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

Authors
  • Serdinšek, Tamara1
  • Lipovšek, Saša2, 3, 4, 5
  • Leitinger, Gerd5
  • But, Igor1
  • Stožer, Andraž2
  • Dolenšek, Jurij2, 3
  • 1 Clinic for Gynaecology and Perinatology, University Medical Centre Maribor, Ljubljanska 5, Maribor, 2000, Slovenia , Maribor (Slovenia)
  • 2 University of Maribor, Taborska ulica 8, Maribor, 2000, Slovenia , Maribor (Slovenia)
  • 3 University of Maribor, Koroška cesta 160, Maribor, 2000, Slovenia , Maribor (Slovenia)
  • 4 Faculty of Chemistry and Chemical Engineering, Smetanova ulica 17, University of Maribor, Maribor, 2000, Slovenia , Maribor (Slovenia)
  • 5 Medical University of Graz, Neue Stiftingtalstrasse 6, Graz, 8010, Austria , Graz (Austria)
Type
Published Article
Journal
Scientific Reports
Publisher
Springer Nature
Publication Date
Feb 14, 2020
Volume
10
Issue
1
Identifiers
DOI: 10.1038/s41598-020-59337-0
Source
Springer Nature
License
Green

Abstract

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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