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B. T. Gabelt, J. Ruiz, R. Picciani, S. K. Bhattacharya, P. L. Kaufman; Cochlin Expression in MOCAS After TGFbeta2-Induced Elevated IOP. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4896.
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Determine whether cochlin, which is increased in ocular tissue in human glaucoma, is increased after transforming growth factor beta 2 (TGFß2)-induced intraocular pressure (IOP) elevation in monkey organ-cultured anterior segments (MOCAS) in vitro.
For MOCAS, rhesus monkey anterior segments were constantly infused with media (DMEM) containing 10ng/ml TGFß2 to one segment of each pair; 0.1% BSA to the opposite segment, at 2µl/min for up to 8 days. Media was collected and anterior segment quadrants were fixed or frozen for subsequent cochlin analysis. Outflow facility (OF) was measured by two-level constant pressure perfusion at baseline prior to treatment and at the final time point. Cochlin expression was determined by immunohistochemical and Western blot analysis of proteins extracted from MOCAS using chicken polyclonal antibodies against cochlin.
Two MOCAS infused with TGFß2 for 7-8 days developed elevated IOP (IOP increase of 25% and 100%). OF was decreased by approximately 30% compared to baseline. IOP did not change in two other MOCAS infused with TGFß2 for 2 days. Western blot of tissue extracts showed that cochlin levels were increased in MOCAS which developed elevated IOP as a result of TGFß2 treatment. Cochlin levels were also increased in one of the 2 segments infused with TGFß2 for only 2 days. Cochlin was not detected in any of the 4 vehicle treated MOCAS. Immunohistochemical analysis of tissue from MOCAS with elevated IOP showed increased cochlin expression in the trabecular meshwork and remnants of the ciliary body.
Cochlin expression was found to be increased in MOCAS in which IOP was elevated as a result of TGFß2 treatment. Cochlin over-expression may also precede IOP elevation. Whether cochlin over-expression contributes to elevated IOP or is a consequence of changes in other factors that lead to elevated IOP remains to be determined.
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