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Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina

Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. However, the molecu...

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Autores principales: Todorova, Vyara, Merolla, Luca, Karademir, Duygu, Wögenstein, Gabriele M., Behr, Julian, Ebner, Lynn J. A., Samardzija, Marijana, Grimm, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9691741/
https://www.ncbi.nlm.nih.gov/pubmed/36424523
http://dx.doi.org/10.1038/s41598-022-24586-8
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author Todorova, Vyara
Merolla, Luca
Karademir, Duygu
Wögenstein, Gabriele M.
Behr, Julian
Ebner, Lynn J. A.
Samardzija, Marijana
Grimm, Christian
author_facet Todorova, Vyara
Merolla, Luca
Karademir, Duygu
Wögenstein, Gabriele M.
Behr, Julian
Ebner, Lynn J. A.
Samardzija, Marijana
Grimm, Christian
author_sort Todorova, Vyara
collection PubMed
description Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. However, the molecular analysis of photoreceptors is a challenge due to the heterogeneity of the retinal tissue and the lack of easy and reliable methods for cell separation. Here we present the ReLayS method—a simple technique for the separation of photoreceptor segments (PS) containing both inner and outer segments, outer nuclear layer (ONL), and inner retina (InR) that contains the remaining retinal layers. The layer-specific material isolated from a mouse half-retina with the ReLayS method was sufficient for protein isolation and Western blotting or RNA isolation and real-time PCR studies. The separation of PS, ONL, and InR was successfully validated by Western blotting and real-time PCR using proteins and genes with known expression profiles within the retina. Furthermore, the separation of the PS from the ONL enabled the detection of light-driven translocation of transducin from the PS to the soma. ReLayS is a simple and useful method to address protein and possibly metabolites distribution in photoreceptor compartments in various situations including development, ageing, and degenerative diseases.
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spelling pubmed-96917412022-11-26 Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina Todorova, Vyara Merolla, Luca Karademir, Duygu Wögenstein, Gabriele M. Behr, Julian Ebner, Lynn J. A. Samardzija, Marijana Grimm, Christian Sci Rep Article Understanding the physiology of the retina, and especially of the highly polarized photoreceptors, is essential not only to broaden our knowledge of the processes required for normal vision, but also to develop effective therapies to prevent or slow retinal degenerative diseases. However, the molecular analysis of photoreceptors is a challenge due to the heterogeneity of the retinal tissue and the lack of easy and reliable methods for cell separation. Here we present the ReLayS method—a simple technique for the separation of photoreceptor segments (PS) containing both inner and outer segments, outer nuclear layer (ONL), and inner retina (InR) that contains the remaining retinal layers. The layer-specific material isolated from a mouse half-retina with the ReLayS method was sufficient for protein isolation and Western blotting or RNA isolation and real-time PCR studies. The separation of PS, ONL, and InR was successfully validated by Western blotting and real-time PCR using proteins and genes with known expression profiles within the retina. Furthermore, the separation of the PS from the ONL enabled the detection of light-driven translocation of transducin from the PS to the soma. ReLayS is a simple and useful method to address protein and possibly metabolites distribution in photoreceptor compartments in various situations including development, ageing, and degenerative diseases. Nature Publishing Group UK 2022-11-23 /pmc/articles/PMC9691741/ /pubmed/36424523 http://dx.doi.org/10.1038/s41598-022-24586-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Todorova, Vyara
Merolla, Luca
Karademir, Duygu
Wögenstein, Gabriele M.
Behr, Julian
Ebner, Lynn J. A.
Samardzija, Marijana
Grimm, Christian
Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title_full Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title_fullStr Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title_full_unstemmed Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title_short Retinal Layer Separation (ReLayS) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
title_sort retinal layer separation (relays) method enables the molecular analysis of photoreceptor segments and cell bodies, as well as the inner retina
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9691741/
https://www.ncbi.nlm.nih.gov/pubmed/36424523
http://dx.doi.org/10.1038/s41598-022-24586-8
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