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(Investigative Ophthalmology and Visual Science. 2007;48:3746-3755.)
© 2007 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.06-1291
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New Experimental Method to Study Acid/Base Transporters and Their Regulation in Lacrimal Gland Ductal Epithelia

Edit Tóth-Molnár,1,2 Viktória Venglovecz,2,3 Béla Ózsvári,3 Zoltán Rakonczay, Jr,3 András Varró,4,5 Julius G. Papp,4,5 András Tóth,4 János Lonovics,3 Tamás Takács,3 Imre Ignáth,3 Béla Iványi,6 and Péter Hegyi3,4

1From the Departments of Ophthalmology, 4Pharmacology and Pharmacotherapy, and 6Pathology, and the 3First Department of Medicine, Faculty of Medicine, University of Szeged, Szeged, Hungary; and the 5Division of Cardiovascular Pharmacology, Hungarian Academy of Sciences, Szeged, Hungary.

PURPOSE. The main function of the lacrimal gland is to produce the most aqueous component of the tear film covering the surfaces of the cornea and the conjunctiva. Studies have been conducted that characterize the mixed fluid and protein secretion of isolated acini, but no methods have been developed to characterize lacrimal gland ductal cell (LGDC) secretion. Secretory mechanisms of ductal epithelia may play physiological roles in the maintenance of the standard environments for the cornea and the conjunctiva.

METHODS. In this study, the authors developed a rapid method to isolate large quantities of intact lacrimal ducts. The preparation of isolated intact lacrimal gland ducts for the first time enabled the performance of real-time functional experiments on cleaned ducts. Electron microscopy and fluorescence measurements were used to evaluate the viability of lacrimal ducts.

RESULTS. Fluorescence measurements showed that LGDCs express functionally active Na+/H+ exchanger (NHE) and Cl–/HCO3– exchanger (AE). Parasympathomimetic stimulation by carbachol stimulated NHE and AE through the elevation of intracellular calcium concentration. This mechanism can play a role in the regulation of ion and water secretion by LGDCs.

CONCLUSIONS. The authors have described a lacrimal gland duct isolation technique in which the intact ducts remain viable and the role of duct cells in tear film secretion can be characterized. These data combined with the novel isolation facilitated understanding of the regulation mechanisms of ductal cell secretion at cellular and molecular levels under normal and pathologic conditions.






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