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A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice

The aim of our study was to develop a novel approach to investigating mouse detrusor smooth muscle cell (SMC) physiological activity, utilizing an acute tissue dissection technique and confocal calcium imaging. The bladder of a sacrificed adult female NMRI mouse was dissected. We used light and tran...

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Autores principales: Serdinšek, Tamara, Lipovšek, Saša, Leitinger, Gerd, But, Igor, Stožer, Andraž, Dolenšek, Jurij
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7021722/
https://www.ncbi.nlm.nih.gov/pubmed/32060298
http://dx.doi.org/10.1038/s41598-020-59337-0
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author Serdinšek, Tamara
Lipovšek, Saša
Leitinger, Gerd
But, Igor
Stožer, Andraž
Dolenšek, Jurij
author_facet Serdinšek, Tamara
Lipovšek, Saša
Leitinger, Gerd
But, Igor
Stožer, Andraž
Dolenšek, Jurij
author_sort Serdinšek, Tamara
collection PubMed
description The aim of our study was to develop a novel approach to investigating mouse detrusor smooth muscle cell (SMC) physiological activity, utilizing an acute tissue dissection technique and confocal calcium imaging. The bladder of a sacrificed adult female NMRI mouse was dissected. We used light and transmission electron microscopy to assess morphology of SMCs within the tissue. Calcium imaging in individual SMCs was performed using confocal microscopy during stimulation with increasing concentrations of carbamylcholine (CCh). SMCs were identified according to their morphology and calcium activity. We determined several parameters describing the SMC responses: delays to response, recruitment, relative activity, and contraction of the tissue. CCh stimulation revealed three different SMC phenotypes: spontaneously active SMCs with and without CCh-enhanced activity and SMCs with CCh-induced activity only. SMCs were recruited into an active state in response to CCh-stimulation within a narrow range (1–25 µM); causing activation of virtually all SMCs. Maximum calcium activity of SMCs was at about 25 µM, which coincided with a visible tissue contraction. Finally, we observed shorter time lags before response onsets with higher CCh concentrations. In conclusion, our novel in situ approach proved to be a robust and reproducible method to study detrusor SMC morphology and physiology.
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spelling pubmed-70217222020-02-24 A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice Serdinšek, Tamara Lipovšek, Saša Leitinger, Gerd But, Igor Stožer, Andraž Dolenšek, Jurij Sci Rep Article The aim of our study was to develop a novel approach to investigating mouse detrusor smooth muscle cell (SMC) physiological activity, utilizing an acute tissue dissection technique and confocal calcium imaging. The bladder of a sacrificed adult female NMRI mouse was dissected. We used light and transmission electron microscopy to assess morphology of SMCs within the tissue. Calcium imaging in individual SMCs was performed using confocal microscopy during stimulation with increasing concentrations of carbamylcholine (CCh). SMCs were identified according to their morphology and calcium activity. We determined several parameters describing the SMC responses: delays to response, recruitment, relative activity, and contraction of the tissue. CCh stimulation revealed three different SMC phenotypes: spontaneously active SMCs with and without CCh-enhanced activity and SMCs with CCh-induced activity only. SMCs were recruited into an active state in response to CCh-stimulation within a narrow range (1–25 µM); causing activation of virtually all SMCs. Maximum calcium activity of SMCs was at about 25 µM, which coincided with a visible tissue contraction. Finally, we observed shorter time lags before response onsets with higher CCh concentrations. In conclusion, our novel in situ approach proved to be a robust and reproducible method to study detrusor SMC morphology and physiology. Nature Publishing Group UK 2020-02-14 /pmc/articles/PMC7021722/ /pubmed/32060298 http://dx.doi.org/10.1038/s41598-020-59337-0 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Serdinšek, Tamara
Lipovšek, Saša
Leitinger, Gerd
But, Igor
Stožer, Andraž
Dolenšek, Jurij
A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title_full A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title_fullStr A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title_full_unstemmed A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title_short A Novel in situ Approach to Studying Detrusor Smooth Muscle Cells in Mice
title_sort novel in situ approach to studying detrusor smooth muscle cells in mice
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7021722/
https://www.ncbi.nlm.nih.gov/pubmed/32060298
http://dx.doi.org/10.1038/s41598-020-59337-0
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