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Undersøgelse Præ-dannet Fibril Induceret α-Synuclein ophobning i primær embryonale mus Midbrain dopamin neuroner
JoVE Journal
Neurociência
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JoVE Journal Neurociência
Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons

Undersøgelse Præ-dannet Fibril Induceret α-Synuclein ophobning i primær embryonale mus Midbrain dopamin neuroner

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

Capítulos

  • 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

Tadução automática

Tadução automática

Her præsenterer vi en detaljeret protokol til undersøgelse af neuronal α-synucleinakkumulering i primære muse dopaminneuroner. Fosforylerede α-synucleinaggregater i neuroner induceres med præformede α-synuclein fibrils. Automatiseret billeddannelse af immunfluorescent mærket celler og upartisk billedanalyse gør denne robuste protokol egnet til medium-til-høj gennemløb screening af lægemidler, der hæmmer α-synuclein akkumulering.

Tags

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

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