Segmental versican expression in the trabecular meshwork and involvement in outflow facility
| Autor: | Ted S. Acott, John M. Bradley, Kate E. Keller, Janice A. Vranka |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Swine
Genetic Vectors Gene Expression Glycosaminoglycan Aqueous Humor chemistry.chemical_compound Versicans Trabecular Meshwork Quantum Dots medicine Gene silencing Animals Humans Chondroitin sulfate Gene Silencing RNA Messenger Fluorescent Antibody Technique Indirect Cells Cultured Messenger RNA Microscopy Confocal biology Reverse Transcriptase Polymerase Chain Reaction Lentivirus Articles Molecular biology carbohydrates (lipids) medicine.anatomical_structure chemistry Proteoglycan Gene Expression Regulation biology.protein cardiovascular system Versican RNA Interference Trabecular meshwork sense organs Immunostaining |
| Zdroj: | Investigative ophthalmologyvisual science. 52(8) |
| ISSN: | 1552-5783 |
| Popis: | PURPOSE. Versican is a large proteoglycan with numerous chondroitin sulfate (CS) glycosaminoglycan (GAG) side chains attached. To assess versican’s potential contributions to aqueous humor outflow resistance, its segmental distribution in the trabecular meshwork (TM) and the effect on outflow facility of silencing the versican gene were evaluated. METHODS. Fluorescent quantum dots (Qdots) were perfused to label outflow pathways of anterior segments. Immunofluorescence with confocal microscopy and quantitative RT-PCR were used to determine versican protein and mRNA distribution relative to Qdot-labeled regions. Lentiviral delivery of shRNAsilencing cassettes to TM cells in perfused anterior segment cultures was used to evaluate the involvement of versican and CS GAG chains in outflow facility. RESULTS. Qdot uptake by TM cells showed considerable segmental variability in both human and porcine outflow pathways. Regional levels of Qdot labeling were inversely related to versican protein and mRNA levels; versican levels were relatively high in sparsely Qdot-labeled regions and low in densely labeled regions. Versican silencing decreased outflow facility in human and increased facility in porcine anterior segments. However, RNAi silencing of ChGn, an enzyme unique to CS GAG biosynthesis, increased outflow facility in both species. The fibrillar pattern of versican immunostaining in the TM juxtacanalicular region was disrupted after versican silencing in perfusion culture. CONCLUSIONS. Versican appears to be a central component of the outflow resistance, where it may organize GAGs and other ECM components to facilitate and control open flow channels in the TM. However, the exact molecular organization of this resistance appears to differ between human and porcine eyes. (Invest Ophthalmol Vis Sci. 2011;52:5049 –5057) DOI |
| Databáze: | OpenAIRE |
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