ZFIN ID: ZDB-PUB-190227-20
The epg5 knockout zebrafish line: a model to study Vici syndrome
Meneghetti, G., Skobo, T., Chrisam, M., Facchinello, N., Fontana, C.M., Bellesso, S., Sabatelli, P., Raggi, F., Cecconi, F., Bonaldo, P., Dalla Valle, L.
Date: 2019
Source: Autophagy   15(8): 1438-1454 (Journal)
Registered Authors: Bellesso, Stefania, Dalla Valle, Luisa, Facchinello, Nicola, Skobo, Tatjana
Keywords: , CRISPR-Cas9, Vici syndrome, autophagic flux, autophagy, zebrafish mutant line
MeSH Terms:
  • Agenesis of Corpus Callosum/metabolism*
  • Amino Acid Sequence
  • Animals
  • Autophagosomes/metabolism
  • Base Sequence
  • Cataract/metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Knockout Techniques*
  • Goblet Cells/pathology
  • Intestines/pathology
  • Intestines/ultrastructure
  • Larva/ultrastructure
  • Lysosomes/metabolism
  • Membrane Fusion
  • Models, Biological
  • Motor Neurons/metabolism
  • Motor Neurons/pathology
  • Mutagenesis/genetics
  • Mutation/genetics
  • Organ Specificity
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish Proteins/chemistry
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed: 30806141 Full text @ Autophagy
The EPG5 protein is a RAB7A effector involved in fusion specificity between autophagosomes and late endosomes or lysosomes during macroautophagy/autophagy. Mutations in the human EPG5 gene cause a rare and severe multisystem disorder called Vici syndrome. In this work, we show that zebrafish epg5-/- mutants from both heterozygous and incrossed homozygous matings are viable and can develop to the age of sexual maturity without conspicuous defects in external appearance. In agreement with the dysfunctional autophagy of Vici syndrome, western blot revealed higher levels of the Lc3-II autophagy marker in epg5-/- mutants with respect to wild type controls. Moreover, starvation elicited higher accumulation of Lc3-II in epg5-/- than in wild type larvae, together with a significant reduction of skeletal muscle birefringence. Accordingly, muscle ultrastructural analysis revealed accumulation of degradation-defective autolysosomes in starved epg5-/- mutants. By aging, epg5-/- mutants showed impaired motility and muscle thinning, together with accumulation of non-degradative autophagic vacuoles. Furthermore, epg5-/- adults displayed morphological alterations in gonads and heart. These findings point at the zebrafish epg5 mutant as a valuable model for EPG5-related disorders, thus providing a new tool for dissecting the contribution of EPG5 on the onset and progression of Vici syndrome as well as for the screening of autophagy-stimulating drugs.