Characterization of axonal transport defects in Drosophila Huntingtin mutants.
Polyglutamine (polyQ) expansion within Huntingtin (Htt) causes the fatal neurodegenerative disorder Huntington's Disease (HD). Although Htt is ubiquitously expressed and conserved from Drosophila to humans, its normal biological function is still being elucidated. Here we characterize a role for the Drosophila Htt homolog (dHtt) in fast axonal transport (FAT). Generation and expression of transgenic dHtt-mRFP and human Htt-mRFP fusion proteins in Drosophila revealed co-localization with mitochondria and synaptic vesicles undergoing FAT. However, Htt was not ubiquitously associated with the transport machinery, as it was excluded from dense-core vesicles and APLIP1 containing vesicles. Quantification of cargo movement in dHtt deficient axons revealed that mitochondria and synaptic vesicles show a decrease in the distance and duration of transport, and an increase in the number of pauses. In addition, the ratio of retrograde to anterograde flux was increased in mutant animals. Dense-core vesicles did not display similar defects in processivity, but did show altered retrograde to anterograde flux along axons. Given the co-localization with mitochondria and synaptic vesicles, but not dense-core vesicles, the data suggest dHtt likely acts locally at cargo interaction sites to regulate processivity. An increase in dynein heavy chain expression was also observed in dHtt mutants, suggesting that the altered flux observed for all cargo may represent secondary transport changes occurring independent of dHtt's primary function. Expression of dHtt in a milton (HAP1) mutant background revealed that the protein does not require mitochondria or HAP1 to localize along axons, suggesting Htt has an independent mechanism for coupling with motors to regulate their processivity during axonal transport.
a Department of Biology , The Picower Institute for Learning and Memory, Massachusetts Institute of Technology , Cambridge , MA , USA.|b Department of Brain and Cognitive Sciences , The Picower Institute for Learning and Memory, Massachusetts Institute of Technology , Cambridge , MA , USA.
Kurt R
KR
Weiss
a Department of Biology , The Picower Institute for Learning and Memory, Massachusetts Institute of Technology , Cambridge , MA , USA.|b Department of Brain and Cognitive Sciences , The Picower Institute for Learning and Memory, Massachusetts Institute of Technology , Cambridge , MA , USA.
J Troy
JT
Littleton
D000818:Animals; D030801:Animals, Genetically Modified; D001370:Axonal Transport / Q000235:genetics*; D004330:Drosophila; D006801:Humans; D000071058:Huntingtin Protein / Q000235:genetics*; D009154:Mutation
D000071058:Huntingtin Protein
Axonal transport; Drosophila; Huntington’s disease; neurodegeneration; polyglutamine