Author/Editor		Serdinšek, Tamara; Lipovšek Delakorda, Saška; Leitinger, Gerd; But, Igor; Stožer, Andraž; Dolenšek, Jurij
Title		A novel in situ approach to studying detrusor smooth muscle cells in mice
Type		članek
Vol. and No.		Letnik 10, št. art. no. 2685
Publication year		2020
Volume		str. 1-12 str.
ISSN		2045-2322 - Scientific reports
Language		eng
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%%M); causing activation of virtually all SMCs. Maximum calcium activity of SMCs was at about 25%%M, 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.
Keywords		bladder fluorescence imaging