significantly increased in retinas from hypertensive eyes. Retinal cGMP levels significantly increased in eyes injected with HA for 3 but not 6 weeks.

CONCLUSIONS. These results suggest a significant activation of the retinal nitridergic pathway in hypertensive eyes.

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