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Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis
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JoVE Journal Neuroscience
Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

DOI:
10.3791/62813-v

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08:59 min

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July 16, 2021

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, , ,
1Jefferson Weinberg ALS Center,Thomas Jefferson University

Chapters

  • 00:04Introduction
  • 00:52Microscope and Perfusion System Setup
  • 02:33Styryl Dye Imaging of Synaptic Vesicle Release
  • 04:30Fluorescence Imaging of Gcamp6m Calcium Transients
  • 05:36Image Analysis
  • 06:23Results: Real-Time Monitoring of the Dynamics of Presynaptic Calcium Influx and Synaptic Vesicle Membrane Fusion in In Vitro Cultured Neurons
  • 08:15Conclusion

Summary

Automatic Translation

Automatic Translation

Two related methods are described to visualize subcellular events required for synaptic transmission. These protocols enable the real-time monitoring of the dynamics of presynaptic calcium influx and synaptic vesicle membrane fusion using live-cell imaging of in vitro cultured neurons.

Tags

Keywords: Real-timeFluorescentSynapticAmyotrophic Lateral SclerosisSynaptic FunctionElectrophysiologyPrimary Rodent Neuronal CulturesInduced Pluripotent Stem CellsGCaMPStyryl DyeACSFPotassium ChlorideConfocal Imaging

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