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Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle

M(2) muscarinic acetylcholine receptors modulate cardiac rhythm via regulation of the inward potassium current. To increase our understanding of M(2) receptor physiology we used Total Internal Reflection Fluorescence Microscopy to visualize individual receptors at the plasma membrane of transformed...

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Autores principales: Nenasheva, Tatiana A., Neary, Marianne, Mashanov, Gregory I., Birdsall, Nigel J.M., Breckenridge, Ross A., Molloy, Justin E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3605596/
https://www.ncbi.nlm.nih.gov/pubmed/23357106
http://dx.doi.org/10.1016/j.yjmcc.2013.01.009
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author Nenasheva, Tatiana A.
Neary, Marianne
Mashanov, Gregory I.
Birdsall, Nigel J.M.
Breckenridge, Ross A.
Molloy, Justin E.
author_facet Nenasheva, Tatiana A.
Neary, Marianne
Mashanov, Gregory I.
Birdsall, Nigel J.M.
Breckenridge, Ross A.
Molloy, Justin E.
author_sort Nenasheva, Tatiana A.
collection PubMed
description M(2) muscarinic acetylcholine receptors modulate cardiac rhythm via regulation of the inward potassium current. To increase our understanding of M(2) receptor physiology we used Total Internal Reflection Fluorescence Microscopy to visualize individual receptors at the plasma membrane of transformed CHO(M2) cells, a cardiac cell line (HL-1), primary cardiomyocytes and tissue slices from pre- and post-natal mice. Receptor expression levels between individual cells in dissociated cardiomyocytes and heart slices were highly variable and only 10% of murine cardiomyocytes expressed muscarinic receptors. M(2) receptors were evenly distributed across individual cells and their density in freshly isolated embryonic cardiomyocytes was ~ 1 μm(− 2), increasing at birth (to ~ 3 μm(− 2)) and decreasing back to ~ 1 μm(− 2) after birth. M(2) receptors were primarily monomeric but formed reversible dimers. They diffused freely at the plasma membrane, moving approximately 4-times faster in heart slices than in cultured cardiomyocytes. Knowledge of receptor density and mobility has allowed receptor collision rate to be modeled by Monte Carlo simulations. Our estimated encounter rate of 5–10 collisions per second, may explain the latency between acetylcholine application and GIRK channel opening.
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spelling pubmed-36055962013-04-01 Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle Nenasheva, Tatiana A. Neary, Marianne Mashanov, Gregory I. Birdsall, Nigel J.M. Breckenridge, Ross A. Molloy, Justin E. J Mol Cell Cardiol Original Article M(2) muscarinic acetylcholine receptors modulate cardiac rhythm via regulation of the inward potassium current. To increase our understanding of M(2) receptor physiology we used Total Internal Reflection Fluorescence Microscopy to visualize individual receptors at the plasma membrane of transformed CHO(M2) cells, a cardiac cell line (HL-1), primary cardiomyocytes and tissue slices from pre- and post-natal mice. Receptor expression levels between individual cells in dissociated cardiomyocytes and heart slices were highly variable and only 10% of murine cardiomyocytes expressed muscarinic receptors. M(2) receptors were evenly distributed across individual cells and their density in freshly isolated embryonic cardiomyocytes was ~ 1 μm(− 2), increasing at birth (to ~ 3 μm(− 2)) and decreasing back to ~ 1 μm(− 2) after birth. M(2) receptors were primarily monomeric but formed reversible dimers. They diffused freely at the plasma membrane, moving approximately 4-times faster in heart slices than in cultured cardiomyocytes. Knowledge of receptor density and mobility has allowed receptor collision rate to be modeled by Monte Carlo simulations. Our estimated encounter rate of 5–10 collisions per second, may explain the latency between acetylcholine application and GIRK channel opening. Academic Press 2013-04 /pmc/articles/PMC3605596/ /pubmed/23357106 http://dx.doi.org/10.1016/j.yjmcc.2013.01.009 Text en © 2013 Elsevier Ltd. https://creativecommons.org/licenses/by/4.0/ Open Access under CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/) license
spellingShingle Original Article
Nenasheva, Tatiana A.
Neary, Marianne
Mashanov, Gregory I.
Birdsall, Nigel J.M.
Breckenridge, Ross A.
Molloy, Justin E.
Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title_full Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title_fullStr Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title_full_unstemmed Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title_short Abundance, distribution, mobility and oligomeric state of M(2) muscarinic acetylcholine receptors in live cardiac muscle
title_sort abundance, distribution, mobility and oligomeric state of m(2) muscarinic acetylcholine receptors in live cardiac muscle
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3605596/
https://www.ncbi.nlm.nih.gov/pubmed/23357106
http://dx.doi.org/10.1016/j.yjmcc.2013.01.009
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