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Robin R. Hodges, Joanna Vrouvlianis, Marie A. Shatos, Darlene A. Dartt; Characterization of P2X7 Purinergic Receptors and Their Function in Rat Lacrimal Gland. Invest. Ophthalmol. Vis. Sci. 2009;50(12):5681-5689. doi: 10.1167/iovs.09-3670.
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To characterize the effects of P2X7 purinergic receptors on lacrimal gland function.
P2X7 purinergic receptors were identified by RT-PCR, Western blot analysis, and immunofluorescence techniques. Rat lacrimal gland acini were isolated by collagenase digestion. Acini were incubated with the fluorescent indicator molecule fura 2, and [Ca2+]i was measured by a fluorescence imaging system. Protein secretion was measured with a fluorescence assay system. Activation of ERK 1/2 was determined by Western blot analysis with an antibody against phosphorylated (active) ERK 1/2.
P2X7 receptors were present in the lacrimal gland by RT-PCR and Western blot analysis. These receptors were located in the membranes of acinar and ductal cells and the cytoplasm of acinar cells. Activation of P2X7 receptors with (benzoylbenzoyl)adenosine 5′-triphosphate increased [Ca2+]i, peroxidase secretion, and ERK 1/2 activation, each of which was inhibited by the P2X7 receptor inhibitors Brilliant Blue G or A 438079.
P2X7 purinergic receptors are present in rat lacrimal gland and when stimulated increase [Ca2+]i, protein secretion, and ERK 1/2 activation.
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