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Pålidelig Identifikation af Living dopaminerge neuroner i midthjernen Cultures Brug RNA Sequencing og TH-promotor-drevet eGFP Expression
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
DOI:

10:54 min

February 10, 2017

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Chapters

  • 00:05Title
  • 00:59Isolation of Embryonic Midbrain
  • 02:55Dissociation and Plating of Cells
  • 05:34Harvesting Cultured Dopaminergic Neurons for RNA Sequencing
  • 07:04Generation, Purification and Amplification of First-strand cDNA
  • 09:13Results: RNA-seq Detection of Expressed Transcripts in Single Dopaminergic Neurons
  • 09:39Conclusion

Summary

Automatic Translation

Ved Parkinsons sygdom (PD), sorte substans (SNC) dopaminerge neuroner degenerere, hvilket fører til motorisk dysfunktion. Her rapporterer vi en protokol til dyrkning ventrale midthjernen neuroner fra en mus, der udtrykker eGFP drevet af en tyrosinhydroxylase (TH) promotorsekvens, høst individuelle fluorescerende neuroner fra kulturerne, og måling af deres transkriptom anvendelse af RNA-seq.

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