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Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71
PURPOSE: To study the potential effect of a gene therapy, designed to rescue the expression of dystrophin Dp71 in the retinas of Dp71-null mice, on retinal physiology. METHODS: We recorded electroretinograms (ERGs) in Dp71-null and wild-type littermate mice. In dark-adapted eyes, responses to flashe...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The Association for Research in Vision and Ophthalmology
2020
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7326481/ https://www.ncbi.nlm.nih.gov/pubmed/32049345 http://dx.doi.org/10.1167/iovs.61.2.11 |
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author | Barboni, Mirella Telles Salgueiro Vaillend, Cyrille Joachimsthaler, Anneka Liber, André Maurício Passos Khabou, Hanen Roux, Michel J. Vacca, Ophélie Vignaud, Lucile Dalkara, Deniz Guillonneau, Xavier Ventura, Dora Fix Rendon, Alvaro Kremers, Jan |
author_facet | Barboni, Mirella Telles Salgueiro Vaillend, Cyrille Joachimsthaler, Anneka Liber, André Maurício Passos Khabou, Hanen Roux, Michel J. Vacca, Ophélie Vignaud, Lucile Dalkara, Deniz Guillonneau, Xavier Ventura, Dora Fix Rendon, Alvaro Kremers, Jan |
author_sort | Barboni, Mirella Telles Salgueiro |
collection | PubMed |
description | PURPOSE: To study the potential effect of a gene therapy, designed to rescue the expression of dystrophin Dp71 in the retinas of Dp71-null mice, on retinal physiology. METHODS: We recorded electroretinograms (ERGs) in Dp71-null and wild-type littermate mice. In dark-adapted eyes, responses to flashes of several strengths were measured. In addition, flash responses on a 25-candela/square meters background were measured. On- and Off-mediated responses to sawtooth stimuli and responses to photopic sine-wave modulation (3–30 Hz) were also recorded. After establishing the ERG phenotype, the ShH10-GFP adeno-associated virus (AAV), which has been previously shown to target specifically Müller glial cells (MGCs), was delivered intravitreously with or without (sham therapy) the Dp71 coding sequence under control of a CBA promoter. ERG recordings were repeated three months after treatment. Real-time quantitative PCR and Western blotting analyses were performed in order to quantify Dp71 expression in the retinas. RESULTS: Dp71-null mice displayed reduced b-waves in dark- and light-adapted flash ERGs and smaller response amplitudes to photopic rapid-on sawtooth modulation and to sine-wave stimuli. Three months after intravitreal injections of the ShH10-GFP-2A-Dp71 AAV vector, ERG responses were completely recovered in treated eyes of Dp71-null mice. The functional rescue was associated with an overexpression of Dp71 in treated retinas. CONCLUSIONS: The present results show successful functional recovery accompanying the reexpression of Dp71. In addition, this experimental model sheds light on MGCs influencing ERG components, since previous reports showed that aquaporin 4 and Kir4.1 channels were mislocated in MGCs of Dp71-null mice, while their distribution could be normalized following intravitreal delivery of the same ShH10-GFP-2A-Dp71 vector. |
format | Online Article Text |
id | pubmed-7326481 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | The Association for Research in Vision and Ophthalmology |
record_format | MEDLINE/PubMed |
spelling | pubmed-73264812020-07-07 Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 Barboni, Mirella Telles Salgueiro Vaillend, Cyrille Joachimsthaler, Anneka Liber, André Maurício Passos Khabou, Hanen Roux, Michel J. Vacca, Ophélie Vignaud, Lucile Dalkara, Deniz Guillonneau, Xavier Ventura, Dora Fix Rendon, Alvaro Kremers, Jan Invest Ophthalmol Vis Sci Visual Neuroscience PURPOSE: To study the potential effect of a gene therapy, designed to rescue the expression of dystrophin Dp71 in the retinas of Dp71-null mice, on retinal physiology. METHODS: We recorded electroretinograms (ERGs) in Dp71-null and wild-type littermate mice. In dark-adapted eyes, responses to flashes of several strengths were measured. In addition, flash responses on a 25-candela/square meters background were measured. On- and Off-mediated responses to sawtooth stimuli and responses to photopic sine-wave modulation (3–30 Hz) were also recorded. After establishing the ERG phenotype, the ShH10-GFP adeno-associated virus (AAV), which has been previously shown to target specifically Müller glial cells (MGCs), was delivered intravitreously with or without (sham therapy) the Dp71 coding sequence under control of a CBA promoter. ERG recordings were repeated three months after treatment. Real-time quantitative PCR and Western blotting analyses were performed in order to quantify Dp71 expression in the retinas. RESULTS: Dp71-null mice displayed reduced b-waves in dark- and light-adapted flash ERGs and smaller response amplitudes to photopic rapid-on sawtooth modulation and to sine-wave stimuli. Three months after intravitreal injections of the ShH10-GFP-2A-Dp71 AAV vector, ERG responses were completely recovered in treated eyes of Dp71-null mice. The functional rescue was associated with an overexpression of Dp71 in treated retinas. CONCLUSIONS: The present results show successful functional recovery accompanying the reexpression of Dp71. In addition, this experimental model sheds light on MGCs influencing ERG components, since previous reports showed that aquaporin 4 and Kir4.1 channels were mislocated in MGCs of Dp71-null mice, while their distribution could be normalized following intravitreal delivery of the same ShH10-GFP-2A-Dp71 vector. The Association for Research in Vision and Ophthalmology 2020-02-12 2020-02 /pmc/articles/PMC7326481/ /pubmed/32049345 http://dx.doi.org/10.1167/iovs.61.2.11 Text en Copyright 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. |
spellingShingle | Visual Neuroscience Barboni, Mirella Telles Salgueiro Vaillend, Cyrille Joachimsthaler, Anneka Liber, André Maurício Passos Khabou, Hanen Roux, Michel J. Vacca, Ophélie Vignaud, Lucile Dalkara, Deniz Guillonneau, Xavier Ventura, Dora Fix Rendon, Alvaro Kremers, Jan Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title | Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title_full | Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title_fullStr | Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title_full_unstemmed | Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title_short | Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71 |
title_sort | rescue of defective electroretinographic responses in dp71-null mice with aav-mediated reexpression of dp71 |
topic | Visual Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7326481/ https://www.ncbi.nlm.nih.gov/pubmed/32049345 http://dx.doi.org/10.1167/iovs.61.2.11 |
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