Feedback-Driven Assembly of the Axon Initial Segment

Amélie Fréal; Dipti Rai; Roderick P Tas; Xingxiu Pan; Eugene A Katrukha; Dieudonnée van de Willige; Riccardo Stucchi; Amol Aher; Chao Yang; A F Maarten Altelaar; Karin Vocking; Jan Andries Post; Martin Harterink; Lukas C Kapitein; Anna Akhmanova; Casper C Hoogenraad

doi:10.1016/j.neuron.2019.07.029

Feedback-Driven Assembly of the Axon Initial Segment

Amélie Fréal, Dipti Rai, Roderick P Tas, Xingxiu Pan, Eugene A Katrukha, Dieudonnée van de Willige, Riccardo Stucchi, Amol Aher, Chao Yang, A F Maarten Altelaar, Karin Vocking, Jan Andries Post, Martin Harterink, Lukas C Kapitein, Anna Akhmanova, Casper C Hoogenraad

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The axon initial segment (AIS) is a unique neuronal compartment that plays a crucial role in the generation of action potential and neuronal polarity. The assembly of the AIS requires membrane, scaffolding, and cytoskeletal proteins, including Ankyrin-G and TRIM46. How these components cooperate in AIS formation is currently poorly understood. Here, we show that Ankyrin-G acts as a scaffold interacting with End-Binding (EB) proteins and membrane proteins such as Neurofascin-186 to recruit TRIM46-positive microtubules to the plasma membrane. Using in vitro reconstitution and cellular assays, we demonstrate that TRIM46 forms parallel microtubule bundles and stabilizes them by acting as a rescue factor. TRIM46-labeled microtubules drive retrograde transport of Neurofascin-186 to the proximal axon, where Ankyrin-G prevents its endocytosis, resulting in stable accumulation of Neurofascin-186 at the AIS. Neurofascin-186 enrichment in turn reinforces membrane anchoring of Ankyrin-G and subsequent recruitment of TRIM46-decorated microtubules. Our study reveals feedback-based mechanisms driving AIS assembly.

Original language	English
Pages (from-to)	P305-321.E8
Journal	Neuron
Volume	104
Issue number	2
DOIs	https://doi.org/10.1016/j.neuron.2019.07.029
Publication status	Published - 23 Oct 2019

Keywords

axon initial segment
axonal transport
microtubules
endocytosis

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1-s2.0-S0896627319306555-mainFinal published version, 8.54 MBLicence: CC BY

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title = "Feedback-Driven Assembly of the Axon Initial Segment",

abstract = "The axon initial segment (AIS) is a unique neuronal compartment that plays a crucial role in the generation of action potential and neuronal polarity. The assembly of the AIS requires membrane, scaffolding, and cytoskeletal proteins, including Ankyrin-G and TRIM46. How these components cooperate in AIS formation is currently poorly understood. Here, we show that Ankyrin-G acts as a scaffold interacting with End-Binding (EB) proteins and membrane proteins such as Neurofascin-186 to recruit TRIM46-positive microtubules to the plasma membrane. Using in vitro reconstitution and cellular assays, we demonstrate that TRIM46 forms parallel microtubule bundles and stabilizes them by acting as a rescue factor. TRIM46-labeled microtubules drive retrograde transport of Neurofascin-186 to the proximal axon, where Ankyrin-G prevents its endocytosis, resulting in stable accumulation of Neurofascin-186 at the AIS. Neurofascin-186 enrichment in turn reinforces membrane anchoring of Ankyrin-G and subsequent recruitment of TRIM46-decorated microtubules. Our study reveals feedback-based mechanisms driving AIS assembly.",

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author = "Am{\'e}lie Fr{\'e}al and Dipti Rai and Tas, {Roderick P} and Xingxiu Pan and Katrukha, {Eugene A} and {van de Willige}, Dieudonn{\'e}e and Riccardo Stucchi and Amol Aher and Chao Yang and Altelaar, {A F Maarten} and Karin Vocking and Post, {Jan Andries} and Martin Harterink and Kapitein, {Lukas C} and Anna Akhmanova and Hoogenraad, {Casper C}",

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T1 - Feedback-Driven Assembly of the Axon Initial Segment

AU - Fréal, Amélie

AU - Rai, Dipti

AU - Tas, Roderick P

AU - Pan, Xingxiu

AU - Katrukha, Eugene A

AU - van de Willige, Dieudonnée

AU - Stucchi, Riccardo

AU - Aher, Amol

AU - Yang, Chao

AU - Altelaar, A F Maarten

AU - Vocking, Karin

AU - Post, Jan Andries

AU - Harterink, Martin

AU - Kapitein, Lukas C

AU - Akhmanova, Anna

AU - Hoogenraad, Casper C

PY - 2019/10/23

Y1 - 2019/10/23

N2 - The axon initial segment (AIS) is a unique neuronal compartment that plays a crucial role in the generation of action potential and neuronal polarity. The assembly of the AIS requires membrane, scaffolding, and cytoskeletal proteins, including Ankyrin-G and TRIM46. How these components cooperate in AIS formation is currently poorly understood. Here, we show that Ankyrin-G acts as a scaffold interacting with End-Binding (EB) proteins and membrane proteins such as Neurofascin-186 to recruit TRIM46-positive microtubules to the plasma membrane. Using in vitro reconstitution and cellular assays, we demonstrate that TRIM46 forms parallel microtubule bundles and stabilizes them by acting as a rescue factor. TRIM46-labeled microtubules drive retrograde transport of Neurofascin-186 to the proximal axon, where Ankyrin-G prevents its endocytosis, resulting in stable accumulation of Neurofascin-186 at the AIS. Neurofascin-186 enrichment in turn reinforces membrane anchoring of Ankyrin-G and subsequent recruitment of TRIM46-decorated microtubules. Our study reveals feedback-based mechanisms driving AIS assembly.

AB - The axon initial segment (AIS) is a unique neuronal compartment that plays a crucial role in the generation of action potential and neuronal polarity. The assembly of the AIS requires membrane, scaffolding, and cytoskeletal proteins, including Ankyrin-G and TRIM46. How these components cooperate in AIS formation is currently poorly understood. Here, we show that Ankyrin-G acts as a scaffold interacting with End-Binding (EB) proteins and membrane proteins such as Neurofascin-186 to recruit TRIM46-positive microtubules to the plasma membrane. Using in vitro reconstitution and cellular assays, we demonstrate that TRIM46 forms parallel microtubule bundles and stabilizes them by acting as a rescue factor. TRIM46-labeled microtubules drive retrograde transport of Neurofascin-186 to the proximal axon, where Ankyrin-G prevents its endocytosis, resulting in stable accumulation of Neurofascin-186 at the AIS. Neurofascin-186 enrichment in turn reinforces membrane anchoring of Ankyrin-G and subsequent recruitment of TRIM46-decorated microtubules. Our study reveals feedback-based mechanisms driving AIS assembly.

KW - axon initial segment

KW - axonal transport

KW - microtubules

KW - endocytosis

U2 - 10.1016/j.neuron.2019.07.029

DO - 10.1016/j.neuron.2019.07.029

M3 - Article

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SN - 0896-6273

VL - 104

SP - P305-321.E8

JO - Neuron

JF - Neuron

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ER -

Feedback-Driven Assembly of the Axon Initial Segment

Abstract

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