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The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina

PURPOSE: Because the importance of glia in regulating brain functions has been demonstrated, genetic technologies that manipulate glial cell-specific gene expression in the brain have become essential and have made great progress. However, it is unknown whether the same strategy that is used in the...

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Autores principales: Danjo, Yosuke, Shinozaki, Youichi, Natsubori, Akiyo, Kubota, Yuto, Kashiwagi, Kenji, Tanaka, Kenji F., Koizumi, Schuichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Association for Research in Vision and Ophthalmology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8764212/
https://www.ncbi.nlm.nih.gov/pubmed/35040915
http://dx.doi.org/10.1167/tvst.11.1.25
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author Danjo, Yosuke
Shinozaki, Youichi
Natsubori, Akiyo
Kubota, Yuto
Kashiwagi, Kenji
Tanaka, Kenji F.
Koizumi, Schuichi
author_facet Danjo, Yosuke
Shinozaki, Youichi
Natsubori, Akiyo
Kubota, Yuto
Kashiwagi, Kenji
Tanaka, Kenji F.
Koizumi, Schuichi
author_sort Danjo, Yosuke
collection PubMed
description PURPOSE: Because the importance of glia in regulating brain functions has been demonstrated, genetic technologies that manipulate glial cell-specific gene expression in the brain have become essential and have made great progress. However, it is unknown whether the same strategy that is used in the brain can be applied to the retina because retinal glia differs from glia in the brain. Here, we aimed to find a method for selective gene expression in Müller cells (characteristic glial cells in the retina) and identified Mlc1 as a specific promoter of Müller cells. METHODS: Mlc1-tTA::Yellow-Cameleon-Nano(tetO/tetO) (YC-Nano) mice were used as a reporter line. YC-Nano, a fluorescent protein, was ectopically expressed in the cell type controlled by the Mlc1 promotor. Immunofluorescence staining was used to identify the cell type expressing YC-Nano protein. RESULTS: YC-Nano-positive ((+)) signals were observed as vertical stalks in the sliced retina and spanned from the nerve fiber layer through the outer nuclear layer. The density of YC-Nano(+) cells was higher around the optic nerve head and lower in the peripheral retina. The YC-Nano(+) signals colocalized with vimentin, a marker of Müller cells, but not with the cell markers for blood vessels, microglia, neurons, or astrocytes. CONCLUSIONS: The Mlc1 promoter allows us to manipulate gene expression in Müller cells without affecting astrocytes in the retina. TRANSLATIONAL RELEVANCE: Gene manipulation under control of Mlc1 promoter offers novel technique to investigate the role of Müller cells.
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spelling pubmed-87642122022-01-26 The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina Danjo, Yosuke Shinozaki, Youichi Natsubori, Akiyo Kubota, Yuto Kashiwagi, Kenji Tanaka, Kenji F. Koizumi, Schuichi Transl Vis Sci Technol Article PURPOSE: Because the importance of glia in regulating brain functions has been demonstrated, genetic technologies that manipulate glial cell-specific gene expression in the brain have become essential and have made great progress. However, it is unknown whether the same strategy that is used in the brain can be applied to the retina because retinal glia differs from glia in the brain. Here, we aimed to find a method for selective gene expression in Müller cells (characteristic glial cells in the retina) and identified Mlc1 as a specific promoter of Müller cells. METHODS: Mlc1-tTA::Yellow-Cameleon-Nano(tetO/tetO) (YC-Nano) mice were used as a reporter line. YC-Nano, a fluorescent protein, was ectopically expressed in the cell type controlled by the Mlc1 promotor. Immunofluorescence staining was used to identify the cell type expressing YC-Nano protein. RESULTS: YC-Nano-positive ((+)) signals were observed as vertical stalks in the sliced retina and spanned from the nerve fiber layer through the outer nuclear layer. The density of YC-Nano(+) cells was higher around the optic nerve head and lower in the peripheral retina. The YC-Nano(+) signals colocalized with vimentin, a marker of Müller cells, but not with the cell markers for blood vessels, microglia, neurons, or astrocytes. CONCLUSIONS: The Mlc1 promoter allows us to manipulate gene expression in Müller cells without affecting astrocytes in the retina. TRANSLATIONAL RELEVANCE: Gene manipulation under control of Mlc1 promoter offers novel technique to investigate the role of Müller cells. The Association for Research in Vision and Ophthalmology 2022-01-18 /pmc/articles/PMC8764212/ /pubmed/35040915 http://dx.doi.org/10.1167/tvst.11.1.25 Text en Copyright 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
spellingShingle Article
Danjo, Yosuke
Shinozaki, Youichi
Natsubori, Akiyo
Kubota, Yuto
Kashiwagi, Kenji
Tanaka, Kenji F.
Koizumi, Schuichi
The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title_full The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title_fullStr The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title_full_unstemmed The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title_short The Mlc1 Promoter Directs Müller Cell-specific Gene Expression in the Retina
title_sort mlc1 promoter directs müller cell-specific gene expression in the retina
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8764212/
https://www.ncbi.nlm.nih.gov/pubmed/35040915
http://dx.doi.org/10.1167/tvst.11.1.25
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