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Quantitative Monitoring of GPCR-Mediated Spatiotemporal IP3 Dynamics Using Confocal Fluorescence Microscopy.
Xu, Xuehua; Ha, HyunGee; Brzostowski, Joseph; Jin, Tian.
Afiliación
  • Xu X; Chemotaxis Signaling Section, Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Rockville, MD, USA. xxu@niaid.nih.gov.
  • Ha H; Chemotaxis Signaling Section, Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Rockville, MD, USA.
  • Brzostowski J; Imaging Core Facility, Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Rockville, MD, USA.
  • Jin T; Chemotaxis Signaling Section, Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Rockville, MD, USA.
Methods Mol Biol ; 2814: 195-207, 2024.
Article en En | MEDLINE | ID: mdl-38954207
ABSTRACT
Activation of G protein-coupled receptors upon chemoattractant stimulation induces activation of multiple signaling pathways. To fully understand how these signaling pathway coordinates to achieve directional migration of neutrophils, it is essential to determine the dynamics of the spatiotemporal activation profile of signaling components at the level of single living cells. Here, we describe a detailed methodology for monitoring and quantitatively analyzing the spatiotemporal dynamics of 1,4,5-inositol trisphosphate (IP3) in neutrophil-like HL60 cells in response to various chemoattractant fields by applying Förster resonance energy transfer (FRET) fluorescence microscopy.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Inositol 1,4,5-Trifosfato / Microscopía Confocal / Transferencia Resonante de Energía de Fluorescencia / Receptores Acoplados a Proteínas G / Microscopía Fluorescente Límite: Humans Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Inositol 1,4,5-Trifosfato / Microscopía Confocal / Transferencia Resonante de Energía de Fluorescencia / Receptores Acoplados a Proteínas G / Microscopía Fluorescente Límite: Humans Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos