Aim 1 will determine of AQP-1 is involved in the volume regulation of cells from the human TN and cells from the inner wall of the Schlemm s canal (IWSC) by affecting their permeability to water. This will be evaluated by electronic cell sizing following treatment with mercurials, anti-AQP-1 Fab fragments and/or antisense nucleotides to AQP-1. There are several weaknesses. No specific comments are made about the tonicity concentrations that will be used for the experiments. These are critical details which should be included in a table format assuming various tonicities will be attempted. Second, there is no mention about the specificity of the antibodies that will be utilized. Do they react to any of the other water channel proteins? Aim 2 will determine the contribution of AQP-1 to the bulk outflow of aqueous humor through the conventional outflow pathway. The anterior chamber will be analyzed in a perfusion chamber in which aqueous humor flows through the outflow pathway. Volume will be measured following treatment with mercurials, anti-AQP-1 Fab fragments and/or antisense nucleotides to AQP-1. These are valuable experiments and will be the most difficult. The candidate has reviewed the problems he might encounter especially with antisense oligonucleotides. Again, no mention is made of concentrations of antibodies, antisense nucleotides or mercurials is given. These will be important parameters that the candidate will have to address for the outcome to be successful. The scientific merit of the proposal is excellent.
