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(Investigative Ophthalmology and Visual Science. 2008;49:5517-5525.)
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-0435
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Role of Chloride Channels in Regulating the Volume of Acinar Cells of the Rabbit Superior Lacrimal Gland

George H. Herok,1,2 Thomas J. Millar,1,2 Philip J. Anderton,1 and Donald K. Martin3

1From the Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Kensington, NSW, Australia; the 2Department of Natural Sciences, University of Western Sydney/Nepean, Sydney, NSW, Australia; and the 3Department of Medical and Molecular Biology, Faculty of Science, University of Technology, Sydney, NSW, Australia.

PURPOSE. To characterize the outward chloride currents (ClOR) in single acinar cells isolated from the rabbit superior lacrimal gland (RSLG) to investigate the hypothesis that ClOR may have a role in regulating the volume of RSLG acini.

METHODS. ClOR was characterized by using patch-clamp electrophysiology. Confocal microscopy was used to measure intracellular calcium concentration ([Ca2+]i) and cell volume. Cell volume was altered by superfusing the cells with a hyposmotic solution.

RESULTS. The ClOR current contributed 33% of total membrane conductance. With normal osmotic conditions, the ClOR current was activated by [Ca2+]i with an EC50 of 10–8 M. A decrease in intracellular pH from 7.4 to 6.8 totally inhibited ClOR current activity. Continuous superfusion of hyposmotic solution caused (1) an increase in cell volume that peaked within 4 minutes and gradually returned to baseline levels after 12 minutes, (2) an increase in [Ca2+]i that peaked between 6 and 8 minutes and gradually returned to baseline levels after 15 minutes, and (3) an increase the ClOR current that peaked within 6 minutes after commencement of perfusion and quickly returned to baseline levels.

CONCLUSIONS. The ClOR current appears to be triggered by an increase in cell volume and then deactivates within the period of raised [Ca2+]i during hyposmotic stress, suggesting that ClOR may be an initiating event for volume homeostasis. This effect would be important during RSLG tear secretion, which usually involves cell volume changes and is accompanied by intracellular pH changes in the presence of the raised [Ca2+]i to support secretion.






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