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Laser Capture Microdissection of Highly Pure Trabecular Meshwork from Mouse Eyes for Gene Expression Analysis
Published on: June 3, 2018
Cytoskeletal changes in cultured human glaucoma trabecular meshwork cells
A F Clark1, S T Miggans, K Wilson
1Alcon Laboratories, Fort Worth, Texas, U.S.A.
Purpose:
Glucocorticoid treatment of cultured human trabecular meshwork (TM) cells has been shown to reorganize actin microfilaments into cross-linked actin networks (CLANs) and to alter TM cell function. The purpose of the present study is to determine whether similar microfilament structural changes occur in TM cells derived from glaucoma patients.
Methods:
The microfilament structures of nine different TM cell cultures, derived from four glaucoma patients and from five normal subjects, was examined by epifluorescent microscopy and by whole-mount transmission electron microscopy. The cells were cultured in the absence and presence of dexamethasone (DEX) for 7-14 days.
Results:
The normal TM cell lines and glaucoma cell line TM13 had relatively low levels of cross-linked actin networks in the absence of exogenously added glucocorticoids. In contrast, the glaucoma cell lines TM23, TM36C, and TM48D had high levels of CLANs without the addition of dexamethasone. The addition of 10 M DEX to the culture medium significantly increased CLAN expression in all normal as well as glaucoma TM cell lines.
Conclusion:
CLAN formation in glaucoma TM cells or in TM cells exposed to glucocorticoids may be a factor contributing to the generation of ocular hypertension.
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