TIRF assays for real-time observation of microtubules and actin coassembly: Deciphering tau effects on microtubule/actin interplay - Archive ouverte HAL Accéder directement au contenu
Chapitre D'ouvrage Année : 2018

TIRF assays for real-time observation of microtubules and actin coassembly: Deciphering tau effects on microtubule/actin interplay

Résumé

Microtubule and actin cytoskeletons are key players in vital processes in cells. Although the importance of microtubule-actin interaction for cell development and function has been highlighted for years, the properties of these two cytoskeletons have been mostly studied separately. Thus we now need procedures to simultaneously assess actin and microtubule properties to decipher the basic mechanisms underlying microtubule-actin crosstalk. Here we describe an in vitro assay that allows the coassembly of both filaments and the real-time observation of their interaction by TIRF microscopy. We show how this assay can be used to demonstrate that tau, a neuronal microtubule-associated protein, is a bona fide actin-microtubule cross-linker. The procedure relies on the use of highly purified proteins and chemically passivated perfusion chambers. We present a step-by-step protocol to obtain actin and microtubule coassembly and discuss the major pitfalls. An ImageJ macro to quantify actin and microtubule interaction is also provided.
Fichier non déposé

Dates et versions

hal-02347026 , version 1 (05-11-2019)

Identifiants

Citer

Elea Prezel, Virginie Stoppin-Mellet, Auréliane Elie, Ninon Zala, Eric Denarier, et al.. TIRF assays for real-time observation of microtubules and actin coassembly: Deciphering tau effects on microtubule/actin interplay. Methods in au cell biology, 2018, ⟨10.1016/bs.mcb.2017.06.012⟩. ⟨hal-02347026⟩

Collections

UGA FRM
22 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More