Inhibition of glutamate uptake causes an acute increase in aqueous humor protein.

Inhibition of glutamate transport has been shown to increase paracellular permeability of epithelial cell monolayers in vitro. To determine if blocking glutamate transport would affect tissue permeability in vivo, D-aspartate (D-Asp; 300 nmol 30 microliters-1) (a non-toxic competitive inhibitor of glutamate transport) or a placebo was injected into the anterior chambers of the fellow eyes of 15 adult rabbits. [14C]-L-glucose and/or [125I]-rabbit albumin were included in the injection vehicle as aqueous humor (AH) outflow markers. The specific inhibition of glutamate uptake by D-Asp was indicated by a 15% increase in AH glutamate (174 +/- 9 nmol ml-1 to 205 +/- 13 nmol ml-1; P = 0.03) at 1-1.5 hr post injection. Also, the efflux of [14C]-L-glucose and [125I]-rabbit albumin from the AH of D-Asp injected eyes was increased 22% over the placebo-injected control eyes (P < or = 0.02). Concomitantly, the total protein concentration in the AH from D-Asp injected eyes (517 +/- 35 micrograms ml-1) was 19% greater (P < 0.02) than the protein concentration in AH from placebo-injected control eyes (420 +/- 36 micrograms ml-1). In additional studies, an irreversible inhibitor of glutamate transport, threo-beta-hydroxyaspartate (THA; 30 nmol 30 microliters-1), was shown to increase the efflux of [14C]-L-glucose (22%; P < 0.05) from the anterior chamber and increase AH protein concentrations by 29% (484 +/- 112 micrograms ml-1 in control AH versus 686 +/- 117 micrograms ml-1 in THA AH, P = 0.08) at 1 hr post intracameral injection. SDS-PAGE analysis of the AH associated the protein increase in the D-Asp and THA injected eyes but not placebo-injected control eyes with a detectable increase in a 66 kDa protein (aligns with serum albumin) and several lower molecular weight (23-35 kDa) AH proteins. The results found suggest that inhibition of glutamate transport from the AH acutely increases intraocular epithelial/endothelial paracellular permeability.