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Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation
Journal JoVE
Neurosciences
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Journal JoVE Neurosciences
Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation

Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation

DOI:
10.3791/57884-v

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09:16 min

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June 26, 2018

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, ,
1Bio@SNS Laboratory,Scuola Normale Superiore, 2International School for Advanced Studies (SISSA/ISAS)

Chapitres

  • 00:04Titre
  • 00:36Isolation of Microsomes-associated Alpha-synuclein Aggregates from Diseased A53T Alpha-synuclein Transgenic Mic
  • 02:30Western Blot
  • 04:32Neurons Treatment
  • 05:40Immunofluorescence
  • 07:45Results: Time-dependent Induction of Alpha-synuclein Deposition After Administration of Microsomes-associated Alpha-synuclein Aggregates
  • 08:35Conclusion

Summary

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The goal of this protocol is to provide a cell-based system that replicates the formation of alpha-synuclein aggregates in vivo. Intracellular alpha-synuclein inclusions are seeded in primary neurons by the internalization and propagation of exogenous administered native microsomes-associated alpha-synuclein aggregates isolated from diseased alpha-synuclein transgenic mice.

Tags

Keywords: Alpha-synucleinAggregationFibrilsPrimary NeuronsExogenous AdministrationMicrosomesWestern BlotUltracentrifugationHomogenization BufferCell Model

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