ZFIN ID: ZDB-PUB-140613-2
Zebrafish ambra1a and ambra1b Knockdown Impairs Skeletal Muscle Development
Skobo, T., Benato, F., Grumati, P., Meneghetti, G., Cianfanelli, V., Castagnaro, S., Chrisam, M., Di Bartolomeo, S., Bonaldo, P., Cecconi, F., Valle, L.D.
Date: 2014
Source: PLoS One   9: e99210 (Journal)
Registered Authors: Benato, Francesca, Skobo, Tatjana
Keywords: Embryos, Skeletal muscles, Zebrafish, Muscle fibers, Autophagic cell death, Muscle proteins, Somites, Muscle differentiation
MeSH Terms:
  • Adaptor Proteins, Signal Transducing/deficiency
  • Adaptor Proteins, Signal Transducing/genetics
  • Adaptor Proteins, Signal Transducing/metabolism*
  • Animals
  • Birefringence
  • Cell Proliferation
  • Embryo, Nonmammalian/abnormalities
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/pathology
  • Gene Expression Regulation, Developmental/drug effects
  • Gene Knockdown Techniques*
  • Mice
  • Morpholinos/pharmacology
  • Movement
  • Muscle Development/genetics*
  • Muscle Fibers, Skeletal/drug effects
  • Muscle Fibers, Skeletal/pathology
  • Muscle Fibers, Skeletal/ultrastructure
  • Muscle, Skeletal/abnormalities
  • Muscle, Skeletal/drug effects
  • Muscle, Skeletal/embryology*
  • Muscle, Skeletal/metabolism
  • MyoD Protein/metabolism
  • Myosins/metabolism
  • PAX7 Transcription Factor/metabolism
  • Zebrafish/embryology*
  • Zebrafish/genetics*
  • Zebrafish Proteins/deficiency
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed: 24922546 Full text @ PLoS One

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.