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Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis
BACKGROUND: Phosphoprotein signalling pathways have been intensively studied in vitro, yet their role in regulating tissue homeostasis is not fully understood. In the skin, interfollicular keratinocytes differentiate over approximately 2 weeks as they traverse the epidermis. The extracellular signal...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4678632/ https://www.ncbi.nlm.nih.gov/pubmed/26675891 http://dx.doi.org/10.1186/s13742-015-0102-5 |
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author | Cursons, Joseph Angel, Catherine E. Hurley, Daniel G. Print, Cristin G. Dunbar, P. Rod Jacobs, Marc D. Crampin, Edmund J. |
author_facet | Cursons, Joseph Angel, Catherine E. Hurley, Daniel G. Print, Cristin G. Dunbar, P. Rod Jacobs, Marc D. Crampin, Edmund J. |
author_sort | Cursons, Joseph |
collection | PubMed |
description | BACKGROUND: Phosphoprotein signalling pathways have been intensively studied in vitro, yet their role in regulating tissue homeostasis is not fully understood. In the skin, interfollicular keratinocytes differentiate over approximately 2 weeks as they traverse the epidermis. The extracellular signal-regulated kinase (ERK) branch of the mitogen-activated protein kinase (MAPK) pathway has been implicated in this process. Therefore, we examined ERK-MAPK activity within human epidermal keratinocytes in situ. FINDINGS: We used confocal microscopy and immunofluorescence labelling to measure the relative abundances of Raf-1, MEK1/2 and ERK1/2, and their phosphorylated (active) forms within three human skin samples. Additionally, we measured the abundance of selected proteins thought to modulate ERK-MAPK activity, including calmodulin, β1 integrin and stratifin (14-3-3σ); and of transcription factors known to act as effectors of ERK1/2, including the AP-1 components Jun-B, Fra2 and c-Fos. Imaging was performed with sufficient resolution to identify the plasma membrane, cytoplasm and nucleus as distinct domains within cells across the epidermis. The image field of view was also sufficiently large to capture the entire epidermis in cross-section, and thus the full range of keratinocyte differentiation in a single observation. Image processing methods were developed to quantify image data for mathematical and statistical analysis. Here, we provide raw image data and processed outputs. CONCLUSIONS: These data indicate coordinated changes in ERK-MAPK signalling activity throughout the depth of the epidermis, with changes in relative phosphorylation-mediated signalling activity occurring along the gradient of cellular differentiation. We believe these data provide unique information about intracellular signalling as they are obtained from a homeostatic human tissue, and they might be useful for investigating intercellular heterogeneity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13742-015-0102-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4678632 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-46786322015-12-16 Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis Cursons, Joseph Angel, Catherine E. Hurley, Daniel G. Print, Cristin G. Dunbar, P. Rod Jacobs, Marc D. Crampin, Edmund J. Gigascience Data Note BACKGROUND: Phosphoprotein signalling pathways have been intensively studied in vitro, yet their role in regulating tissue homeostasis is not fully understood. In the skin, interfollicular keratinocytes differentiate over approximately 2 weeks as they traverse the epidermis. The extracellular signal-regulated kinase (ERK) branch of the mitogen-activated protein kinase (MAPK) pathway has been implicated in this process. Therefore, we examined ERK-MAPK activity within human epidermal keratinocytes in situ. FINDINGS: We used confocal microscopy and immunofluorescence labelling to measure the relative abundances of Raf-1, MEK1/2 and ERK1/2, and their phosphorylated (active) forms within three human skin samples. Additionally, we measured the abundance of selected proteins thought to modulate ERK-MAPK activity, including calmodulin, β1 integrin and stratifin (14-3-3σ); and of transcription factors known to act as effectors of ERK1/2, including the AP-1 components Jun-B, Fra2 and c-Fos. Imaging was performed with sufficient resolution to identify the plasma membrane, cytoplasm and nucleus as distinct domains within cells across the epidermis. The image field of view was also sufficiently large to capture the entire epidermis in cross-section, and thus the full range of keratinocyte differentiation in a single observation. Image processing methods were developed to quantify image data for mathematical and statistical analysis. Here, we provide raw image data and processed outputs. CONCLUSIONS: These data indicate coordinated changes in ERK-MAPK signalling activity throughout the depth of the epidermis, with changes in relative phosphorylation-mediated signalling activity occurring along the gradient of cellular differentiation. We believe these data provide unique information about intracellular signalling as they are obtained from a homeostatic human tissue, and they might be useful for investigating intercellular heterogeneity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13742-015-0102-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-12-14 /pmc/articles/PMC4678632/ /pubmed/26675891 http://dx.doi.org/10.1186/s13742-015-0102-5 Text en © Cursons et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Data Note Cursons, Joseph Angel, Catherine E. Hurley, Daniel G. Print, Cristin G. Dunbar, P. Rod Jacobs, Marc D. Crampin, Edmund J. Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title | Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title_full | Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title_fullStr | Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title_full_unstemmed | Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title_short | Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis |
title_sort | spatially transformed fluorescence image data for erk-mapk and selected proteins within human epidermis |
topic | Data Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4678632/ https://www.ncbi.nlm.nih.gov/pubmed/26675891 http://dx.doi.org/10.1186/s13742-015-0102-5 |
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