Brenner Daniel S, Vogt Sherri K, Gereau Robert W
Washington University Pain Center and Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USA; Neuroscience Program, Washington University School of Medicine, St. Louis, MO 63110, USA; Medical Scientist Training Program, Washington University School of Medicine, St. Louis, MO 63110, USA.
Washington University Pain Center and Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
J Neurosci Methods. 2014 Oct 30;236:86-91. doi: 10.1016/j.jneumeth.2014.08.009. Epub 2014 Aug 13.
Adaptation to environmental temperature is essential for survival in seasonal areas. The mechanisms of adaptation have been studied in vitro, but it has not been quantified in vivo.
The extended Cold Plantar Assay (eCPA) cools the entire testing environment. Once the desired environmental temperature has been reached, a separate focal cold stimulus is applied to the hindpaw and the latency to withdrawal is recorded as a proxy for cold sensitivity.
Using this technique, we can test the cold responsiveness of freely behaving mice at ambient temperatures ranging from 5°C to 30°C. The responses are consistent and unambiguous, and the environmental temperatures generated are reproducible. We are also able to measure cold responsiveness as animals are in the process of adapting to cold environments.
COMPARISON WITH EXISTING METHOD(S): Existing methods, such as the dynamic cold plate and the 2-plate preference assay test how mice respond to cold environments, but cannot assess how the thresholds for response are changed by acclimation in cold environments. Additionally, the eCPA requires very little specialized equipment, can test many mice at the same time on one apparatus, and has an objective readout.
The extended Cold Plantar assay is a significant methodological improvement, allowing the assessment of cold responsiveness in freely behaving mice at a wide range of environmental temperature conditions and during cold adaptation.
适应环境温度对于季节性地区的生存至关重要。适应机制已在体外进行研究,但尚未在体内进行量化。
扩展冷足底试验(eCPA)可冷却整个测试环境。一旦达到所需的环境温度,就会对后爪施加单独的局部冷刺激,并记录撤回潜伏期作为冷敏感性的指标。
使用该技术,我们可以在5°C至30°C的环境温度下测试自由活动小鼠的冷反应性。反应一致且明确,所产生的环境温度具有可重复性。我们还能够在动物适应寒冷环境的过程中测量冷反应性。
现有方法,如动态冷板和双板偏好试验,测试小鼠对寒冷环境的反应,但无法评估冷环境适应如何改变反应阈值。此外,eCPA所需的专业设备很少,可以在一个仪器上同时测试许多小鼠,并且有客观的读数。
扩展冷足底试验是一项重大的方法学改进,能够在广泛的环境温度条件下以及在冷适应过程中评估自由活动小鼠的冷反应性。