CNO-hM3D mediated perturbation of noradrenergic DBH-Cre neurons whole animal plethysmography recordings

Plethysmography on conscious, unrestrained mice was carried out as described on 6-12 week old adult animals with at least 12 animals in both the experimental and control group for a given experiment. Mice were subjected to a five-day habituation protocol with each day consisting of several minutes of handling, temperature taken by rectal probe, intraperitoneal saline injection, and 30 minutes in the plethysmography chamber. Mice were then tested within one week of the last day of conditioning. On the day of testing, mice were taken from their home cage, weighed, and rectal temperature was taken. Animals were then placed into an airtight, temperature controlled (~32°C) plethysmography chamber and allowed to acclimate for at least 20 minutes in room air (21% O2/79% N2) conditions. After acclimation and measurement under room air, the chamber gas was switched to a hypercapnic mixture of 5% CO2/21% O2/74% N2 for 20 minutes. Chamber gas was then switched back to room air for 20 minutes. The mice were briefly removed for rectal temperature measurement and intra-peritoneal injection of clozapine-N-oxide (CNO, National Institute of Mental Health Chemical Synthesis and Drug Supply Program) dissolved in saline (0.1 mg/mL) for an effective concentration 1 mg/kg. The animal was returned to the chamber for another 20 minutes of room air, 20 minutes of hypercapnia, and 20 minutes of room air. The animal was then removed from the chamber and rectal temperature was taken immediately afterwards and again 30 minutes after the termination of the experiment. The animal was placed in its own cage during these 30 minutes at the ambient room temperature (~23°C). Individual recordings for all animals expressing hM3D receptors in NA neurons are in this repository.

本研究对6~12周龄的成年小鼠开展清醒无束缚状态下的体积描记法（Plethysmography）检测，每项实验的实验组与对照组均至少包含12只动物。实验前小鼠需接受为期5天的适应方案：每日进行数分钟的抓取抚触、直肠探针测温、腹腔注射生理盐水，并在体积描记舱内停留30分钟。于最后一次适应训练结束后的1周内开展测试。测试当日，将小鼠从饲养笼中取出，称量体重并测量直肠体温。随后将动物放入密闭温控（约32℃）的体积描记舱，在常压空气（21%O₂/79%N₂）环境中适应至少20分钟。完成常压空气下的适应与测量后，将舱内气体切换为5%CO₂/21%O₂/74%N₂的高碳酸混合气体，持续20分钟；随后再次切换回常压空气，维持20分钟。将小鼠短暂取出，测量直肠体温，并腹腔注射溶于生理盐水（0.1mg/mL）的氯氮平-N-氧化物（clozapine-N-oxide, CNO，美国国立精神卫生研究所化学合成与药物供应项目），给药有效浓度为1mg/kg。将动物放回舱内，依次进行20分钟常压空气暴露、20分钟高碳酸血症暴露及20分钟常压空气暴露。随后将动物移出舱外，立即测量直肠体温，并于实验结束30分钟后再次测量体温；此期间将动物置于单独饲养笼中，环境温度为室温（约23℃）。本数据集仓库收录了所有在去甲肾上腺素能（NA）神经元中表达hM3D受体的动物的个体检测记录。