Related Experiment Video
Updated: Apr 23, 2026
Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster
Published on: March 14, 2019
In vivo evidence of pathogenicity of VPS35 mutations in the Drosophila
Hua-shan Wang1, Joanne Toh2, Patrick Ho3
1National Neuroscience Institute, 11 Jalan Tan Tock Seng, 308433, Singapore, Singapore. mitsuvolvo@gmail.com.
Abstract:
Mutations of VPS35, a component of the retromer complex have been associated with late onset familial Parkinson's disease. The D620N mutation in VPS35 appears to be most prevalent, however, P316S was found in two cases within the same family and a control, whereas L774M was identified in 6 cases and 1 control. In vivo evidence of their pathogenicity is lacking. Here we investigated the in vivo effects of P316S, D620N and L774M using Drosophila as a model. We generated transgenic human VPS35-expressing mutations and demonstrated that VPS35 D620N transgenic flies led to late-onset loss of TH-positive DA neurons, poor mobility, shortened lifespans and increased sensitivity to rotenone, a PD-linked environmental toxin, with some of these phenotypes observed for P316S but not in L774M transgenic flies. We conclude that D620N and to a smaller extent P316S are associated with pathogenicity in PD.
Insights
Mutations in VPS35 (Vacuolar protein sorting 35) are linked to Parkinson's disease (PD). The D620N and P316S mutations show pathogenicity in vivo, impacting neuron loss and mobility in a Drosophila model.
Area of Science:
- Neuroscience
- Genetics
- Cell Biology
Background:
- Mutations in VPS35 (Vacuolar protein sorting 35), a retromer complex component, are implicated in late-onset familial Parkinson's disease (PD).
- Specific mutations like D620N, P316S, and L774M have been identified in PD patients, but in vivo evidence for their pathogenicity is limited.
Purpose of the Study:
- To investigate the in vivo pathogenicity of VPS35 mutations (P316S, D620N, L774M) using a Drosophila melanogaster model.
- To assess the impact of these mutations on dopaminergic neuron survival, motor function, lifespan, and environmental toxin sensitivity.
Main Methods:
- Generation of transgenic Drosophila expressing human VPS35 with specific mutations (P316S, D620N, L774M).
- Assessment of tyrosine hydroxylase (TH)-positive dopaminergic neuron loss.
- Evaluation of fly mobility, lifespan, and sensitivity to rotenone, a Parkinson's disease-linked toxin.
Main Results:
- VPS35 D620N transgenic flies exhibited late-onset loss of TH-positive dopaminergic neurons, impaired mobility, reduced lifespan, and increased rotenone sensitivity.
- VPS35 P316S transgenic flies showed some similar, though less pronounced, phenotypes.
- VPS35 L774M transgenic flies did not display significant pathogenic phenotypes in this model.
Conclusions:
- The D620N mutation in VPS35 is demonstrably pathogenic in vivo, contributing to Parkinson's disease-like phenotypes.
- The P316S mutation also shows partial pathogenicity, while L774M appears less associated with disease in this model.
