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Studere pre-formet Fibril Indusert α-Synuclein akkumulering i primær embryonisk mus midbrain dopamin nevroner
Journal JoVE
Neurosciences
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Journal JoVE Neurosciences
Studying Pre-formed Fibril Induced α-Synuclein Accumulation in Primary Embryonic Mouse Midbrain Dopamine Neurons

Studere pre-formet Fibril Indusert α-Synuclein akkumulering i primær embryonisk mus midbrain dopamin nevroner

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

Chapitres

  • 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

Traduction automatique

Traduction automatique

Her presenterer vi en detaljert protokoll for å studere nevronal α-synuclein akkumulering i primære mus dopamin nevroner. Fosforylert α-synuclein aggregater i nevroner induseres med pre-formet α-synuclein fibriller. Automatisert avbildning av immunfluorescerende merkede celler og objektiv bildeanalyse gjør denne robuste protokollen egnet for middels til høy gjennomstrømningsscreening av legemidler som hemmer α-synuclein akkumulering.

Tags

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

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