DOI: 10.5061/dryad.zpc866tdc
title: Synaptic vesicle glycoprotein 2C enhances vesicular storage of dopamine and counters dopaminergic toxicity

description: 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.
description: # 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