Expression pattern and functions of autophagy-related gene atg5 in zebrafish organogenesis

Autophagy. 2011 Dec;7(12):1514-27. doi: 10.4161/auto.7.12.18040.

Abstract

The implications of autophagy-related genes in serious neural degenerative diseases have been well documented. However, the functions and regulation of the family genes in embryonic development remain to be rigorously studied. Here, we report on for the first time the important role of atg5 gene in zebrafish neurogenesis and organogenesis as evidenced by the spatiotemporal expression pattern and functional analysis. Using morpholino oligo knockdown and mRNA overexpression, we demonstrated that zebrafish atg5 is required for normal morphogenesis of brain regionalization and body plan as well as for expression regulation of neural gene markers: gli1, huC, nkx2.2, pink1, β-synuclein, xb51 and zic1. We further demonstrated that ATG5 protein is involved in autophagy by LC3-II/LC3I ratio and rapamycin-induction experiments, and that ATG5 is capable of regulating expression of itself gene in the manner of a feedback inhibition loop. In addition, we found that expression of another autophagy-related gene, atg12, is maintained at a higher constant level like a housekeeping gene. This indicates that the formation of the ATG12–ATG5 conjugate may be dependent on ATG5 protein generation and its splicing, rather than on ATG12 protein in zebrafish. Importantly, in the present study, we provide a mechanistic insight into the regulation and functional roles of atg5 in development of zebrafish nervous system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autophagy / drug effects
  • Autophagy / genetics*
  • Autophagy-Related Protein 5
  • Embryo, Nonmammalian / abnormalities
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / metabolism
  • Embryonic Development / drug effects
  • Embryonic Development / genetics
  • Feedback, Physiological / drug effects
  • Gene Expression Regulation, Developmental* / drug effects
  • Models, Biological
  • Molecular Sequence Data
  • Organogenesis / drug effects
  • Organogenesis / genetics*
  • Sequence Homology, Amino Acid
  • Sirolimus / pharmacology
  • Time Factors
  • Transcription, Genetic / drug effects
  • Zebrafish / embryology*
  • Zebrafish / genetics*
  • Zebrafish Proteins / chemistry
  • Zebrafish Proteins / genetics*
  • Zebrafish Proteins / metabolism

Substances

  • Atg5 protein, zebrafish
  • Autophagy-Related Protein 5
  • Zebrafish Proteins
  • Sirolimus