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Formation of α-synuclein Lewy neurite-like aggregates in axons impedes the transport of distinct endosomes.

Volpicelli-Daley LA, Gamble KL, Schultheiss CE, Riddle DM, West AB, Lee VM - Mol. Biol. Cell (2014)

Bottom Line: Ultrastructural analyses and live imaging demonstrate that α-syn accumulations do not cause a generalized defect in axonal transport; the inclusions do not fill the axonal cytoplasm, disrupt the microtubule cytoskeleton, or affect the transport of synaptophysin or mitochondria.In addition, the TrkB receptor-associated signaling molecule pERK5 accumulates in α-syn aggregate-bearing neurons.These early effects of α-syn accumulations may predict points of intervention in the neurodegenerative process.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Behavioral Neurobiology, University of Alabama, Birmingham, Birmingham, AL 35294 Department of Pathology and Laboratory Medicine, Institute on Aging, and Center for Neurodegenerative Disease Research, University of Pennsylvania School of Medicine, Philadelphia, PA, 19104 volpicel@uab.edu.

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Normal transport of YFP-Mito in neurons with α-syn aggregates. Primary hippocampal neurons were transfected with YFP-Mito, treated with PBS or PFFs, and imaged 7 d later. YFP-Mito, number of particles analyzed, 162 for PBS and 244 for PFF (10 axons, PBS; 9 axons, PFF). (A) Top, images from movies captured every 1 s for 3 min; scale bar, 10 μm. Kymographs shown below the images were generated as visual representations of distance traveled over time. (B) Of the mobile particles, the percentages of anterograde and retrograde particles were quantified. There was no significant difference between the PBS- and PFF-treated groups. There were no significant differences in the mean number of YFP-Mito particles per 50 μm of axonal membrane (C), number of pauses (D), or number of reversals (E). A Poisson regression on velocities binned with 10 cut points was not statistically significant between PBS and PFF groups for anterograde YFP-Mito velocities (Wald χ2 = 0.713, p = NS; F) or for retrograde synaptophysin-GFP velocities (Wald χ2 = 2.886, p = NS; G). (F, G) Right, median and interquartile ranges of the velocities of the mobile synaptophysin-GFP particles. The Mann–Whitney test did not produce significant differences for anterograde or retrograde velocities.
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Figure 4: Normal transport of YFP-Mito in neurons with α-syn aggregates. Primary hippocampal neurons were transfected with YFP-Mito, treated with PBS or PFFs, and imaged 7 d later. YFP-Mito, number of particles analyzed, 162 for PBS and 244 for PFF (10 axons, PBS; 9 axons, PFF). (A) Top, images from movies captured every 1 s for 3 min; scale bar, 10 μm. Kymographs shown below the images were generated as visual representations of distance traveled over time. (B) Of the mobile particles, the percentages of anterograde and retrograde particles were quantified. There was no significant difference between the PBS- and PFF-treated groups. There were no significant differences in the mean number of YFP-Mito particles per 50 μm of axonal membrane (C), number of pauses (D), or number of reversals (E). A Poisson regression on velocities binned with 10 cut points was not statistically significant between PBS and PFF groups for anterograde YFP-Mito velocities (Wald χ2 = 0.713, p = NS; F) or for retrograde synaptophysin-GFP velocities (Wald χ2 = 2.886, p = NS; G). (F, G) Right, median and interquartile ranges of the velocities of the mobile synaptophysin-GFP particles. The Mann–Whitney test did not produce significant differences for anterograde or retrograde velocities.

Mentions: In addition, despite the presence of axonal α-syn aggregates, analyses of transport of a larger organelle such as the mitochondria using yellow fluorescent protein (YFP)–Mito (Figure 4A) revealed no difference between PBS- and PFF-treated groups for percentage of mobile particles (Figure 4B), abundance of YFP-Mito puncta (Figure 4C), number of pauses (Figure 4D), or number of reversals (Figure 4E). In addition, the Poisson regression analysis revealed no significant differences in anterograde or retrograde YFP-Mito velocities (Figure 4, F and G). Thus the presence of α-syn aggregates did not affect transport of synaptophysin or mitochondria.


Formation of α-synuclein Lewy neurite-like aggregates in axons impedes the transport of distinct endosomes.

Volpicelli-Daley LA, Gamble KL, Schultheiss CE, Riddle DM, West AB, Lee VM - Mol. Biol. Cell (2014)

Normal transport of YFP-Mito in neurons with α-syn aggregates. Primary hippocampal neurons were transfected with YFP-Mito, treated with PBS or PFFs, and imaged 7 d later. YFP-Mito, number of particles analyzed, 162 for PBS and 244 for PFF (10 axons, PBS; 9 axons, PFF). (A) Top, images from movies captured every 1 s for 3 min; scale bar, 10 μm. Kymographs shown below the images were generated as visual representations of distance traveled over time. (B) Of the mobile particles, the percentages of anterograde and retrograde particles were quantified. There was no significant difference between the PBS- and PFF-treated groups. There were no significant differences in the mean number of YFP-Mito particles per 50 μm of axonal membrane (C), number of pauses (D), or number of reversals (E). A Poisson regression on velocities binned with 10 cut points was not statistically significant between PBS and PFF groups for anterograde YFP-Mito velocities (Wald χ2 = 0.713, p = NS; F) or for retrograde synaptophysin-GFP velocities (Wald χ2 = 2.886, p = NS; G). (F, G) Right, median and interquartile ranges of the velocities of the mobile synaptophysin-GFP particles. The Mann–Whitney test did not produce significant differences for anterograde or retrograde velocities.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 4: Normal transport of YFP-Mito in neurons with α-syn aggregates. Primary hippocampal neurons were transfected with YFP-Mito, treated with PBS or PFFs, and imaged 7 d later. YFP-Mito, number of particles analyzed, 162 for PBS and 244 for PFF (10 axons, PBS; 9 axons, PFF). (A) Top, images from movies captured every 1 s for 3 min; scale bar, 10 μm. Kymographs shown below the images were generated as visual representations of distance traveled over time. (B) Of the mobile particles, the percentages of anterograde and retrograde particles were quantified. There was no significant difference between the PBS- and PFF-treated groups. There were no significant differences in the mean number of YFP-Mito particles per 50 μm of axonal membrane (C), number of pauses (D), or number of reversals (E). A Poisson regression on velocities binned with 10 cut points was not statistically significant between PBS and PFF groups for anterograde YFP-Mito velocities (Wald χ2 = 0.713, p = NS; F) or for retrograde synaptophysin-GFP velocities (Wald χ2 = 2.886, p = NS; G). (F, G) Right, median and interquartile ranges of the velocities of the mobile synaptophysin-GFP particles. The Mann–Whitney test did not produce significant differences for anterograde or retrograde velocities.
Mentions: In addition, despite the presence of axonal α-syn aggregates, analyses of transport of a larger organelle such as the mitochondria using yellow fluorescent protein (YFP)–Mito (Figure 4A) revealed no difference between PBS- and PFF-treated groups for percentage of mobile particles (Figure 4B), abundance of YFP-Mito puncta (Figure 4C), number of pauses (Figure 4D), or number of reversals (Figure 4E). In addition, the Poisson regression analysis revealed no significant differences in anterograde or retrograde YFP-Mito velocities (Figure 4, F and G). Thus the presence of α-syn aggregates did not affect transport of synaptophysin or mitochondria.

Bottom Line: Ultrastructural analyses and live imaging demonstrate that α-syn accumulations do not cause a generalized defect in axonal transport; the inclusions do not fill the axonal cytoplasm, disrupt the microtubule cytoskeleton, or affect the transport of synaptophysin or mitochondria.In addition, the TrkB receptor-associated signaling molecule pERK5 accumulates in α-syn aggregate-bearing neurons.These early effects of α-syn accumulations may predict points of intervention in the neurodegenerative process.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology and Behavioral Neurobiology, University of Alabama, Birmingham, Birmingham, AL 35294 Department of Pathology and Laboratory Medicine, Institute on Aging, and Center for Neurodegenerative Disease Research, University of Pennsylvania School of Medicine, Philadelphia, PA, 19104 volpicel@uab.edu.

Show MeSH
Related in: MedlinePlus