Cargando…
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...
Autores principales: | , , , , , , |
---|---|
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 |
_version_ | 1784634114847014912 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8764212 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Association for Research in Vision and Ophthalmology |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT danjoyosuke themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT shinozakiyouichi themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT natsuboriakiyo themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT kubotayuto themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT kashiwagikenji themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT tanakakenjif themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT koizumischuichi themlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT danjoyosuke mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT shinozakiyouichi mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT natsuboriakiyo mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT kubotayuto mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT kashiwagikenji mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT tanakakenjif mlc1promoterdirectsmullercellspecificgeneexpressionintheretina AT koizumischuichi mlc1promoterdirectsmullercellspecificgeneexpressionintheretina |