Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction

Improved in situ hybridization methods for mRNA detection in tissues have been developed based on the hybridization chain reaction (HCR). We show that in situ HCR methods can be used for the detection of microRNAs in tissue sections from mouse retinas. In situ HCR can be used for the detection of tw...

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Autores principales: Zhuang, Pei, Zhang, Huanqing, Welchko, Ryan M., Thompson, Robert C., Xu, Shunbin, Turner, David L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6962165/
https://www.ncbi.nlm.nih.gov/pubmed/31942002
http://dx.doi.org/10.1038/s41598-019-57194-0
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author Zhuang, Pei
Zhang, Huanqing
Welchko, Ryan M.
Thompson, Robert C.
Xu, Shunbin
Turner, David L.
author_facet Zhuang, Pei
Zhang, Huanqing
Welchko, Ryan M.
Thompson, Robert C.
Xu, Shunbin
Turner, David L.
author_sort Zhuang, Pei
collection PubMed
description Improved in situ hybridization methods for mRNA detection in tissues have been developed based on the hybridization chain reaction (HCR). We show that in situ HCR methods can be used for the detection of microRNAs in tissue sections from mouse retinas. In situ HCR can be used for the detection of two microRNAs simultaneously or for the combined detection of microRNA and mRNA. In addition, miRNA in situ HCR can be combined with immunodetection of proteins. We use these methods to characterize cells expressing specific microRNAs in the mouse retina. We find that miR-181a is expressed in amacrine cells during development and in adult retinas, and it is present in both GABAergic and glycinergic amacrine cells. The detection of microRNAs with in situ HCR should facilitate studies of microRNA function and gene regulation in the retina and other tissues.
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spelling pubmed-69621652020-01-23 Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction Zhuang, Pei Zhang, Huanqing Welchko, Ryan M. Thompson, Robert C. Xu, Shunbin Turner, David L. Sci Rep Article Improved in situ hybridization methods for mRNA detection in tissues have been developed based on the hybridization chain reaction (HCR). We show that in situ HCR methods can be used for the detection of microRNAs in tissue sections from mouse retinas. In situ HCR can be used for the detection of two microRNAs simultaneously or for the combined detection of microRNA and mRNA. In addition, miRNA in situ HCR can be combined with immunodetection of proteins. We use these methods to characterize cells expressing specific microRNAs in the mouse retina. We find that miR-181a is expressed in amacrine cells during development and in adult retinas, and it is present in both GABAergic and glycinergic amacrine cells. The detection of microRNAs with in situ HCR should facilitate studies of microRNA function and gene regulation in the retina and other tissues. Nature Publishing Group UK 2020-01-15 /pmc/articles/PMC6962165/ /pubmed/31942002 http://dx.doi.org/10.1038/s41598-019-57194-0 Text en © The Author(s) 2020 Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Zhuang, Pei
Zhang, Huanqing
Welchko, Ryan M.
Thompson, Robert C.
Xu, Shunbin
Turner, David L.
Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title_full Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title_fullStr Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title_full_unstemmed Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title_short Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction
title_sort combined microrna and mrna detection in mammalian retinas by in situ hybridization chain reaction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6962165/
https://www.ncbi.nlm.nih.gov/pubmed/31942002
http://dx.doi.org/10.1038/s41598-019-57194-0
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