Table 1B.
Selected Trabecular Meshwork Functions Altered by Wild-Type and Mutant Myocilin
| Function | Model | Method | Vector | Species | Citation |
|---|---|---|---|---|---|
| Wild-type myocilin | |||||
| Loss of stress fibers | Transi.transf.HTM/ HVJ liposome/ 48h | Immunofluoresc.Phalloidin | pcDNA3.1.myoc | Human | Wentz-Hunter 2004 |
| Stably transf.HTM cells/ FuGene | Immunofluoresc.Phalloidin | Human | |||
| Induction stress fibers | Exogen myoc on HTM / 3h | Immunofluoresc.Phalloidin | purified myocilin | Human | Kwon 2009 |
| from transfec 293 media | |||||
| β-Catenin translocation | Exogenous myoc on NIH3T3 cells | Immunofluoresc.βcatenin | from transfec 293 media | Mouse | Kwon 2009 |
| Wnt pathway activation | Tcf/Lef binding site/ luciferase reporter | Chemiluminescence | Myoc+pTOP-Flash | Human | Shen 2012 |
| Transf.Colorectal adenocarcinoma cell | |||||
| Reduce RhoA activation | Transi.transf.HTM/ FuGene&Lipof.LTX | Pull-down assay | pTarget.myocilin | Human | Shen 2008 |
| Reduce fibronectin deposition | Transi.transf.HTM | Fluores. Histocytochem. | pTarget.myocilin | Human | Shen 2008 |
| Higher sensitivity to apoptosis | Transi.transf.HTM/ HVJ liposome/ 48h | Anti-Fas apoptosis inducti | pcDNA3.1.myoc | Human | Wentz-Hunter 2004 |
| Conversion to osteoblasts | Mesenchimal stem.cells (MSC) | AlizaRed stain./Alkal.Phos. | purified myocilin | Human | Kwon 2013 |
| 3μg myoc for 6d | from transfec 293 media | ||||
| Inhibits cell spreading | Hskin fibroblast on coated myoc well | Count spread/round cells | Hmyoc fr transf insectcells | Human | Peters 2005 |
| Inhibits incorp Paxillin in FA | Hskin fibrobroblast+soluble myocilin | Immunofluoresc. Vin&paxil | Hmyoc fr transf insectcells | Human | Peters 2005 |
| Mutant myocilin | |||||
| Disturbs ER stress response | Transi.transf.HTM/ Lipofectmine | pcr.GRP78 & P.eIF2a | pCDNA3.P370L | Human | Wang 2007 |
| Disturbs Mitchon membr potential | Transi.transf.HTM/ Lipofectmine | Flow cytometry. JC1probe | pCDNA3.P370L | Human | Wang 2007 |
| Loss of stress fibers | Transi.transf.HTM/ FuGene&Lipof.LTX | FuGene & Lipofecta.LTX | pMyoc.P370L-EGFP | Human | Shen 2012 |
| Increase prot kinA (PKA) activitity | Transi.transf.HTM/ FuGene&Lipof.LTX | Peptag L-R-R-S-L-G | pMyoc.P370L-EGFP | Human | Shen 2012 |
| Reduce RhoA activation | Transi.transf.HTM/ FuGene&Lipof.LTX | Pull-down assay | pMyoc.P370L-EGFP | Human | Shen 2012 |
| Incre sensitivity to oxidative stress | HEK 293*/ 100μM H2O2 | Apoptosis/ TUNEL | pTRE-Y437H MYOC | Human | Joe 2010 |
| Mild glaucoma | Transg.mice/ hmyoc | IOP/ RGC&Axon count | BAC RP11-23L17.Y437H | Mouse | Zhou 2008 |
| Glaucoma | Transg.mice/ CMV.cDNAhmyoc | IOP/ RGC&Axon count | pCS2.Y437H | Mouse | Zode 2011 |
| Inhibition of myoc processing | Transi.transf. HEK293, COS, hCMsv | WB | pCDNA3.D380A | H &Monk | Aroca-Aguilar 2005 |
HEK 293*, HEK 293 Tet-On stably transfected.
HTM, primary human trabecular meshwork cells; ICC, immunocytochemistry; FA, focal adhesion; ER, endoplasmic reticulum; IOP, intraocular pressure.