AAV-Mediated, Optogenetic Ablation of Müller Glia Leads to Structural and Functional Changes in the Mouse Retina
Open Access
- 27 September 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (9), e76075
- https://doi.org/10.1371/journal.pone.0076075
Abstract
Müller glia, the primary glial cell in the retina, provide structural and metabolic support for neurons and are essential for retinal integrity. Müller cells are closely involved in many retinal degenerative diseases, including macular telangiectasia type 2, in which impairment of central vision may be linked to a primary defect in Müller glia. Here, we used an engineered, Müller-specific variant of AAV, called ShH10, to deliver a photo-inducibly toxic protein, KillerRed, to Müller cells in the mouse retina. We characterized the results of specific ablation of these cells on visual function and retinal structure. ShH10-KillerRed expression was obtained following intravitreal injection and eyes were then irradiated with green light to induce toxicity. Induction of KillerRed led to loss of Müller cells and a concomitant decrease of Müller cell markers glutamine synthetase and cellular retinaldehyde-binding protein, reduction of rhodopsin and cone opsin, and upregulation of glial fibrillary acidic protein. Loss of Müller cells also resulted in retinal disorganization, including thinning of the outer nuclear layer and the photoreceptor inner and outer segments. High resolution imaging of thin sections revealed displacement of photoreceptors from the ONL, formation of rosette-like structures and the presence of phagocytic cells. Furthermore, Müller cell ablation resulted in increased area and volume of retinal blood vessels, as well as the formation of tortuous blood vessels and vascular leakage. Electrophysiologic measures demonstrated reduced retinal function, evident in decreased photopic and scotopic electroretinogram amplitudes. These results show that loss of Müller cells can cause progressive retinal degenerative disease, and suggest that AAV delivery of an inducibly toxic protein in Müller cells may be useful to create large animal models of retinal dystrophies.Keywords
This publication has 28 references indexed in Scilit:
- Loss of Müller's Cells and Photoreceptors in Macular Telangiectasia Type 2Ophthalmology, 2013
- New functions of Müller cellsGlia, 2013
- Deleterious effects of mitochondrial ROS generated by KillerRed photodynamic action in human cell lines and C. elegansJournal of Photochemistry and Photobiology B: Biology, 2012
- Conditional Müller Cell Ablation Causes Independent Neuronal and Vascular Pathologies in a Novel Transgenic ModelJournal of Neuroscience, 2012
- Multifocal electroretinography in type 2 idiopathic macular telangiectasiaAlbrecht von Graefes Archiv für Ophthalmologie, 2012
- The IS/OS Junction Layer in the Natural History of Type 2 Idiopathic Macular TelangiectasiaInvestigative Ophthalmology & Visual Science, 2012
- Phototoxic effects of fluorescent protein KillerRed on tumor cells in miceJournal of Biophotonics, 2012
- Pilot Application of iTRAQ to the Retinal Disease Macular TelangiectasiaJournal of Proteome Research, 2011
- Intravitreal Injection of AAV2 Transduces Macaque Inner RetinaInvestigative Ophthalmology & Visual Science, 2011
- Submacular DL-α-Aminoadipic Acid Eradicates Primate Photoreceptors but Does Not Affect Luteal Pigment or the Retinal VasculatureInvestigative Ophthalmology & Visual Science, 2011