Artículo

dos Santos Claro, P.A.; Inda, C.; Armando, N.G.; Piazza, V.G.; Attorresi, A.; Silberstein, S.; Shukla A.K."Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods" (2019) Methods in Cell Biology. 149:239-257
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Abstract:

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.

Registro:

Documento: Artículo
Título:Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods
Autor:dos Santos Claro, P.A.; Inda, C.; Armando, N.G.; Piazza, V.G.; Attorresi, A.; Silberstein, S.; Shukla A.K.
Filiación:Instituto de Investigación en Biomedicina de Buenos Aires (IBioBA)-CONICET-Partner Institute of the Max Planck Society, Buenos Aires, Argentina
Departamento Fisiología, Biología Molecular y Celular, Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Buenos Aires, Argentina
Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Buenos Aires, Argentina
Departamento de Química Biológica, Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina
Palabras clave:CRH receptors signaling; CRHR1; Cyclic AMP; Fluorescent flow cytometry; FRET-based biosensors; GPCR endocytosis; Live-cell imaging
Año:2019
Volumen:149
Página de inicio:239
Página de fin:257
DOI: http://dx.doi.org/10.1016/bs.mcb.2018.08.009
Handle:http://hdl.handle.net/20.500.12110/paper_0091679X_v149_n_p239_dosSantosClaro
Título revista:Methods in Cell Biology
Título revista abreviado:Methods Cell Biol.
ISSN:0091679X
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0091679X_v149_n_p239_dosSantosClaro

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Citas:

---------- APA ----------
dos Santos Claro, P.A., Inda, C., Armando, N.G., Piazza, V.G., Attorresi, A., Silberstein, S. & Shukla A.K. (2019) . Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods. Methods in Cell Biology, 149, 239-257.
http://dx.doi.org/10.1016/bs.mcb.2018.08.009
---------- CHICAGO ----------
dos Santos Claro, P.A., Inda, C., Armando, N.G., Piazza, V.G., Attorresi, A., Silberstein, S., et al. "Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods" . Methods in Cell Biology 149 (2019) : 239-257.
http://dx.doi.org/10.1016/bs.mcb.2018.08.009
---------- MLA ----------
dos Santos Claro, P.A., Inda, C., Armando, N.G., Piazza, V.G., Attorresi, A., Silberstein, S., et al. "Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods" . Methods in Cell Biology, vol. 149, 2019, pp. 239-257.
http://dx.doi.org/10.1016/bs.mcb.2018.08.009
---------- VANCOUVER ----------
dos Santos Claro, P.A., Inda, C., Armando, N.G., Piazza, V.G., Attorresi, A., Silberstein, S., et al. Assessing real-time signaling and agonist-induced CRHR1 internalization by optical methods. Methods Cell Biol. 2019;149:239-257.
http://dx.doi.org/10.1016/bs.mcb.2018.08.009