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Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons

Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons

DOI:
10.3791/61118-v

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10:03 min

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August 16, 2020

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, , , ,
1Institute of Biotechnology, HiLIFE,University of Helsinki, 2Neuroscience Center, HiLIFE,University of Helsinki, 3Department of Brain Biochemistry, Maj Institute of Pharmacology,Polish Academy of Sciences

Chapters

  • 00:04Introduction
  • 01:11Primary Embryonic Midbrain Culture Plate Preparation
  • 01:53Primary E13.5 Embryonic Midbrain Culture Setup
  • 04:32Seeding with Pre-Formed Fibrils (PFFs) and Semi-automated Immunofluorescence Imaging
  • 06:21High-Content Image Analysis
  • 07:42Results: Representative Midbrain Culture Characterization and Analysis
  • 09:13Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a detailed protocol to study neuronal α-synuclein accumulation in primary mouse dopamine neurons. Phosphorylated α-synuclein aggregates in neurons are induced with pre-formed α-synuclein fibrils. Automated imaging of immunofluorescently labeled cells and unbiased image analysis make this robust protocol suitable for medium-to-high throughput screening of drugs that inhibit α-synuclein accumulation.

Tags

Keywords: Alpha-synucleinPreformed FibrilsPrimary Dopamine NeuronsParkinson’s DiseaseNeurodegenerative DiseaseProtein AccumulationCell CultureHigh-throughput ScreeningImage AnalysisNeuron FunctionMolecular Mechanisms

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