Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues
Precise localization of proteins and mRNA in histological sections is necessary for evaluating spatial gene expression patterns. Here we report sensitive detection of the gene products in fish tissues by immunohistochemistry (IHC) and in situ hybridization (ISH) assays on sections of whole specimens...
Published in: | Journal of Histochemistry & Cytochemistry |
---|---|
Main Authors: | , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
SAGE Publications
2009
|
Subjects: | |
Online Access: | http://dx.doi.org/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/pdf/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/full-xml/10.1369/jhc.2009.953695 |
id |
crsagepubl:10.1369/jhc.2009.953695 |
---|---|
record_format |
openpolar |
spelling |
crsagepubl:10.1369/jhc.2009.953695 2024-09-09T19:30:43+00:00 Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues Torgersen, Jacob S. Takle, Harald Andersen, Øivind 2009 http://dx.doi.org/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/pdf/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/full-xml/10.1369/jhc.2009.953695 en eng SAGE Publications http://journals.sagepub.com/page/policies/text-and-data-mining-license Journal of Histochemistry & Cytochemistry volume 57, issue 9, page 825-830 ISSN 0022-1554 1551-5044 journal-article 2009 crsagepubl https://doi.org/10.1369/jhc.2009.953695 2024-06-17T04:23:23Z Precise localization of proteins and mRNA in histological sections is necessary for evaluating spatial gene expression patterns. Here we report sensitive detection of the gene products in fish tissues by immunohistochemistry (IHC) and in situ hybridization (ISH) assays on sections of whole specimens and vertebra embedded in methyl methacrylate (MMA) resin. This plastic resin favors easy preparation of various specimen types and enables preparation of large sections with well-preserved cell morphology. IHC analysis of the muscle regulatory factor MyoD in transverse sections of juvenile cod revealed MyoD-positive cells in the dorsolateral parts of the adaxial muscle. ISH revealed less spatially restricted signals of the bone morphogenic protein bmp4 in muscle and brain. To assess the applicability of ISH on sections of bony tissue, col1a1 and col2a1 expression was investigated in non-decalcified vertebra sections of Atlantic salmon. The former was identified in both chondrocytes and osteoblasts, whereas the latter was mostly evident in chondrocytes. We conclude that MMA resin offers easy preparation of large and problematic tissues and the possibility of carrying out both IHC and ISH analyses using standard protocols. Article in Journal/Newspaper Atlantic salmon SAGE Publications Journal of Histochemistry & Cytochemistry 57 9 825 830 |
institution |
Open Polar |
collection |
SAGE Publications |
op_collection_id |
crsagepubl |
language |
English |
description |
Precise localization of proteins and mRNA in histological sections is necessary for evaluating spatial gene expression patterns. Here we report sensitive detection of the gene products in fish tissues by immunohistochemistry (IHC) and in situ hybridization (ISH) assays on sections of whole specimens and vertebra embedded in methyl methacrylate (MMA) resin. This plastic resin favors easy preparation of various specimen types and enables preparation of large sections with well-preserved cell morphology. IHC analysis of the muscle regulatory factor MyoD in transverse sections of juvenile cod revealed MyoD-positive cells in the dorsolateral parts of the adaxial muscle. ISH revealed less spatially restricted signals of the bone morphogenic protein bmp4 in muscle and brain. To assess the applicability of ISH on sections of bony tissue, col1a1 and col2a1 expression was investigated in non-decalcified vertebra sections of Atlantic salmon. The former was identified in both chondrocytes and osteoblasts, whereas the latter was mostly evident in chondrocytes. We conclude that MMA resin offers easy preparation of large and problematic tissues and the possibility of carrying out both IHC and ISH analyses using standard protocols. |
format |
Article in Journal/Newspaper |
author |
Torgersen, Jacob S. Takle, Harald Andersen, Øivind |
spellingShingle |
Torgersen, Jacob S. Takle, Harald Andersen, Øivind Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
author_facet |
Torgersen, Jacob S. Takle, Harald Andersen, Øivind |
author_sort |
Torgersen, Jacob S. |
title |
Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
title_short |
Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
title_full |
Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
title_fullStr |
Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
title_full_unstemmed |
Localization of mRNAs and Proteins in Methyl Methacrylate-embedded Tissues |
title_sort |
localization of mrnas and proteins in methyl methacrylate-embedded tissues |
publisher |
SAGE Publications |
publishDate |
2009 |
url |
http://dx.doi.org/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/pdf/10.1369/jhc.2009.953695 http://journals.sagepub.com/doi/full-xml/10.1369/jhc.2009.953695 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Journal of Histochemistry & Cytochemistry volume 57, issue 9, page 825-830 ISSN 0022-1554 1551-5044 |
op_rights |
http://journals.sagepub.com/page/policies/text-and-data-mining-license |
op_doi |
https://doi.org/10.1369/jhc.2009.953695 |
container_title |
Journal of Histochemistry & Cytochemistry |
container_volume |
57 |
container_issue |
9 |
container_start_page |
825 |
op_container_end_page |
830 |
_version_ |
1809899693153452032 |