Development of a murine pelvic ganglia-bladder preparation for the study of neural control of micturition

The urinary bladder collects and stores urine before releasing it at behaviorally appropriate times. Proper bladder function is controlled by autonomic nerve activity descending from the major pelvic ganglia (MPG). Current in vitro murine preparations used to study autonomic control of bladder function do not include the MPG and therefore limit our ability to study glia-neuron interactions within the MPG and their impact on bladder function. This study aimed to develop an MPG-bladder preparation in which MPG neurons can be optogenetically activated. Simultaneously, whole bladder contractions can be recorded as a readout for micturition function. To enable optogenetic activation of MPG parasympathetic neuronal activity, Chat-Cre::floxed-ChR2-EYFP mice were used in which the light-gated ion channel, namely channelrhodopsin 2 (ChR2) and enhanced yellow fluorescent protein (EYFP) were expressed in choline acetyltransferase (ChAT)-expressing parasympathetic neuronal bodies and efferent nerves. The MPG-bladder preparation was isolated from both male and female mice using different dissection protocols, and the intactness of the preparation were verified using whole tissue fluorescence imaging using spinning-disk confocal microscope following ethyl cinnamate (ECI)-based tissue clearing. To study the bladder contractile changes in response to optogenetic activation of parasympathetic efferent nerves, the preparation could be connected to a force transducer while optogenetic stimulation was applied to the MPG/bladder neck, leading to recordings of whole bladder contractility induced by parasympathetic efferent signal. Using this preparation, we observed frequency-dependent, parasympathetic nerve-induced whole bladder contractions. Overall, the development of this ex vivo mouse preparation will enable future hypothesis testing of the neuromodulatory roles of autonomic neurons and glia in bladder physiology, as well as the therapeutic potential of pharmacological agents for treating neurogenic bladder dysfunctions. This study was supported by CCTSI TM-T-24-263 to Xie & Calve. This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.