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Characterization of Hspb8 in Zebrafish

Hspb8 is a member of the small heat shock protein (sHSP) family. Its expression is known to be upregulated under heat shock. This protein interacts with different partners and can, therefore, be involved in various processes relevant to tissue integrity and functioning. In humans, mutations in the g...

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Autores principales: Dubińska-Magiera, Magda, Niedbalska-Tarnowska, Joanna, Migocka-Patrzałek, Marta, Posyniak, Ewelina, Daczewska, Małgorzata
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7348923/
https://www.ncbi.nlm.nih.gov/pubmed/32604890
http://dx.doi.org/10.3390/cells9061562
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author Dubińska-Magiera, Magda
Niedbalska-Tarnowska, Joanna
Migocka-Patrzałek, Marta
Posyniak, Ewelina
Daczewska, Małgorzata
author_facet Dubińska-Magiera, Magda
Niedbalska-Tarnowska, Joanna
Migocka-Patrzałek, Marta
Posyniak, Ewelina
Daczewska, Małgorzata
author_sort Dubińska-Magiera, Magda
collection PubMed
description Hspb8 is a member of the small heat shock protein (sHSP) family. Its expression is known to be upregulated under heat shock. This protein interacts with different partners and can, therefore, be involved in various processes relevant to tissue integrity and functioning. In humans, mutations in the gene encoding Hspb8 can lead to the development of various diseases such as myopathies and neuropathies. In our study, we aimed to perform an in-depth characterization of zebrafish Hspb8 during zebrafish development. We applied techniques such as RT-qPCR, Western blot, immunofluorescence, co-immunoprecipitation, LC-MS, and morpholino-mediated knockdown. We broadened the knowledge regarding zebrafish hspb8 expression during development under normal and heat shock conditions as well as its tissue- and subcellular-specific localization. A co-IP analysis allowed us to conclude that zebrafish Hspb8 can interact with proteins such as Bag3 and Hsc70, which are crucial for formation of an autophagy-inducing complex. We also demonstrated that hspb8 morpholino-mediated knockdown has an impact on zebrafish embryos’ morphology, muscle ultrastructure, and motility behavior. Our research provides a valuable resource for the potential use of the zebrafish as a model for studying pathological conditions associated with hspb8 disorders.
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spelling pubmed-73489232020-07-22 Characterization of Hspb8 in Zebrafish Dubińska-Magiera, Magda Niedbalska-Tarnowska, Joanna Migocka-Patrzałek, Marta Posyniak, Ewelina Daczewska, Małgorzata Cells Article Hspb8 is a member of the small heat shock protein (sHSP) family. Its expression is known to be upregulated under heat shock. This protein interacts with different partners and can, therefore, be involved in various processes relevant to tissue integrity and functioning. In humans, mutations in the gene encoding Hspb8 can lead to the development of various diseases such as myopathies and neuropathies. In our study, we aimed to perform an in-depth characterization of zebrafish Hspb8 during zebrafish development. We applied techniques such as RT-qPCR, Western blot, immunofluorescence, co-immunoprecipitation, LC-MS, and morpholino-mediated knockdown. We broadened the knowledge regarding zebrafish hspb8 expression during development under normal and heat shock conditions as well as its tissue- and subcellular-specific localization. A co-IP analysis allowed us to conclude that zebrafish Hspb8 can interact with proteins such as Bag3 and Hsc70, which are crucial for formation of an autophagy-inducing complex. We also demonstrated that hspb8 morpholino-mediated knockdown has an impact on zebrafish embryos’ morphology, muscle ultrastructure, and motility behavior. Our research provides a valuable resource for the potential use of the zebrafish as a model for studying pathological conditions associated with hspb8 disorders. MDPI 2020-06-26 /pmc/articles/PMC7348923/ /pubmed/32604890 http://dx.doi.org/10.3390/cells9061562 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dubińska-Magiera, Magda
Niedbalska-Tarnowska, Joanna
Migocka-Patrzałek, Marta
Posyniak, Ewelina
Daczewska, Małgorzata
Characterization of Hspb8 in Zebrafish
title Characterization of Hspb8 in Zebrafish
title_full Characterization of Hspb8 in Zebrafish
title_fullStr Characterization of Hspb8 in Zebrafish
title_full_unstemmed Characterization of Hspb8 in Zebrafish
title_short Characterization of Hspb8 in Zebrafish
title_sort characterization of hspb8 in zebrafish
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7348923/
https://www.ncbi.nlm.nih.gov/pubmed/32604890
http://dx.doi.org/10.3390/cells9061562
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