The Development of an in vivo gamma-Secretase Assay using Zebrafish Embryos
- Authors
- Wilson, L., and Lardelli, M.
- ID
- ZDB-PUB-130610-59
- Date
- 2013
- Source
- Journal of Alzheimer's disease : JAD 36(3): 521-34 (Journal)
- Registered Authors
- Lardelli, Michael
- Keywords
- Alzheimer's disease, amyloid-β protein precursor, assay, γ-secretase, morpholinos, presenilin, truncated protein
- MeSH Terms
-
- Amyloid Precursor Protein Secretases/analysis*
- Amyloid Precursor Protein Secretases/genetics
- Amyloid Precursor Protein Secretases/metabolism
- Animals
- Blotting, Western
- Embryo, Nonmammalian
- Zebrafish
- PubMed
- 23666177 Full text @ J. Alzheimers Dis.
Aberrant proteolytic processing of amyloid-β protein precursor by γ-secretase complexes may result in an imbalance between production and clearance of the Aβ proteolytic product and promote neuronal dysfunction and death. Presenilin proteins form the catalytic core of γ-secretase complexes. The zebrafish, Danio rerio, is a versatile vertebrate model for investigating the molecular bases of Alzheimer's disease pathology. It possesses genes orthologous to human PSEN1 and PSEN2, and AβPP (psen1, psen2 and the genes appa and appb that are duplicates of an ancestral AβPP orthologue). Currently there is no in vivo assay appropriate for directly monitoring γ-secretase activity. Here, we describe such an assay in which the level of a γ-secretase substrate (a modified form of Appa protein) is observed in zebrafish embryos by western immunoblotting relative to a co-expressed protein not subject to γ-secretase activity. We have used the assay to analyze the effects on γ-secretase activity of blocking translation of zebrafish psen1 and/or psen2.