Experimental Study of Aqueous Outflow Facility : Eye Perfusion Technique and Tissue Culture Methods

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Outflow aqueous humor has been considered to pass through the trabecular meshwork(TM)down a pressure gradient without requiring pumping energy from the cells of the TM. However, trabecular outflow pathways may constitute more than merely a passive, structural filter for egress of aqueous humor from the eye.TM cells(2nd or 3rd subculture) were placed cultured in sparse for six days and then exposed to media tested in both the presence or absence of either catalase and superoxide dismutase etc. The effects were examined by changes of outflow facility and cell shape. In a parallel experiment, cell shape change was observed with various cytoskeleton affecting drugs and by cell cytoskeleton (actin) and by gel electrophoresis.To further understand the aqueous outflow mechanism, we also compared changes in the outflow facility with the tone and nerve-mediated contraction of ciliary muscle. NaF reduced the facility as much as 50%. lodoacetamide and iodoacetate both increased facility at 35 C. 2, 4 dinitrophenol, KCN and 2-deoxyglucose did not produce any effect. Theophylline and isobutylmethylxantnine increased outflow facility. However, neither epinephrine nor isoproterenol increased the, outflow facility, regardless of their concentrations. Epinephrine or isoproterenol(10^<-8>-10^<-6> M)did not relax the tone of bovine ciliary muscle, whereas theopliylline and IBMX remarkably relaxed the muscle which had been contracted by carbachol and also inhibited the nerve-mediated contraction.These findings suggest that several substances in the aqueous humor may influence outflow facility. Morphological change(cell shape or cell attachment)of outflow tissues and the induced response of ciliary muscle may play a significant role on the outflow facility.