scholarly journals Author response: Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain

2020 ◽  
Author(s):  
Lynne Chantranupong ◽  
Jessica L Saulnier ◽  
Wengang Wang ◽  
Drew R Jones ◽  
Michael E Pacold ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Author Response ◽  
Mammalian Brain ◽  
Metabolomic Profiling ◽  
Rapid Purification

scholarly journals Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain

2020 ◽  
Author(s):  
Lynne Chantranupong ◽  
Jessica L. Saulnier ◽  
Wengang Wang ◽  
Drew R. Jones ◽  
Michael E. Pacold ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Synaptic Vesicles ◽  
Brain Regions ◽  
Mammalian Brain ◽  
Metabolomic Profiling ◽  
Polar Metabolites ◽  
A Cell ◽  
Cell Type Specific ◽  
Rapid Purification ◽  
Activity Dependent

AbstractNeurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of synaptic vesicle content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at a high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. Finally, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission.

scholarly journals Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Lynne Chantranupong ◽  
Jessica L Saulnier ◽  
Wengang Wang ◽  
Drew R Jones ◽  
Michael E Pacold ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Synaptic Vesicles ◽  
Brain Regions ◽  
Mammalian Brain ◽  
Metabolomic Profiling ◽  
Polar Metabolites ◽  
A Cell ◽  
Cell Type Specific ◽  
Rapid Purification ◽  
Activity Dependent

Neurons communicate by the activity-dependent release of small-molecule neurotransmitters packaged into synaptic vesicles (SVs). Although many molecules have been identified as neurotransmitters, technical limitations have precluded a full metabolomic analysis of SV content. Here, we present a workflow to rapidly isolate SVs and to interrogate their metabolic contents at high-resolution using mass spectrometry. We validated the enrichment of glutamate in SVs of primary cortical neurons using targeted polar metabolomics. Unbiased and extensive global profiling of SVs isolated from these neurons revealed that the only detectable polar metabolites they contain are the established neurotransmitters glutamate and GABA. In addition, we adapted the approach to enable quick capture of SVs directly from brain tissue and determined the neurotransmitter profiles of diverse brain regions in a cell-type-specific manner. The speed, robustness, and precision of this method to interrogate SV contents will facilitate novel insights into the chemical basis of neurotransmission.

scholarly journals Author response: RIM-BP2 primes synaptic vesicles via recruitment of Munc13-1 at hippocampal mossy fiber synapses

2019 ◽  
Author(s):  
Marisa M Brockmann ◽  
Marta Maglione ◽  
Claudia G Willmes ◽  
Alexander Stumpf ◽  
Boris A Bouazza ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Mossy Fiber ◽  
Author Response

scholarly journals Author response: Frequency-dependent mobilization of heterogeneous pools of synaptic vesicles shapes presynaptic plasticity

2017 ◽  
Author(s):  
Frédéric Doussau ◽  
Hartmut Schmidt ◽  
Kevin Dorgans ◽  
Antoine M Valera ◽  
Bernard Poulain ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Author Response ◽  
Response Frequency ◽  
Frequency Dependent ◽  
Presynaptic Plasticity

scholarly journals Co-localization of different Neurotransmitter Transporters on the same Synaptic Vesicle is Bona-fide yet Sparse

2021 ◽  
Author(s):  
Neha Upmanyu ◽  
Jialin Jin ◽  
Marcelo Ganzella ◽  
Leon Boesche ◽  
Viveka Nand Malviya ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Glutamate Uptake ◽  
Quantitative Imaging ◽  
Small Population ◽  
Mammalian Brain ◽  
Content Type ◽  
Quantal Size ◽  
Vesicle Pools ◽  
Bona Fide ◽  
Vesicular Transporters

Vesicular transporters (VTs) define the type of neurotransmitter that synaptic vesicles (SVs) store and release. While certain neurons in mammalian brain release multiple transmitters, the prevalence, physiology of such pluralism and if the release occurs from same or distinct vesicle pools is not clear. Using quantitative imaging and biochemical approaches, we show that only a small population of neuronal SVs contain different VTs to accomplish corelease. Surprisingly, a highly diverse SV population (27 types) exist that express dual transporters suggesting corelease of diverse combinations of dual neurotransmitters, which includes the vesicle type that contains glutamate and zinc accounting for ∼34% of all SVs. Importantly, we demonstrate that transporter colocalization influences vesicular glutamate uptake leading to enhanced synaptic quantal size. Thus, localization of diverse transporters on single vesicles is bona-fide and the mechanism may underlie regulation of transmitter content, type and release in space and time.

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