Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments06:53

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

5.5K
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...
5.5K
Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy (3D-SIM)07:40

Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy (3D-SIM)

2.4K
The present protocol describes a method to visualize and measure actin rings and other components of the membrane periodic skeleton of the axon initial segment using cultured rat hippocampal neurons and 3D-structured illumination microscopy...
2.4K
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends12:20

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

14.9K
Microtubules are inherently unstable polymers, and their switching between growth and shortening is stochastic and difficult to control. Here we describe protocols using segmented microtubules with photoablatable stabilizing caps. Depolymerization of segmented microtubules can be triggered with high temporal and spatial resolution, thereby assisting analysis of motions with the disassembling microtubule...
14.9K
In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons05:58

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

9.9K
This protocol discusses the live dissection of Drosophila larvae for the purpose of imaging the movement of GFP tagged axonal vesicles on microtubule...
9.9K
Feedback Inhibition00:46

Feedback Inhibition

56.9K
Biochemical reactions are occurring constantly in cells, converting starting substances to different products, usually with the help of enzymes that speed the reactions. Without enzymes, it would take far too long for most reactions to occur to be useful to the cell!
56.9K
ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly16:33

ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly

13.0K
A method to obtain nanofibers and complex nanostructures from single or multiple extracellular matrix proteins is described. This method uses protein-surface interactions to create free-standing protein-based materials with tunable composition and architecture for use in a variety of tissue engineering and biotechnology...
13.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

SPACA9 and MNMIP1 bridge the seam of spermatid manchette microtubules.

The EMBO journal·2026
Same author

Slow but steady: Molecular mechanisms controlling microtubule growth in centrioles and cilia.

The Journal of cell biology·2026
Same author

Author Correction: Development of the fluorescent probe CenSpark for labeling centrioles and cilia.

Nature chemical biology·2026
Same author

Microtubule organization and molecular architecture of ciliary basal bodies in multiciliated airway cells.

Current biology : CB·2026
Same author

Development of the fluorescent probe CenSpark for labeling centrioles and cilia.

Nature chemical biology·2026
Same author

Cooperative multivalency converts disorder into rods, resolving a paradox in cellular architecture.

bioRxiv : the preprint server for biology·2026
Same journal

Fast-conducting mechanonociceptors uniquely engage reflexive and affective pain circuitry to drive protective responses.

Neuron·2026
Same journal

Sparse component analysis: A method that uncovers separable computations within neural population activity.

Neuron·2026
Same journal

Spatiomolecular mapping reveals anatomical organization of heterogeneous cell types in the human nucleus accumbens.

Neuron·2026
Same journal

TGF-β1-induced endothelial transcytosis drives blood-brain barrier leakage during aging.

Neuron·2026
Same journal

Image space opens up for visual neuroscience.

Neuron·2026
Same journal

Septal GLP-1 receptors control alcohol taking and seeking.

Neuron·2026
See all related articles

Related Experiment Video

Updated: Jan 20, 2026

Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments
06:53

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

Published on: February 12, 2021

5.5K

Feedback-Driven Assembly of the Axon Initial Segment.

Amélie Fréal1, Dipti Rai2, Roderick P Tas2

  • 1Cell Biology, Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, the Netherlands; Department of Axonal Signaling, Netherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and Sciences, Meibergdreef 47, 1105 BA Amsterdam, the Netherlands.

Neuron
|September 3, 2019
PubMed
Summary
This summary is machine-generated.

Ankyrin-G scaffolds proteins to recruit microtubules, enabling Neurofascin-186 accumulation at the axon initial segment (AIS) for neuronal polarity. This reveals feedback mechanisms essential for AIS assembly.

Keywords:
axon initial segmentaxonal transportendocytosismicrotubules

More Related Videos

Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy 3D-SIM
07:40

Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy 3D-SIM

Published on: February 11, 2022

2.4K
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
12:20

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

Published on: March 15, 2014

14.9K

Related Experiment Videos

Last Updated: Jan 20, 2026

Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments
06:53

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

Published on: February 12, 2021

5.5K
Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy 3D-SIM
07:40

Measuring Properties of the Membrane Periodic Skeleton of the Axon Initial Segment using 3D-Structured Illumination Microscopy 3D-SIM

Published on: February 11, 2022

2.4K
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
12:20

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

Published on: March 15, 2014

14.9K

Area of Science:

  • Neuroscience
  • Cell Biology
  • Molecular Biology

Background:

  • The axon initial segment (AIS) is critical for neuronal function, requiring complex protein assembly.
  • Key proteins like Ankyrin-G and TRIM46 are involved in AIS formation, but their interactions are unclear.

Purpose of the Study:

  • To elucidate the cooperative mechanisms of Ankyrin-G, TRIM46, and Neurofascin-186 in AIS assembly.
  • To understand how these proteins establish neuronal polarity and action potential generation.

Main Methods:

  • In vitro reconstitution assays.
  • Cellular assays to observe protein interactions and localization.
  • Microtubule dynamics and transport studies.

Main Results:

  • Ankyrin-G recruits TRIM46-decorated microtubules to the plasma membrane via interactions with EB proteins and Neurofascin-186.
  • TRIM46 stabilizes microtubules and promotes retrograde transport of Neurofascin-186 to the proximal axon.
  • Ankyrin-G prevents Neurofascin-186 endocytosis, leading to its accumulation and reinforcing the Ankyrin-G scaffold.

Conclusions:

  • A feedback loop involving Ankyrin-G, TRIM46, and Neurofascin-186 drives AIS assembly and stability.
  • This mechanism is crucial for establishing neuronal polarity and function.
  • Understanding these interactions provides insights into neurodevelopmental and neurological disorders.