Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.

The essential role of autophagy in muscle homeostasis has been clearly demonstrated by phenotype analysis of mice with muscle-specific inactivation of genes encoding autophagy-related proteins. Ambra1 is a key component of the Beclin 1 complex and, in zebrafish, it is encoded by two paralogous genes...

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Main Authors: Tatjana Skobo, Francesca Benato, Paolo Grumati, Giacomo Meneghetti, Valentina Cianfanelli, Silvia Castagnaro, Martina Chrisam, Sabrina Di Bartolomeo, Paolo Bonaldo, Francesco Cecconi, Luisa Dalla Valle
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0099210
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author Tatjana Skobo
Francesca Benato
Paolo Grumati
Giacomo Meneghetti
Valentina Cianfanelli
Silvia Castagnaro
Martina Chrisam
Sabrina Di Bartolomeo
Paolo Bonaldo
Francesco Cecconi
Luisa Dalla Valle
author_facet Tatjana Skobo
Francesca Benato
Paolo Grumati
Giacomo Meneghetti
Valentina Cianfanelli
Silvia Castagnaro
Martina Chrisam
Sabrina Di Bartolomeo
Paolo Bonaldo
Francesco Cecconi
Luisa Dalla Valle
author_sort Tatjana Skobo
collection DOAJ
description The essential role of autophagy in muscle homeostasis has been clearly demonstrated by phenotype analysis of mice with muscle-specific inactivation of genes encoding autophagy-related proteins. Ambra1 is a key component of the Beclin 1 complex and, in zebrafish, it is encoded by two paralogous genes, ambra1a and ambra1b, both required for normal embryogenesis and larval development. In this study we focused on the function of Ambra1, a positive regulator of the autophagic process, during skeletal muscle development by means of morpholino (MO)-mediated knockdown and compared the phenotype of zebrafish Ambra1-depleted embryos with that of Ambra1gt/gt mouse embryos. Morphological analysis of zebrafish morphant embryos revealed that silencing of ambra1 impairs locomotor activity and muscle development, as well as myoD1 expression. Skeletal muscles in ATG-morphant embryos displayed severe histopathological changes and contained only small areas of organized myofibrils that were widely dispersed throughout the cell. Double knockdown of ambra1a and ambra1b resulted in a more severe phenotype whereas defects were much less evident in splice-morphants. The morphants phenotypes were effectively rescued by co-injection with human AMBRA1 mRNA. Together, these results indicate that ambra1a and ambra1b are required for the correct development and morphogenesis of skeletal muscle.
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spelling doaj-art-a663d098e1a04e87ab2747a2f631baad2025-08-20T02:34:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0196e9921010.1371/journal.pone.0099210Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.Tatjana SkoboFrancesca BenatoPaolo GrumatiGiacomo MeneghettiValentina CianfanelliSilvia CastagnaroMartina ChrisamSabrina Di BartolomeoPaolo BonaldoFrancesco CecconiLuisa Dalla ValleThe essential role of autophagy in muscle homeostasis has been clearly demonstrated by phenotype analysis of mice with muscle-specific inactivation of genes encoding autophagy-related proteins. Ambra1 is a key component of the Beclin 1 complex and, in zebrafish, it is encoded by two paralogous genes, ambra1a and ambra1b, both required for normal embryogenesis and larval development. In this study we focused on the function of Ambra1, a positive regulator of the autophagic process, during skeletal muscle development by means of morpholino (MO)-mediated knockdown and compared the phenotype of zebrafish Ambra1-depleted embryos with that of Ambra1gt/gt mouse embryos. Morphological analysis of zebrafish morphant embryos revealed that silencing of ambra1 impairs locomotor activity and muscle development, as well as myoD1 expression. Skeletal muscles in ATG-morphant embryos displayed severe histopathological changes and contained only small areas of organized myofibrils that were widely dispersed throughout the cell. Double knockdown of ambra1a and ambra1b resulted in a more severe phenotype whereas defects were much less evident in splice-morphants. The morphants phenotypes were effectively rescued by co-injection with human AMBRA1 mRNA. Together, these results indicate that ambra1a and ambra1b are required for the correct development and morphogenesis of skeletal muscle.https://doi.org/10.1371/journal.pone.0099210
spellingShingle Tatjana Skobo
Francesca Benato
Paolo Grumati
Giacomo Meneghetti
Valentina Cianfanelli
Silvia Castagnaro
Martina Chrisam
Sabrina Di Bartolomeo
Paolo Bonaldo
Francesco Cecconi
Luisa Dalla Valle
Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
PLoS ONE
title Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
title_full Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
title_fullStr Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
title_full_unstemmed Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
title_short Zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development.
title_sort zebrafish ambra1a and ambra1b knockdown impairs skeletal muscle development
url https://doi.org/10.1371/journal.pone.0099210
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