Investigative Ophthalmology & Visual Science, Vol 38, 1883-1892, Copyright © 1997 by Association for Research in Vision and Ophthalmology

ARTICLES AND REPORTS

Characterization of maxi-K-channels in bovine trabecular meshwork and their activation by cyclic guanosine monophosphate

F Stumpff, O Strauss, M Boxberger and M Wiederholt
Institut fur klinische Physiologie, Universitatsklinikum Benjamin Franklin, Freie Universitat Berlin, Germany.

PURPOSE: Electrophysiological characterization of trabecular meshwork cells and investigation of their response to elevation of cytosolic cyclic guanosine monophosphate (cGMP). METHODS: Bovine trabecular meshwork cells were cultured according to established methods and were studied, using the whole-cell and single-channel configurations of the patch-clamp technique. RESULTS: In single-channel experiments, cells expressed a channel with characteristics typical of maxi-K-channels. The channel was densely distributed in the membrane and had a high conductance of 326 +/- 4 pS (Pico Siemens) (symmetrical 150 mmol/l KCl; 37 degrees C) for potassium and negligible conductance for sodium (0.9 +/- 1 pS). The open probability could be elevated by depolarization, increasing cytosolic calcium, or adding adenosine triphosphate (1 mmol/ l). The channel could be blocked by external charybdotoxin (10(-8) mol/1), external TEA+ tetraethyl ammonium chloride (1 mmol/l) and by internal Ba2+ (10 mmol/l), whereas external Ba2+ and internal TEA+ (10 mmol/l) had no effect. In whole-cell experiments, trabecular meshwork cells displayed a strong outward conductance. Part of this conductance (35 +/- 5%) could be blocked by charybdotoxin and stimulated by ionomycin (10(-5) mol/1). Addition of 8-bromo-cGMP (10(-3) mol/1) stimulated the current to 290 +/- 57% (n = 4) of the original level, charybdotoxin led to a reduction of this current to 156 +/- 28% of the initial value. CONCLUSIONS: Trabecular meshwork cells express maxi-K- channels. These channels can be stimulated by raising internal cGMP levels and are known for their importance in smooth muscle relaxation. The results in this study supply further evidence that trabecular meshwork displays smooth muscle-like properties and contributes to the clarification of the mechanism leading to the relaxation of trabecular meshwork by nitrate and nonnitrate vasodilatators.

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