title: GPCRs and actin-cytoskeleton dynamics.
abstract: A multitude of physiological processes regulated by G protein-coupled receptors (GPCRs) signaling are accomplished by the participation of active rearrangements of the cytoskeleton. In general, it is common that a cross talk occurs among networks of microfilaments, microtubules, and intermediate filaments in order to reach specific cell responses. In particular, actin-cytoskeleton dynamics regulate processes such as cell shape, cell division, cell motility, and cell polarization, among others. This chapter describes the current knowledge about the regulation of actin-cytoskeleton dynamic by diverse GPCR signaling pathways, and also includes some protocols combining immunofluorescence and confocal microscopy for the visualization of the different rearrangements of the actin-cytoskeleton. We report how both the S1P-GPCR/G12/13/Rho/ROCK and glucagon-GPCR/Gs/cAMP axes induce differential actin-cytoskeleton rearrangements in epithelial cells. We also show that specific actin-binding molecules, like phalloidin and LifeAct, are very useful to analyze F-actin reorganization by confocal microscopy, and also that both molecules show similar results in fixed cells, whereas the anti-actin antibody is useful to detect both the G- and F-actin, as well as their compartmentalization. Thus, it is highly recommended to utilize different approaches to investigate the regulation of actin dynamics by GPCR signaling, with the aim to get a better picture of the phenomenon under study.
authors.affiliation: Departamento de Biología Celular y Desarrollo, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México D.F., Mexico.
authors.forename: Genaro
authors.initials: G
authors.lastname: Vázquez-Victorio
authors.affiliation: Departamento de Farmacobiología, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico.
authors.forename: Claudia
authors.initials: C
authors.lastname: González-Espinosa
authors.affiliation: Departamento de Farmacobiología, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico.
authors.forename: Zyanya P
authors.initials: ZP
authors.lastname: Espinosa-Riquer
authors.affiliation: Departamento de Biología Celular y Desarrollo, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México D.F., Mexico.
authors.forename: Marina
authors.initials: M
authors.lastname: Macías-Silva
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002460:Cell Line; D007382:Intermediate Filaments / Q000502:physiology / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D055503:Protein Multimerization; D051381:Rats; D020239:Receptor Cross-Talk; D043562:Receptors, G-Protein-Coupled / Q000502:physiology*; D015398:Signal Transduction
chemical_list: D043562:Receptors, G-Protein-Coupled
keywords: Actin; Cytoskeleton; G protein; G(12/13); GEFs; GPCR; Lifeact; Microfilaments; Phalloidin; Rho