A Novel Mouse Model of TGFβ2-Induced Ocular Hypertension Using Lentiviral Gene Delivery
Abstract
:1. Introduction
2. Results
2.1. Intravitreal Injection of Lentiviral Vector Expressing hTGFβ2C226/228S Leads to Sustained IOP Elevation in Mice
2.2. Decrease of Aqueous Humor Outflow Facility in LV_TGFβ2 Expressing Eyes
2.3. Gross Morphological Assessment of Ocular Tissues in LV-Injected Mice
2.4. TGFβ2 Expression in AH and Anterior Segment Tissue of LV_TGFβ2 Injected Eyes
2.5. LV_TGFβ2 Stimulates ECM Deposition and Actin Reorganization in Primary TM Cells
3. Discussion
4. Methods and Materials
4.1. Mice
4.2. Viral Vector Injections
4.3. IOP Measurements
4.4. Outflow Facility Measurements
4.5. Ophthalmoscopy and Slit-Lamp Examination
4.6. Ocular Sample Collection
4.7. Lentiviral Transduction of Primary Human Trabecular Meshwork Cells
4.8. H&E Staining
4.9. Immunostaining
4.10. Western Blotting
4.11. Pattern Electroretinography
4.12. RGC Counts from Retinal Whole Mounts
4.13. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibodies | Catalog # | Company | Dilutions |
---|---|---|---|
Transforming Growth Factor-β2 | Ab36495 | Abcam, Cambridge, MA, USA | 1:100 |
Fibronectin (FN) | Ab2413 | Abcam | 1:400 |
Collagen-1 (Col-1) | NB600-408 | Novus Biologicals, Littleton, CO, USA | 1:100 |
α-Smooth Muscle Actin (αSMA) | Ab7817 | Abcam | 1:100 |
Phalloidin-Alexa Conjugate | 8878S | Cell Signaling Technology, Danvers, MA, USA | 1:20 |
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Patil, S.V.; Kasetti, R.B.; Millar, J.C.; Zode, G.S. A Novel Mouse Model of TGFβ2-Induced Ocular Hypertension Using Lentiviral Gene Delivery. Int. J. Mol. Sci. 2022, 23, 6883. https://doi.org/10.3390/ijms23136883
Patil SV, Kasetti RB, Millar JC, Zode GS. A Novel Mouse Model of TGFβ2-Induced Ocular Hypertension Using Lentiviral Gene Delivery. International Journal of Molecular Sciences. 2022; 23(13):6883. https://doi.org/10.3390/ijms23136883
Chicago/Turabian StylePatil, Shruti V., Ramesh B. Kasetti, J. Cameron Millar, and Gulab S. Zode. 2022. "A Novel Mouse Model of TGFβ2-Induced Ocular Hypertension Using Lentiviral Gene Delivery" International Journal of Molecular Sciences 23, no. 13: 6883. https://doi.org/10.3390/ijms23136883