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Patched1 and Patched2 inhibit Smoothened non-cell autonomously

Smoothened (Smo) inhibition by Patched (Ptch) is central to Hedgehog (Hh) signaling. Ptch, a proton driven antiporter, is required for Smo inhibition via an unknown mechanism. Hh ligand binding to Ptch reverses this inhibition and activated Smo initiates the Hh response. To determine whether Ptch in...

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Autores principales: Roberts, Brock, Casillas, Catalina, Alfaro, Astrid C, Jägers, Carina, Roelink, Henk
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014547/
https://www.ncbi.nlm.nih.gov/pubmed/27552050
http://dx.doi.org/10.7554/eLife.17634
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author Roberts, Brock
Casillas, Catalina
Alfaro, Astrid C
Jägers, Carina
Roelink, Henk
author_facet Roberts, Brock
Casillas, Catalina
Alfaro, Astrid C
Jägers, Carina
Roelink, Henk
author_sort Roberts, Brock
collection PubMed
description Smoothened (Smo) inhibition by Patched (Ptch) is central to Hedgehog (Hh) signaling. Ptch, a proton driven antiporter, is required for Smo inhibition via an unknown mechanism. Hh ligand binding to Ptch reverses this inhibition and activated Smo initiates the Hh response. To determine whether Ptch inhibits Smo strictly in the same cell or also mediates non-cell-autonomous Smo inhibition, we generated genetically mosaic neuralized embryoid bodies (nEBs) from mouse embryonic stem cells (mESCs). These experiments utilized novel mESC lines in which Ptch1, Ptch2, Smo, Shh and 7dhcr were inactivated via gene editing in multiple combinations, allowing us to measure non-cell autonomous interactions between cells with differing Ptch1/2 status. In several independent assays, the Hh response was repressed by Ptch1/2 in nearby cells. When 7dhcr was targeted, cells displayed elevated non-cell autonomous inhibition. These findings support a model in which Ptch1/2 mediate secretion of a Smo-inhibitory cholesterol precursor. DOI: http://dx.doi.org/10.7554/eLife.17634.001
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spelling pubmed-50145472016-09-09 Patched1 and Patched2 inhibit Smoothened non-cell autonomously Roberts, Brock Casillas, Catalina Alfaro, Astrid C Jägers, Carina Roelink, Henk eLife Developmental Biology and Stem Cells Smoothened (Smo) inhibition by Patched (Ptch) is central to Hedgehog (Hh) signaling. Ptch, a proton driven antiporter, is required for Smo inhibition via an unknown mechanism. Hh ligand binding to Ptch reverses this inhibition and activated Smo initiates the Hh response. To determine whether Ptch inhibits Smo strictly in the same cell or also mediates non-cell-autonomous Smo inhibition, we generated genetically mosaic neuralized embryoid bodies (nEBs) from mouse embryonic stem cells (mESCs). These experiments utilized novel mESC lines in which Ptch1, Ptch2, Smo, Shh and 7dhcr were inactivated via gene editing in multiple combinations, allowing us to measure non-cell autonomous interactions between cells with differing Ptch1/2 status. In several independent assays, the Hh response was repressed by Ptch1/2 in nearby cells. When 7dhcr was targeted, cells displayed elevated non-cell autonomous inhibition. These findings support a model in which Ptch1/2 mediate secretion of a Smo-inhibitory cholesterol precursor. DOI: http://dx.doi.org/10.7554/eLife.17634.001 eLife Sciences Publications, Ltd 2016-08-23 /pmc/articles/PMC5014547/ /pubmed/27552050 http://dx.doi.org/10.7554/eLife.17634 Text en © 2016, Roberts et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Developmental Biology and Stem Cells
Roberts, Brock
Casillas, Catalina
Alfaro, Astrid C
Jägers, Carina
Roelink, Henk
Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title_full Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title_fullStr Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title_full_unstemmed Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title_short Patched1 and Patched2 inhibit Smoothened non-cell autonomously
title_sort patched1 and patched2 inhibit smoothened non-cell autonomously
topic Developmental Biology and Stem Cells
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014547/
https://www.ncbi.nlm.nih.gov/pubmed/27552050
http://dx.doi.org/10.7554/eLife.17634
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