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D. Nguyen, T.L. Nhan, H. Toshida, L.P. Pedroza, R.W. Beuerman; Loss of Muscarinic, Parasympathetic Activation Induces Long–Term Changes in Proinflammatory and Immune Response Gene Expression in the Rat Lacrimal Gland . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2418.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To examine long–term loss of parasympathetic activation of secretion on the expression of proinflammatory mediators and the immune status in the rat lacrimal gland. Methods: Rats underwent unilateral pre–ganglionic parasympathetic sectioning of the greater superficial petrosal nerve and were sacrificed after 2.5 months. The structure of the LG acini was examined by light microscopy. The LG from the contralateral (Ctla) and parasympathectomized (Px) side was used for real–time RT–PCR (n=3). All reactions were done in triplicate using the iCycler IQ real–time detection system. Mean cycle threshold (MCt) for the genes in the Ctla and Px LG were normalized to 18S rRNA MCt values. Relative fold change was calculated for each of the genes using the 2–ΔΔCT method and compared to the expression level after 7 days. Results:Light microscopy showed an increase in the number of seromucous granules and apoptotic cells in the Px LG. Expression of caspase 1, NFΚB p105, and MMP–12 remained elevated (greater than 2–fold) in the Px LG after 2.5 months. Expression of the MHC class II antigen RT1.B (HLA–DQB orthologue) and RT1.D (HLA–DRA orthologue), increased from 2.0– and 2.4–fold after 7 days, to 7.6– and 6.3–fold after 2.5 months, respectively. In addition, expression of Psmb8, a proteasome subunit RC1 involved in peptide processing, increased from 1.4–fold after 7 days, to 4.6–fold after 2.5 months. Conversely, expression of the ER–resident protein, protein disulphide isomerase, which showed a 2–fold decreased after 7 days, was not altered after 2.5 months. The MCt of the 18S rRNA (Mean + SEM) was not significantly different between Ctla (11.70 + 0.00) and Px LGs (11.63 + 0.04). Conclusions: Removal of parasympathetic control of secretion leads to a chronic and persistent upregulation of the immune response in the lacrimal gland. These changes correlate with age–associated changes in the LG.
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