Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration

We aimed to verify whether the intravitreal injection of small molecule compounds alone can create photoreceptor cells in mouse models of retinal degeneration. Primary cultured mouse Müller cells were stimulated in vitro with combinations of candidate compounds and the rhodopsin expression was measu...

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Autores principales: Fujii, Yuya, Arima, Mitsuru, Murakami, Yusuke, Sonoda, Koh-Hei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949636/
https://www.ncbi.nlm.nih.gov/pubmed/36821627
http://dx.doi.org/10.1371/journal.pone.0282174
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author Fujii, Yuya
Arima, Mitsuru
Murakami, Yusuke
Sonoda, Koh-Hei
author_facet Fujii, Yuya
Arima, Mitsuru
Murakami, Yusuke
Sonoda, Koh-Hei
author_sort Fujii, Yuya
collection PubMed
description We aimed to verify whether the intravitreal injection of small molecule compounds alone can create photoreceptor cells in mouse models of retinal degeneration. Primary cultured mouse Müller cells were stimulated in vitro with combinations of candidate compounds and the rhodopsin expression was measured on day 7 using polymerase chain reaction and immunostaining. We used 6-week-old N-methyl-N-nitrosourea-treated and 4-week-old rd10 mice as representative in vivo models of retinal degeneration. The optimal combination of compounds selected via in vitro screening was injected into the vitreous and the changes in rhodopsin expression were investigated on day 7 using polymerase chain reaction and immunostaining. The origin of rhodopsin-positive cells was also analyzed via lineage tracing and the recovery of retinal function was assessed using electroretinography. The in vitro mRNA expression of rhodopsin in Müller cells increased 30-fold, and 25% of the Müller cells expressed rhodopsin protein 7 days after stimulation with a combination of 4 compounds: transforming growth factor-β inhibitor, bone morphogenetic protein inhibitor, glycogen synthase kinase 3 inhibitor, and γ-secretase inhibitor. The in vivo rhodopsin mRNA expression and the number of rhodopsin-positive cells in the outer retina were significantly increased on day 7 after the intravitreal injection of these 4 compounds in both N-methyl-N-nitrosourea-treated and rd10 mice. Lineage tracing in td-Tomato mice treated with N-methyl-N-nitrosourea suggested that the rhodopsin-positive cells originated from endogenous Müller cells, accompanied with the recovery of the rhodopsin-derived scotopic function. It was suggested that rhodopsin-positive cells generated by compound stimulation contributes to the recovery of retinal function impaired by degeneration.
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spelling pubmed-99496362023-02-24 Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration Fujii, Yuya Arima, Mitsuru Murakami, Yusuke Sonoda, Koh-Hei PLoS One Research Article We aimed to verify whether the intravitreal injection of small molecule compounds alone can create photoreceptor cells in mouse models of retinal degeneration. Primary cultured mouse Müller cells were stimulated in vitro with combinations of candidate compounds and the rhodopsin expression was measured on day 7 using polymerase chain reaction and immunostaining. We used 6-week-old N-methyl-N-nitrosourea-treated and 4-week-old rd10 mice as representative in vivo models of retinal degeneration. The optimal combination of compounds selected via in vitro screening was injected into the vitreous and the changes in rhodopsin expression were investigated on day 7 using polymerase chain reaction and immunostaining. The origin of rhodopsin-positive cells was also analyzed via lineage tracing and the recovery of retinal function was assessed using electroretinography. The in vitro mRNA expression of rhodopsin in Müller cells increased 30-fold, and 25% of the Müller cells expressed rhodopsin protein 7 days after stimulation with a combination of 4 compounds: transforming growth factor-β inhibitor, bone morphogenetic protein inhibitor, glycogen synthase kinase 3 inhibitor, and γ-secretase inhibitor. The in vivo rhodopsin mRNA expression and the number of rhodopsin-positive cells in the outer retina were significantly increased on day 7 after the intravitreal injection of these 4 compounds in both N-methyl-N-nitrosourea-treated and rd10 mice. Lineage tracing in td-Tomato mice treated with N-methyl-N-nitrosourea suggested that the rhodopsin-positive cells originated from endogenous Müller cells, accompanied with the recovery of the rhodopsin-derived scotopic function. It was suggested that rhodopsin-positive cells generated by compound stimulation contributes to the recovery of retinal function impaired by degeneration. Public Library of Science 2023-02-23 /pmc/articles/PMC9949636/ /pubmed/36821627 http://dx.doi.org/10.1371/journal.pone.0282174 Text en © 2023 Fujii et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Fujii, Yuya
Arima, Mitsuru
Murakami, Yusuke
Sonoda, Koh-Hei
Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title_full Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title_fullStr Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title_full_unstemmed Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title_short Rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
title_sort rhodopsin-positive cell production by intravitreal injection of small molecule compounds in mouse models of retinal degeneration
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949636/
https://www.ncbi.nlm.nih.gov/pubmed/36821627
http://dx.doi.org/10.1371/journal.pone.0282174
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