Development of a Reporter System for In Vivo Monitoring of γ-Secretase Activity in Drosophila

Mol Cells. 2017 Jan;40(1):73-81. doi: 10.14348/molcells.2017.2294. Epub 2017 Jan 26.

Abstract

The γ-secretase complex represents an evolutionarily conserved family of transmembrane aspartyl proteases that cleave numerous type-I membrane proteins, including the β-amyloid precursor protein (APP) and the receptor Notch. All known rare mutations in APP and the γ-secretase catalytic component, presenilin, which lead to increased amyloid βpeptide production, are responsible for early-onset familial Alzheimer's disease. β-amyloid protein precursor-like (APPL) is the Drosophila ortholog of human APP. Here, we created Notch- and APPL-based Drosophila reporter systems for in vivo monitoring of γ-secretase activity. Ectopic expression of the Notch- and APPL-based chimeric reporters in wings results in vein truncation phenotypes. Reporter-mediated vein truncation phenotypes are enhanced by the Notch gain-of-function allele and suppressed by RNAi-mediated knockdown of presenilin. Furthermore, we find that apoptosis partly contributes to the vein truncation phenotypes of the APPL-based reporter, but not to the vein truncation phenotypes of the Notch-based reporter. Taken together, these results suggest that both in vivo reporter systems provide a powerful genetic tool to identify genes that modulate γ-secretase activity and/or APPL metabolism.

Keywords: APPL; Alzheimer’s disease; Notch; presenilin; γ-secretase.

MeSH terms

  • Amyloid Precursor Protein Secretases / analysis*
  • Amyloid Precursor Protein Secretases / genetics
  • Amyloid Precursor Protein Secretases / metabolism*
  • Animals
  • Drosophila / enzymology*
  • Drosophila / genetics
  • Female
  • Immunohistochemistry
  • Male
  • Mutation
  • Receptors, Notch / metabolism
  • Signal Transduction

Substances

  • Receptors, Notch
  • Amyloid Precursor Protein Secretases