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(Investigative Ophthalmology and Visual Science. 2006;47:794-801.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-0292
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Biochemical and Morphological Analysis of Basement Membrane Component Expression in Corneoscleral and Cribriform Human Trabecular Meshwork Cells

Rudolf Fuchshofer,1 Ulrich Welge-Lüssen,2 Elke Lütjen-Drecoll,3 and Marco Birke3

1From the Department of Human Anatomy, University Regensburg, Regensburg, Germany; the 2Department of Ophthalmology, Maximilians-University, Munich, Germany; and the 3Department of Anatomy II, Friedrich Alexander University, Erlangen, Germany.

PURPOSE. To determine whether cells of the cribriform trabecular meshwork (TM) express basement membrane (BM) components similar to corneoscleral TM cells and to determine whether cribriform cells are connected to the elastic tendon net of the TM.

METHODS. TM cells of the corneoscleral and the cribriform regions were cultured from 10 eyes of 10 donors, aged 20 to 87 years. Cell types were classified by {alpha}-smooth muscle actin (smA), desmin, and {alpha}B-crystallin staining. Expression of collagen type IV (ColIV) chains {alpha}1 to 6; collagen type VIII (ColVIII) {alpha}1; laminin subunits {alpha}1 to 5, ß1 to 3, and {gamma}1 to 3; and nidogen 1 and 2 was tested in both cell types by semiquantitative RT-PCR (sqPCR). Expression of ColIV{alpha}2, ColVIII{alpha}1, laminin ß2, and nidogen 1 was quantified by Northern blot analysis. The response to transforming growth factor (TGF)-ß2 treatment was investigated. Serial tangential and sagittal TM sections of 16 eyes from 10 donors (aged 12–90 years) were used for electron- and immunoelectron microscopy.

RESULTS. Both TM cell types expressed ColIV chains {alpha}1, {alpha}2, {alpha}4, {alpha}5, and {alpha}6; ColVIII {alpha}1; laminin subunits {alpha}3, {alpha}4, ß1, ß2, ß3, {gamma}1, and {gamma}2; and nidogen 1, as determined by Northern blot analysis and sqPCR. ColIV {alpha}3; laminin subunits {alpha}1, {alpha}2, and {gamma}3; and nidogen 2 were not detectable by PCR. Responses to TGF-ß2 treatment did not differ between cell types. In vivo, all cribriform cells were in contact with ColIV containing BM material and were found to connect to the cribriform elastic network.

CONCLUSIONS. Cribriform and corneoscleral TM cells show no differences in expression of BM components and response to TGF-ß2. The direct connection of cribriform cells to the elastic tendon network suggests that they are under mechanical tension. This could explain previous findings of {alpha}B-crystallin expression in the cribriform region.




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