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Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells

Retina formation requires the correct spatiotemporal patterning of key regulatory factors. While it is known that repression of several signaling pathways lead to specification of retinal fates, addition of only Noggin, a known BMP antagonist, can convert pluripotent Xenopus laevis animal cap cells...

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Autores principales: Wong, Kimberly A., Trembley, Michael, Abd Wahab, Syafiq, Viczian, Andrea S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4400599/
https://www.ncbi.nlm.nih.gov/pubmed/25750435
http://dx.doi.org/10.1242/bio.20149977
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author Wong, Kimberly A.
Trembley, Michael
Abd Wahab, Syafiq
Viczian, Andrea S.
author_facet Wong, Kimberly A.
Trembley, Michael
Abd Wahab, Syafiq
Viczian, Andrea S.
author_sort Wong, Kimberly A.
collection PubMed
description Retina formation requires the correct spatiotemporal patterning of key regulatory factors. While it is known that repression of several signaling pathways lead to specification of retinal fates, addition of only Noggin, a known BMP antagonist, can convert pluripotent Xenopus laevis animal cap cells to functional retinal cells. The aim of this study is to determine the intracellular molecular events that occur during this conversion. Surprisingly, blocking BMP signaling alone failed to mimic Noggin treatment. Overexpressing Noggin in pluripotent cells resulted in a concentration-dependent suppression of both Smad1 and Smad2 phosphorylation, which act downstream of BMP and Activin signaling, respectively. This caused a decrease in downstream targets: endothelial marker, xk81, and mesodermal marker, xbra. We treated pluripotent cells with dominant-negative receptors or the chemical inhibitors, dorsomorphin and SB431542, which each target either the BMP or Activin signaling pathway. We determined the effect of these treatments on retina formation using the Animal Cap Transplant (ACT) assay; in which treated pluripotent cells were transplanted into the eye field of host embryos. We found that inhibition of Activin signaling, in the presence of BMP signaling inhibition, promotes efficient retinal specification in Xenopus tissue, mimicking the affect of adding Noggin alone. In whole embryos, we found that the eye field marker, rax, expanded when adding both dominant-negative Smad1 and Smad2, as did treating the cells with both dorsomorphin and SB431542. Future studies could translate these findings to a mammalian culture assay, in order to more efficiently produce retinal cells in culture.
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spelling pubmed-44005992015-04-24 Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells Wong, Kimberly A. Trembley, Michael Abd Wahab, Syafiq Viczian, Andrea S. Biol Open Research Article Retina formation requires the correct spatiotemporal patterning of key regulatory factors. While it is known that repression of several signaling pathways lead to specification of retinal fates, addition of only Noggin, a known BMP antagonist, can convert pluripotent Xenopus laevis animal cap cells to functional retinal cells. The aim of this study is to determine the intracellular molecular events that occur during this conversion. Surprisingly, blocking BMP signaling alone failed to mimic Noggin treatment. Overexpressing Noggin in pluripotent cells resulted in a concentration-dependent suppression of both Smad1 and Smad2 phosphorylation, which act downstream of BMP and Activin signaling, respectively. This caused a decrease in downstream targets: endothelial marker, xk81, and mesodermal marker, xbra. We treated pluripotent cells with dominant-negative receptors or the chemical inhibitors, dorsomorphin and SB431542, which each target either the BMP or Activin signaling pathway. We determined the effect of these treatments on retina formation using the Animal Cap Transplant (ACT) assay; in which treated pluripotent cells were transplanted into the eye field of host embryos. We found that inhibition of Activin signaling, in the presence of BMP signaling inhibition, promotes efficient retinal specification in Xenopus tissue, mimicking the affect of adding Noggin alone. In whole embryos, we found that the eye field marker, rax, expanded when adding both dominant-negative Smad1 and Smad2, as did treating the cells with both dorsomorphin and SB431542. Future studies could translate these findings to a mammalian culture assay, in order to more efficiently produce retinal cells in culture. The Company of Biologists 2015-03-06 /pmc/articles/PMC4400599/ /pubmed/25750435 http://dx.doi.org/10.1242/bio.20149977 Text en © 2015. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Research Article
Wong, Kimberly A.
Trembley, Michael
Abd Wahab, Syafiq
Viczian, Andrea S.
Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title_full Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title_fullStr Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title_full_unstemmed Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title_short Efficient retina formation requires suppression of both Activin and BMP signaling pathways in pluripotent cells
title_sort efficient retina formation requires suppression of both activin and bmp signaling pathways in pluripotent cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4400599/
https://www.ncbi.nlm.nih.gov/pubmed/25750435
http://dx.doi.org/10.1242/bio.20149977
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