ArticleFrontiers in molecular neuroscience2011
Concurrent imaging of synaptic vesicle recycling and calcium dynamics.
Article in Frontiers in molecular neuroscience, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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41 citing papers in PubMed, 72 citations in OpenAlex.
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- High-speed imaging of glutamate release with genetically encoded sensors.Nature protocols · 2019Article
- In vivo measurement of afferent activity with axon-specific calcium imaging.Nature neuroscience · 2018Article
- Newly produced synaptic vesicle proteins are preferentially used in synaptic transmission.The EMBO journal · 2018Article
- Identification of Fluorescent Small Molecule Compounds for Synaptic Labeling by Image-Based, High-Content Screening.ACS chemical neuroscience · 2018Article
- Proteolytic Processing of Neurexins by Presenilins Sustains Synaptic Vesicle Release.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2018Article
- Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis.Nature communications · 2017Article
- Sodium Channel β2 Subunits Prevent Action Potential Propagation Failures at Axonal Branch Points.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2017Article
- VGLUT2 Trafficking Is Differentially Regulated by Adaptor Proteins AP-1 and AP-3.Frontiers in cellular neuroscience · 2017Article
- Illuminating Brain Activities with Fluorescent Protein-Based Biosensors.Chemosensors (Basel, Switzerland) · 2017Article
- Genetically encoded indicators of neuronal activity.Nature neuroscience · 2016Review
- Visualizing Presynaptic Calcium Dynamics and Vesicle Fusion with a Single Genetically Encoded Reporter at Individual Synapses.Frontiers in synaptic neuroscience · 2016Article
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
Abstract
Synaptic transmission involves the calcium dependent release of neurotransmitter from synaptic vesicles. Genetically encoded optical probes emitting different wavelengths of fluorescent light in response to neuronal activity offer a powerful approach to understand the spatial and temporal relationship of calcium dynamics to the release of neurotransmitter in defined neuronal populations. To simultaneously image synaptic vesicle recycling and changes in cytosolic calcium, we developed a red-shifted reporter of vesicle recycling based on a vesicular glutamate transporter, VGLUT1-mOrange2 (VGLUT1-mOr2), and a presynaptically localized green calcium indicator, synaptophysin-GCaMP3 (SyGCaMP3) with a large dynamic range. The fluorescence of VGLUT1-mOr2 is quenched by the low pH of synaptic vesicles. Exocytosis upon electrical stimulation exposes the luminal mOr2 to the neutral extracellular pH and relieves fluorescence quenching. Reacidification of the vesicle upon endocytosis again reduces fluorescence intensity. Changes in fluorescence intensity thus monitor synaptic vesicle exo- and endocytosis, as demonstrated previously for the green VGLUT1-pHluorin. To monitor changes in calcium, we fused the synaptic vesicle protein synaptophysin to the recently improved calcium indicator GCaMP3. SyGCaMP3 is targeted to presynaptic varicosities, and exhibits changes in fluorescence in response to electrical stimulation consistent with changes in calcium concentration. Using real time imaging of both reporters expressed in the same synapses, we determine the time course of changes in VGLUT1 recycling in relation to changes in presynaptic calcium concentration. Inhibition of P/Q- and N-type calcium channels reduces calcium levels, as well as the rate of synaptic vesicle exocytosis and the fraction of vesicles released.
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