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Analysis of Microtubules and Microtubule-Organizing Center at the Immune Synapse

  • Noelia Blas-Rus
  • Eugenio Bustos-Morán
  • Francisco Sánchez-MadridEmail author
  • Noa B. Martín-Cófreces
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1584)

Abstract

The immune synapse (IS) is a specialized structure that enables cell-cell communication between immune cells. As such, it involves direct cell-to-cell contact. It is sustained by cytoskeletal components that allow the intracellular polarization of different organelles and the surface re-organization of signaling and adhesion receptors. The tubulin-based cytoskeleton is a key player in IS formation and signaling. We describe methods to analyze through Western blot and microscopy analysis the polarization to the IS of the centrosome, also known as microtubule-organizing center (MTOC), the dynamics of microtubule growth and polymerization from the MTOC to the IS and the activation of signaling molecules.

Key words

Immune synapse Cytoskeleton Signaling T Cell receptor Mitochondria Centrosome Microtubules 

Notes

Acknowledgments

We thank Dr. Miguel Vicente Manzanares for critical reading of the manuscript. Optical microscopy experimentation has been conducted at the Microscopy & Dynamic Imaging Unit of the CNIC (Centro Nacional de Investigaciones Cardiovasculares) and at the Microscopy Facility of the IIS-IP (Instituto Investigación Sanitaria-Instituto Princesa), Madrid, Spain. This study was supported by grants SAF2014-55579-R from the Spanish Ministry of Economy and Competitiveness, INDISNET-S2011/BMD-2332 from the Comunidad de Madrid, ERC-2011-AdG 294340-GENTRIS Red Cardiovascular and RD 12-0042-0056 from Instituto Salud Carlos III (ISCIII). The Centro Nacional de Investigaciones Cardiovasculares (CNIC, Spain) is supported by the Spanish Ministry of Science and Innovation and the Pro-CNIC Foundation.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Noelia Blas-Rus
    • 1
  • Eugenio Bustos-Morán
    • 2
  • Francisco Sánchez-Madrid
    • 1
    • 2
    Email author
  • Noa B. Martín-Cófreces
    • 1
    • 2
  1. 1.Servicio de Inmunología, Hospital Universitario de la PrincesaUniversidad Autónoma de Madrid, Instituto Investigación Sanitaria Princesa (IIS-IP)MadridSpain
  2. 2.Vascular PathophysiologyAreaCentro Nacional Investigaciones Cardiovasculares (CNIC)MadridSpain

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