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Related Experiment Video

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Microtubule Plus-End Dynamics Visualization in Huntington's Disease Model based on Human Primary Skin Fibroblasts

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History-dependent catastrophes regulate axonal microtubule behavior.

Tatiana Stepanova¹, Ihor Smal, Jeffrey van Haren

¹Department of Cell Biology and Genetics, Erasmus MC, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands.

Current Biology : CB

|May 18, 2010

Summary

Axonal microtubules grow at a constant speed and undergo programmed catastrophes. Microtubule-associated proteins CLIPs and EB1 regulate neuronal microtubule dynamics and turnover.

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Area of Science:

Cell Biology
Neuroscience
Biophysics

Background:

Microtubules in Chinese hamster ovary cells originate from the microtubule organizing center (MTOC) and grow towards the cell edge.
Axonal microtubules grow parallel to the plasma membrane without an MTOC, requiring different regulatory mechanisms.
Microtubule plus end tracking proteins (+TIPs) are crucial for regulating microtubule dynamics in neurons.

Purpose of the Study:

To investigate the regulation of microtubule dynamics in neuronal axons.
To determine the roles of specific +TIPs, CLIP-115, CLIP-170, and EB1, in axonal microtubule growth and stability.
To understand the mechanisms underlying microtubule catastrophes in axons.

Main Methods:

Utilized GFP-tagged +TIPs to visualize and track growing neuronal microtubules.
Employed fluorescence-based segmentation and tracking tools for quantitative analysis.
Performed protein depletion experiments to assess the function of CLIP-115, CLIP-170, and EB1.

Main Results:

Axonal microtubules exhibit constant average growth velocity and undergo catastrophes at random yet programmed positions.
CLIP-115 and CLIP-170 influence microtubule growth rate and distance in neurons, but not growth duration.
EB1 regulates microtubule growth rate, growth distance, and duration in neuroblastoma cells, aligning with in vitro findings.

Conclusions:

CLIPs modulate the axonal microtubule/tubulin ratio.
EB1 promotes microtubule growth and structural transitions at microtubule ends.
These +TIPs collectively regulate microtubule catastrophes and the turnover of +TIP binding sites in axons.