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Mice In Mice

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Animals Female C57Bl/6 mice (Charles River, Wilmington, MA), born and housed in the Research Support Facility at the University of Kansas Medical Center, were used for all experiments. Mice received water and food ad libitum and were housed on a 12-hour light cycle (600 to 1800 hours). All research was approved by the University of Kansas Medical Center Institutional Animal Care and Use Committee (protocol numbers 2013-2150 and 2016-2344) in compliance with the National Institute of Health Guide for the Care and Use of Laboratory Animals. Experimental groups and ages Three separate cohorts of naïve and NMS mice were used to avoid confounding effects of additional stressors. In the first group, mice underwent urinary bladder distension …show more content…

Estrous cycle stage was estimated by determining the ratios of nucleated epithelial cells, cornified epithelial cells, and leukocytes. Estrous cycle estimations were made following a single observation and verification of estrus was made following at least 4 consecutive days of observation. No correlations between the estimated stage of estrous cycle and the results from behavioral assays or histological and molecular analysis were observed (data not shown). Urinary bladder distension Urinary bladder distension (UBD) was performed as previously described8. Briefly, under inhaled isoflurane (4% induction, 2% maintenance), wire electrodes (0.003” diameter; Grass Technologies, West Warwick, RI) were implanted into the abdominal musculature and the bladder was catheterized intravesically via the urethra. Anesthesia was lowered until hind limb reflexes, but not escape behaviors, were present (~0.9% isoflurane). Body temperature was maintained at approximately 37C. Gas flow from a compressed nitrogen tank was manually adjusted and monitored using a dual-stage low delivery pressure regulator (Matheson-Linweld, Kansas City, MO) and delivered via a custom-made distension control device (The University of Iowa Medical Instruments, Iowa City, IA). Following verification of stable responses to 60mmHg, each pressure (15,

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