Abstract
ABSTRACTGenetic, biochemical and histological studies have identified a number of different proteins as key drivers of human neurodegenerative diseases. Although different proteins are typically involved in different diseases, there is also considerable overlap. Addressing disease protein dysfunction in an in vivo neuronal context is often time consuming and requires labor-intensive analysis of transgenic models. To facilitate the rapid, cellular analysis of disease protein dysfunction, we have developed a fruit fly (Drosophila melanogaster) adult leg neuron assay. We tested the robustness of 41 transgenic fluorescent reporters and identified a number that were readily detected in the legs and could report on different cellular events. To test these reporters, we expressed a number of human proteins involved in neurodegenerative disease, in both their mutated and wild-type versions, to address the effects on reporter expression and localization. We observed strikingly different effects of the different disease proteins upon the various reporters with, for example, Aβ1-42 being highly neurotoxic, tau, parkin and HTT128Q affecting mitochondrial distribution, integrity or both, and Aβ1-42, tau, HTT128Q and ATX182Q affecting the F-actin network. This study provides proof of concept for using the Drosophila adult leg for inexpensive and rapid analysis of cellular effects of neurodegenerative disease proteins in mature neurons.
Highlights
Neurodegenerative diseases (NDs) have increasingly been linked to dysfunction of specific proteins, often unique to one disease, e.g. amyloid precursor protein (APP) to Alzheimer’s disease (AD), parkin (Park) to Parkinson’s disease (PD), huntingtin (HTT) to Huntington’s disease (HD), and superoxide dismutase (SOD1) to amyotrophic lateral sclerosis (ALS) (Kaur et al, 2016; Lill, 2016; Nopoulos, 2016; Selkoe and Hardy, 2016)
To expand the phenotypic read-out for protein neurotoxicity in vivo in Drosophila, we aimed to develop a method in which age-dependent analysis of neurotoxicity is possible, using fly leg neurons and axons
The Drosophila leg contains sensory neurons and their processes, in addition to the axonal processes and terminals from a number of leg motor neurons, all of which can be targeted by crossing UAS lines to the glutamatergic driver OK371-Gal4 (Baek and Mann, 2009)
Summary
Neurodegenerative diseases (NDs) have increasingly been linked to dysfunction of specific proteins, often unique to one disease, e.g. amyloid precursor protein (APP) to Alzheimer’s disease (AD), parkin (Park) to Parkinson’s disease (PD), huntingtin (HTT) to Huntington’s disease (HD), and superoxide dismutase (SOD1) to amyotrophic lateral sclerosis (ALS) (Kaur et al, 2016; Lill, 2016; Nopoulos, 2016; Selkoe and Hardy, 2016). Different ND proteins normally have distinct functions and subcellular locations, further supporting the notion of a certain degree of disease uniqueness. In contrast to this view of uniqueness, many ND. One possible reason for this dichotomy, at least in part, stems from the fact that it has been challenging to elucidate the in vivo role of the wild-type proteins and the dysfunction of the disease variants This is in part attributable to the slow progression of ND in mammalian model systems and to the difficulty with readily obtaining single-neuron cellular resolution in aging animals. The impact of ND proteins, normal or mutated, on different neuronal cellular events remains poorly understood
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