Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods

The development of live-cell sensors for real-time measurement of signaling responses, with improved spatial and temporal resolution with respect to classical biochemical methods, has changed our understanding of cellular signaling. Examination of cAMP generation downstream activated GPCRs has shown...

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Detalles Bibliográficos
Publicado:	2019
Materias:	CRH receptors signaling CRHR1 Cyclic AMP Fluorescent flow cytometry FRET-based biosensors GPCR endocytosis Live-cell imaging
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0091679X_v149_n_p239_dosSantosClaro http://hdl.handle.net/20.500.12110/paper_0091679X_v149_n_p239_dosSantosClaro
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_0091679X_v149_n_p239_dosSantosClaro
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spelling	paper:paper_0091679X_v149_n_p239_dosSantosClaro2023-06-08T15:08:09Z Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods CRH receptors signaling CRHR1 Cyclic AMP Fluorescent flow cytometry FRET-based biosensors GPCR endocytosis Live-cell imaging The development of live-cell sensors for real-time measurement of signaling responses, with improved spatial and temporal resolution with respect to classical biochemical methods, has changed our understanding of cellular signaling. Examination of cAMP generation downstream activated GPCRs has shown that signaling responses can be short-lived (generated from the cell surface) or prolonged after receptor internalization. Class B secretin-like Corticotropin-releasing hormone receptor 1 (CRHR1) is a key player in stress pathophysiology. By monitoring real-time signaling in living cells, we uncovered cell context-dependent temporal characteristics of CRHR1-elicited cAMP responses and disclosed a specific link between cAMP generation and receptor signaling from internal compartments. We describe technical aspects and elaborate the protocols for cell line expression of Förster resonance energy transfer (FRET)-based biosensors to study the dynamics of cAMP and calcium signaling responses downstream activated CRHR1, live-cell imaging and analysis, and fluorescence flow cytometry to determine receptor levels at the cell surface. © 2019 Elsevier Inc. 2019 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0091679X_v149_n_p239_dosSantosClaro http://hdl.handle.net/20.500.12110/paper_0091679X_v149_n_p239_dosSantosClaro
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	CRH receptors signaling CRHR1 Cyclic AMP Fluorescent flow cytometry FRET-based biosensors GPCR endocytosis Live-cell imaging
spellingShingle	CRH receptors signaling CRHR1 Cyclic AMP Fluorescent flow cytometry FRET-based biosensors GPCR endocytosis Live-cell imaging Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
topic_facet	CRH receptors signaling CRHR1 Cyclic AMP Fluorescent flow cytometry FRET-based biosensors GPCR endocytosis Live-cell imaging
description	The development of live-cell sensors for real-time measurement of signaling responses, with improved spatial and temporal resolution with respect to classical biochemical methods, has changed our understanding of cellular signaling. Examination of cAMP generation downstream activated GPCRs has shown that signaling responses can be short-lived (generated from the cell surface) or prolonged after receptor internalization. Class B secretin-like Corticotropin-releasing hormone receptor 1 (CRHR1) is a key player in stress pathophysiology. By monitoring real-time signaling in living cells, we uncovered cell context-dependent temporal characteristics of CRHR1-elicited cAMP responses and disclosed a specific link between cAMP generation and receptor signaling from internal compartments. We describe technical aspects and elaborate the protocols for cell line expression of Förster resonance energy transfer (FRET)-based biosensors to study the dynamics of cAMP and calcium signaling responses downstream activated CRHR1, live-cell imaging and analysis, and fluorescence flow cytometry to determine receptor levels at the cell surface. © 2019 Elsevier Inc.
title	Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
title_short	Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
title_full	Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
title_fullStr	Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
title_full_unstemmed	Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
title_sort	assessing real-time signaling and agonist-induced crhr1 internalization by optical methods
publishDate	2019
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0091679X_v149_n_p239_dosSantosClaro http://hdl.handle.net/20.500.12110/paper_0091679X_v149_n_p239_dosSantosClaro
_version_	1768542980912709632

Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods

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