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Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity

BACKGROUND: The blood brain barrier (BBB) preserves neuronal function in the central nervous system (CNS) by tightly controlling metabolite exchanges with the blood. In the eye, the retina is likewise protected by the blood-retina barrier (BRB) to maintain phototransduction. We showed that the secre...

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Autores principales: Furtado, Jessica, Geraldo, Luiz Henrique, Leser, Felipe Saceanu, Poulet, Mathilde, Park, Hyojin, Pibouin-Fragner, Laurence, Eichmann, Anne, Boyé, Kevin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882365/
https://www.ncbi.nlm.nih.gov/pubmed/36711611
http://dx.doi.org/10.1101/2023.01.21.525006
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author Furtado, Jessica
Geraldo, Luiz Henrique
Leser, Felipe Saceanu
Poulet, Mathilde
Park, Hyojin
Pibouin-Fragner, Laurence
Eichmann, Anne
Boyé, Kevin
author_facet Furtado, Jessica
Geraldo, Luiz Henrique
Leser, Felipe Saceanu
Poulet, Mathilde
Park, Hyojin
Pibouin-Fragner, Laurence
Eichmann, Anne
Boyé, Kevin
author_sort Furtado, Jessica
collection PubMed
description BACKGROUND: The blood brain barrier (BBB) preserves neuronal function in the central nervous system (CNS) by tightly controlling metabolite exchanges with the blood. In the eye, the retina is likewise protected by the blood-retina barrier (BRB) to maintain phototransduction. We showed that the secreted guidance cue Netrin-1 regulated BBB integrity, by binding to endothelial Unc5B and regulating canonical β-catenin dependent expression of BBB gene expression OBJECTIVE: Here, we investigated if Netrin-1-binding to endothelial Unc5B also controlled BRB integrity, and if this process involved Norrin/β-catenin signaling, which is the major known driver of BRB development and maintenance. METHODS: We analyzed Tamoxifen-inducible loss- and gain- of-function alleles of Unc5B, Ntn1 and Ctnnb1 in conjunction with tracer injections and biochemical signaling studies. RESULTS: Inducible endothelial Unc5B deletion, and inducible global Ntn1 deletion in postnatal mice reduced phosphorylation of the Norrin receptor LRP5, leading to reduced β-catenin and LEF1 expression, conversion of retina endothelial cells from a barrier-competent Claudin-5+/PLVAP− state to a Claudin-5−/PLVAP+ leaky phenotype, and extravasation of injected low molecular weight tracers. Inducible Ctnnb1 gain of function rescued vascular leak in Unc5B mutants, and Ntn1 overexpression induced BRB tightening. Unc5B expression in pericytes contributed to BRB permeability, via regulation of endothelial Unc5B. Mechanistically, Netrin-1-Unc5B signaling promoted β-catenin dependent BRB signaling by enhancing phosphorylation of the Norrin receptor LRP5 via the Discs large homologue 1 (Dlg1) intracellular scaffolding protein. CONCLUSIONS: The data identify Netrin1-Unc5B as novel regulators of BRB integrity, with implications for diseases associated with BRB disruption.
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spelling pubmed-98823652023-01-28 Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity Furtado, Jessica Geraldo, Luiz Henrique Leser, Felipe Saceanu Poulet, Mathilde Park, Hyojin Pibouin-Fragner, Laurence Eichmann, Anne Boyé, Kevin bioRxiv Article BACKGROUND: The blood brain barrier (BBB) preserves neuronal function in the central nervous system (CNS) by tightly controlling metabolite exchanges with the blood. In the eye, the retina is likewise protected by the blood-retina barrier (BRB) to maintain phototransduction. We showed that the secreted guidance cue Netrin-1 regulated BBB integrity, by binding to endothelial Unc5B and regulating canonical β-catenin dependent expression of BBB gene expression OBJECTIVE: Here, we investigated if Netrin-1-binding to endothelial Unc5B also controlled BRB integrity, and if this process involved Norrin/β-catenin signaling, which is the major known driver of BRB development and maintenance. METHODS: We analyzed Tamoxifen-inducible loss- and gain- of-function alleles of Unc5B, Ntn1 and Ctnnb1 in conjunction with tracer injections and biochemical signaling studies. RESULTS: Inducible endothelial Unc5B deletion, and inducible global Ntn1 deletion in postnatal mice reduced phosphorylation of the Norrin receptor LRP5, leading to reduced β-catenin and LEF1 expression, conversion of retina endothelial cells from a barrier-competent Claudin-5+/PLVAP− state to a Claudin-5−/PLVAP+ leaky phenotype, and extravasation of injected low molecular weight tracers. Inducible Ctnnb1 gain of function rescued vascular leak in Unc5B mutants, and Ntn1 overexpression induced BRB tightening. Unc5B expression in pericytes contributed to BRB permeability, via regulation of endothelial Unc5B. Mechanistically, Netrin-1-Unc5B signaling promoted β-catenin dependent BRB signaling by enhancing phosphorylation of the Norrin receptor LRP5 via the Discs large homologue 1 (Dlg1) intracellular scaffolding protein. CONCLUSIONS: The data identify Netrin1-Unc5B as novel regulators of BRB integrity, with implications for diseases associated with BRB disruption. Cold Spring Harbor Laboratory 2023-01-21 /pmc/articles/PMC9882365/ /pubmed/36711611 http://dx.doi.org/10.1101/2023.01.21.525006 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Furtado, Jessica
Geraldo, Luiz Henrique
Leser, Felipe Saceanu
Poulet, Mathilde
Park, Hyojin
Pibouin-Fragner, Laurence
Eichmann, Anne
Boyé, Kevin
Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title_full Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title_fullStr Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title_full_unstemmed Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title_short Netrin-1 binding to Unc5B regulates Blood-Retina Barrier integrity
title_sort netrin-1 binding to unc5b regulates blood-retina barrier integrity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9882365/
https://www.ncbi.nlm.nih.gov/pubmed/36711611
http://dx.doi.org/10.1101/2023.01.21.525006
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