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DOI: 10.3791/59546-v
Hyeonwi Son1, Ju Hwan Yang2, Hyun Joon Kim*1, Dong Kun Lee*3
1Department of Anatomy and Convergence Medical Sciences, Institute of Health Sciences, Bio Anti-aging Medical Research Center,Gyeongsang National University Medical School, 2Department of Biological Sciences,Pusan National University, 3Department of Physiology and Convergence Medical Sciences, Institute of Health Sciences,Gyeongsang National University Medical School
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本文提供了一个简化和标准化的协议,用于诱导长期固定小鼠的抑郁样行为使用约束剂。此外,还解释了验证抑郁症诱导的行为和生理技术。
首先,使用数字勒克斯仪表将房间设置为 200 lux 的照明。在测试前至少一周将鼠标放在单独的笼子里,并在实验前将鼠标放在测试室至少30分钟。轻轻握住鼠标尾部,避免使鼠标紧张,然后小心地将鼠标放在粗糙的表面上。
用一条小白毛巾盖住约束器,然后轻轻地将鼠标放在约束器的开口处,使鼠标自愿进入约束器。接下来,放置闭合点以尽可能紧绷鼠标,小心避免对身体造成伤害,如尾巴、脚和睾丸。最后,在接触约束剂期间,每48小时测量一次体重和食物摄入量。
在测试之前,让老鼠有两个饮水瓶,一个含有0.1摩尔蔗糖,另一个含有普通水48小时。24 小时后切换两个瓶子的位置,以减少侧偏产生的任何混淆。第三天,老鼠24小时缺水。
然后在实验当天,老鼠暴露在两个饮料瓶中6个小时。三小时后,切换水瓶的位置。记录蔗糖溶液和消耗水的体积,然后计算动物与蔗糖的亲和力。
最后,计算蔗糖偏好占蔗糖消费量占测试期间总液体消费量的百分比。首先将慢性约束压力诱导小鼠带进测试室至少30分钟,然后开始尾部悬浮测试。将房间光线设置为 50 lux 的昏暗条件。
接下来,将相机放在距离鼠标约 40 厘米的地方。使用胶带将鼠标牢固地从水平杆上悬住。鼠标放置在悬挂盒中间后开始录制,并连续观察行为变化六分钟。
如果鼠标试图爬上它的尾巴使用棍子或爬塞,以防止它这样做。实验结束时,将鼠标移到其主笼中,并小心地将胶带从尾巴上取下。最后,使用视频跟踪软件分析不动周期的累积时间。
结果显示,CIS组的体重下降,第四天食物摄入量明显减少。与对控相比,CIS组的蔗糖偏好下降,而固定时间和皮质酮显著增加。此外,谷氨酰胺补充饮食减轻抑郁,如行为,如体重,食物摄入量,蔗糖偏好,不动,和皮质酮。
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