Effects of Na-K-2Cl cotransport regulators on outflow facility in calf and human eyes in vitro

Lama A. Al-Aswad, Haiyan Gong, Lee David, Martha E O'Donnell, James D Brandt, William J. Ryan, Alison Schroeder, Kristine A. Erickson

Research output: Contribution to journalArticlepeer-review

42 Scopus citations


Purpose. Cultured human trabecular meshwork (TM) cells possess substantial Na-K-Cl activity, which is involved in the regulation of TM cell volume. The hypothesis in the present study was that drugs that affect the cotransporter might alter aqueous humor outflow facility (C) in the intact eye. The effects of agents and conditions known to modulate Na-K-Cl cotransport activity and/or TM cell volume on C in perfused anterior segments were investigated. Methods. Human and calf eyes were dissected and perfused, and C was determined according to standard published methods. Perfusates with modified osmolarity were used to cause alterations in TM cell volume. Cl- free perfusate and/or bumetanide (10-5 M) was used to inhibit Na-K-Cl cotransport activity, and vasopressin (10-7 M, 10-8 M) was used to stimulate cotransport activity. Results. In human eyes, hypo-osmotic perfusate decreased C 12%, whereas hyper-osmotic perfusate increased C 44%. These changes lasted approximately 30 minutes, after which C began to normalize. Inhibition of Na-K-Cl cotransport using Cl-free medium or bumetanide resulted in facility increases of 27% and 22%, respectively. There was an additive increase in C with bumetanide plus Cl-free media. Stimulating Na-K-Cl cotransport with 10-8 M and 10-7 M vasopressin resulted in 28% and 35% decreases in C, respectively. The results were similar in calf eyes: Cl-free medium or bumetanide resulted in 41% and 52% increases in C, whereas 10-8 M and 10-7 M vasopressin resulted in 14% and 19% decreases in C, respectively. Conclusions. Modulation of Na-K-Cl cotransport results in changes in C that may be mediated in part by cell volume changes.

Original languageEnglish (US)
Pages (from-to)1695-1701
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Issue number8
StatePublished - 1999

ASJC Scopus subject areas

  • Ophthalmology


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