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Deciphering cellular signals in adult mouse sinoatrial node cells

Sinoatrial node (SAN) cells are the pacemakers of the heart. This study describes a method for culturing and infection of adult mouse SAN cells with FRET-based biosensors that can be exploited to examine signaling events. SAN cells cultured in media with blebbistatin or (S)-nitro-blebbistatin retain...

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Autores principales: Reddy, Gopireddy R., Ren, Lu, Thai, Phung N., Caldwell, Jessica L., Zaccolo, Manuela, Bossuyt, Julie, Ripplinger, Crystal M., Xiang, Yang K., Nieves-Cintrón, Madeline, Chiamvimonvat, Nipavan, Navedo, Manuel F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749457/
https://www.ncbi.nlm.nih.gov/pubmed/35036877
http://dx.doi.org/10.1016/j.isci.2021.103693
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author Reddy, Gopireddy R.
Ren, Lu
Thai, Phung N.
Caldwell, Jessica L.
Zaccolo, Manuela
Bossuyt, Julie
Ripplinger, Crystal M.
Xiang, Yang K.
Nieves-Cintrón, Madeline
Chiamvimonvat, Nipavan
Navedo, Manuel F.
author_facet Reddy, Gopireddy R.
Ren, Lu
Thai, Phung N.
Caldwell, Jessica L.
Zaccolo, Manuela
Bossuyt, Julie
Ripplinger, Crystal M.
Xiang, Yang K.
Nieves-Cintrón, Madeline
Chiamvimonvat, Nipavan
Navedo, Manuel F.
author_sort Reddy, Gopireddy R.
collection PubMed
description Sinoatrial node (SAN) cells are the pacemakers of the heart. This study describes a method for culturing and infection of adult mouse SAN cells with FRET-based biosensors that can be exploited to examine signaling events. SAN cells cultured in media with blebbistatin or (S)-nitro-blebbistatin retain their morphology, protein distribution, action potential (AP) waveform, and cAMP dynamics for at least 40 h. SAN cells expressing targeted cAMP sensors show distinct β-adrenergic-mediated cAMP pools. Cyclic GMP, protein kinase A, Ca(2+)/CaM kinase II, and protein kinase D in SAN cells also show unique dynamics to different stimuli. Heart failure SAN cells show a decrease in cAMP and cGMP levels. In summary, a reliable method for maintaining adult mouse SAN cells in culture is presented, which facilitates studies of signaling networks and regulatory mechanisms during physiological and pathological conditions.
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spelling pubmed-87494572022-01-14 Deciphering cellular signals in adult mouse sinoatrial node cells Reddy, Gopireddy R. Ren, Lu Thai, Phung N. Caldwell, Jessica L. Zaccolo, Manuela Bossuyt, Julie Ripplinger, Crystal M. Xiang, Yang K. Nieves-Cintrón, Madeline Chiamvimonvat, Nipavan Navedo, Manuel F. iScience Article Sinoatrial node (SAN) cells are the pacemakers of the heart. This study describes a method for culturing and infection of adult mouse SAN cells with FRET-based biosensors that can be exploited to examine signaling events. SAN cells cultured in media with blebbistatin or (S)-nitro-blebbistatin retain their morphology, protein distribution, action potential (AP) waveform, and cAMP dynamics for at least 40 h. SAN cells expressing targeted cAMP sensors show distinct β-adrenergic-mediated cAMP pools. Cyclic GMP, protein kinase A, Ca(2+)/CaM kinase II, and protein kinase D in SAN cells also show unique dynamics to different stimuli. Heart failure SAN cells show a decrease in cAMP and cGMP levels. In summary, a reliable method for maintaining adult mouse SAN cells in culture is presented, which facilitates studies of signaling networks and regulatory mechanisms during physiological and pathological conditions. Elsevier 2021-12-25 /pmc/articles/PMC8749457/ /pubmed/35036877 http://dx.doi.org/10.1016/j.isci.2021.103693 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Reddy, Gopireddy R.
Ren, Lu
Thai, Phung N.
Caldwell, Jessica L.
Zaccolo, Manuela
Bossuyt, Julie
Ripplinger, Crystal M.
Xiang, Yang K.
Nieves-Cintrón, Madeline
Chiamvimonvat, Nipavan
Navedo, Manuel F.
Deciphering cellular signals in adult mouse sinoatrial node cells
title Deciphering cellular signals in adult mouse sinoatrial node cells
title_full Deciphering cellular signals in adult mouse sinoatrial node cells
title_fullStr Deciphering cellular signals in adult mouse sinoatrial node cells
title_full_unstemmed Deciphering cellular signals in adult mouse sinoatrial node cells
title_short Deciphering cellular signals in adult mouse sinoatrial node cells
title_sort deciphering cellular signals in adult mouse sinoatrial node cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749457/
https://www.ncbi.nlm.nih.gov/pubmed/35036877
http://dx.doi.org/10.1016/j.isci.2021.103693
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