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(Investigative Ophthalmology and Visual Science. 2004;45:3974-3981.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0508
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Effect of Overexpression of Constitutively Active PKC{alpha} on Rat Lacrimal Gland Protein Secretion

Robin R. Hodges,1 Isabelle Raddassi,1 Driss Zoukhri,2 Alex Toker,3 Andrius Kazlauskas,1 and Darlene A. Dartt1

1From the Schepens Eye Research Institute and Department of Ophthalmology, and the 3Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts; and the 2Tufts University School of Dental Medicine, Boston, Massachusetts.

PURPOSE. The lacrimal gland secretes water, electrolytes, and protein into the tear film. Decreased secretion from the lacrimal gland can lead to dry eye syndromes with deleterious effects on vision. Protein kinase C (PKC)-{alpha} plays a major role in cholinergic- and {alpha}1-adrenergic–induced protein secretion from the lacrimal gland. This study was undertaken to determine whether activation of PKC{alpha} alone would induce lacrimal gland protein secretion by examining the effects of overexpression of constitutively active PKC{alpha}.

METHODS. Rat lacrimal gland acini were transduced with an adenovirus containing a gene for constitutively active PKC{alpha}. Protein secretion was measured in response to cholinergic and {alpha}1-adrenergic agonist stimulation.

RESULTS. More than 84% of acinar cells were transduced, and PKC{alpha} expression was increased 176-fold. Western blot analysis using an antibody to phosphorylated (activated) PKC{alpha} indicated that the overexpressed PKC{alpha} was active, and basal secretion was increased. Cholinergic agonist–stimulated protein secretion was not stimulated above basal secretion, whereas {alpha}1-adrenergic-agonist–stimulated protein secretion was increased in transduced acini.

CONCLUSIONS. Basal lacrimal gland protein secretion can be stimulated by bypassing the release of neurotransmitters and activating PKC{alpha}, possibly leading to the development of new treatments for dry eye syndromes.






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