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Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments
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JoVE Journal Neuroscience
Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments

Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments

DOI:
10.3791/61411-v

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06:53 min

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February 12, 2021

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,
1School of Basic Medical Sciences,Xi’an Jiaotong University, 2Department of Cell Biology,Duke University, 3Department of Biochemistry,Duke University

Chapters

  • 00:04Introduction
  • 00:24Culture Hippocampal Neurons
  • 02:37Disruption of AIS by Knockout of Ankyrin-G at an Early Stage of Neuron Development
  • 04:17Quantification of Axon Initial Segment
  • 04:58Results: A Full Rescue of AIS by 480kDa AnkG in ANK3-E22/23-flox Neurons Transfected with Cre

Summary

Automatic Translation

Automatic Translation

Here, we described a protocol to quantitatively study the assembly and structure of the axon initial segments (AIS) of hippocampal neurons that lack pre-assembled AIS due to the absence of a giant ankyrin-G.

Tags

Primary Cultured Hippocampal NeuronsAxon Initial SegmentsAnkyrin-GCRE BFP DNAAnkyrin-G GFP DNATransfectionDissectionTrypsin DigestionTriturationNeuronal CultureGlial Cell Feeder Dishes

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