The Development of an in vivo γ-Secretase Assay using Zebrafish Embryos

被引：8

作者：

Wilson, Lachlan ^{[1
]}

Lardelli, Michael ^{[1
]}

机构：

[1] Univ Adelaide, Sch Mol & Biomed Sci, Discipline Genet, Adelaide, SA 5005, Australia

来源：

JOURNAL OF ALZHEIMERS DISEASE | 2013年 / 36卷 / 03期

基金：

英国医学研究理事会;

关键词：

Alzheimer's disease; amyloid-beta protein precursor; assay; gamma-secretase; morpholinos; presenilin; truncated protein; zebrafish; AMYLOID PRECURSOR PROTEIN; FAMILIAL ALZHEIMERS-DISEASE; CAPACITATIVE CALCIUM-ENTRY; INTRACELLULAR DOMAIN AICD; BETA-PROTEIN; INTRAMEMBRANE CLEAVAGE; TRANSMEMBRANE DOMAIN; SIGNAL-TRANSDUCTION; MUTANT PRESENILIN-1; TERMINAL FRAGMENT;

D O I：

10.3233/JAD-130332

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

Aberrant proteolytic processing of amyloid-beta protein precursor by gamma-secretase complexes may result in an imbalance between production and clearance of the A beta proteolytic product and promote neuronal dysfunction and death. Presenilin proteins form the catalytic core of gamma-secretase complexes. The zebrafish, Danio rerio, is a versatile vertebrate model for investigating the molecular basis of Alzheimer's disease pathology. It possesses genes orthologous to human PSEN1 and PSEN2 (psen1 and psen2 respectively), and A beta PP (appa and appb that are duplicates of an ancestral A beta PP orthologue). Currently there is no in vivo assay appropriate for directly monitoring gamma-secretase activity. Here, we describe such an assay in which the level of a gamma-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 gamma-secretase activity. We have used the assay to analyze the effects on gamma-secretase activity of blocking translation of transcripts of zebrafish psen1 and/or psen2.

引用

页码：521 / 534

页数：14

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