Synaptic vesicle glycoprotein 2C enhances vesicular storage of dopamine and counters dopaminergic toxicity
Data files
Apr 12, 2024 version files 14.87 MB
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Individual_folders.zip
13.83 MB
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README.md
2.31 KB
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SV2C_manuscript_data.pzfx
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Abstract
Dopaminergic neurons of the substantia nigra exist in a persistent state of vulnerability resulting from high baseline oxidative stress, high energy demand, and broad unmyelinated axonal arborizations. Impairments in the storage of dopamine compound this stress due to cytosolic reactions that transform the vital neurotransmitter into an endogenous neurotoxicant, and this toxicity is thought to contribute to the dopamine neuron degeneration that occurs Parkinson’s disease. We have previously identified synaptic vesicle glycoprotein 2C (SV2C) as a modifier of vesicular dopamine function, demonstrating that genetic ablation of SV2C in mice results in decreased dopamine content and evoked dopamine release in the striatum. Here, we adapted a previously published in vitro assay utilizing false fluorescent neurotransmitter 206 (FFN206) to visualize how SV2C regulates vesicular dopamine dynamics and identified that SV2C promotes the uptake and retention of FFN206 within vesicles. In addition, we present data indicating that SV2C enhances the retention of dopamine in the vesicular compartment with radiolabeled dopamine in vesicles isolated from immortalized cells and from mouse brain. Further, we demonstrate that SV2C enhances the ability of vesicles to store the neurotoxicant 1-methyl-4-phenylpyridinium (MPP+) and that genetic ablation of SV2C results in enhanced 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced vulnerability in mice. Together, these findings establish that SV2C functions to enhance storage of dopamine and toxicants and helps maintain the integrity of dopaminergic neurons.
README: Synaptic vesicle glycoprotein 2C enhances vesicular storage of dopamine and counters dopaminergic toxicity
This dataset contains the raw data corresponding to the manuscript Synaptic vesicle glycoprotein 2C enhances vesicular storage of dopamine and counters dopaminergic toxicity. Inclusive in this dataset is the following: 1) a GraphPad Prism file containing all of the data found in the manuscript with statistical analysis and graphs; 2) individual .csv files containing the data for each graph of data found in the manuscript including a separate .csv for corresponding statistics (files ending in _stats); 3) individual PDFs of graphs generated in GraphPad Prism; and 4) raw image files for microscopy and Western blots. These data demonstrate the principal findings for the manuscript that the protein SV2C: 1) enhances vesicular storage of dopamine and dopamine analogues (e.g., FFN206 and MPP+), and 2) confers neuroprotection against dopaminergic toxicity.
Description of the data and file structure
This dataset contains a GraphPad Prism file and four sub-folders labeled: 1) Datasheets, 2) Graphs, 3) Image files, and 4) Statistics. The GraphPad Prism file is inclusive of all of the data found in the manuscript and supplement with statistical analysis.
Within folder 1) Datasheets there are individual .csv files of the data used to generate the graphs found in the manuscript and on which statistics were performed.
In folder 2) Graphs there are PDFs of each individual graph generated from the data found in folder 1) Datasheets.
In folder 3) Image files there are the raw image files for microscopy and Western blot images found in the paper or used for quantitation.
In folder 4) Statistics there are individual .csv files corresponding to the data found in folder 1) Datasheets that contain the statistical analysis performed on the data. In the case of post-hoc tests (e.g., multiple comparisons), there is a second .csv file containing the results of the post-hoc tests.
Abbreviations
DA: dopamine
FFN: fluorescent false neurotransmitter
HEK: human embryonic kidney
ICC: immunocytochemistry
KO: knock-out
MPP+: 1-methyl-4-phenylpyridinium
SV2C: synaptic vesicle glycoprotein 2C
TH: tyrosine hydroxylase
VMAT2: vesicular monoamine transporter 2
WT: wild-type