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Xiaofang Wang, Douglas H. Johnson; mRNA In Situ Hybridization of TIGR/MYOC in Human Trabecular Meshwork. Invest. Ophthalmol. Vis. Sci. 2000;41(7):1724-1729.
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purpose. To determine the distribution of mRNA expression of the trabecular
meshwork–induced glucocorticoid response protein/myocilin (TIGR/MYOC)
in human trabecular meshwork.
methods. In situ hybridization using a 1.25-kb probe obtained from reverse
transcription–polymerase chain reaction of TIGR/MYOC cDNA was performed to determine
the location of cell labeling within the different regions of the
meshwork. The effect of dexamethasone on the pattern of labeling was
studied in organ cultured meshwork. Trabecular meshwork from three
sources was studied: enucleated eyes obtained at autopsy,
trabeculectomy specimens obtained during filtration surgery, and
meshworks from anterior segments in perfusion organ culture.
Hybridization was performed on frozen sections, paraffin sections, and
sections from JB-4 plastic–embedded tissue.
results. Labeling for TIGR/MYOC mRNA was present in most
trabecular cells of the uveal, corneoscleral, and juxtacanalicular
regions but only variably present in the endothelial cells of
Schlemm’s canal. A similar pattern was found in the trabeculectomy
specimens from eyes with primary open-angle or pseudoexfoliative
glaucoma. Dexamethasone treatment increased the labeling intensity and
number of labeled cells in meshwork, and also the number of labeled
endothelial cells of Schlemm’s canal. Fresh tissue processed within 12
hours postmortem gave more consistent labeling than older tissue,
although some label was found up to 48 hours postmortem. Labeling was
found in tissue from all three sources, and with all three embedding
techniques; JB-4 sections provided the best morphologic resolution.
conclusions. In situ hybridization reveals that mRNA expression for TIGR/MYOC is present in most cells in all regions of the
meshwork but only variably present in the endothelial cells of
Schlemm’s canal. Dexamethasone treatment increased the number and
intensity of labeled cells, and also increased the number of labeled
cells in the endothelial lining of Schlemm’s
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