Turner Heather N, Landry Christian, Galko Michael J
Department of Genetics, UT MD Anderson Cancer Center; Neuroscience Program, Graduate School of Biomedical Sciences at Houston; Section of Neurobiology, University of Southern California.
ProDev Engineering.
J Vis Exp. 2017 Apr 3(122):55568. doi: 10.3791/55568.
How organisms sense and respond to noxious temperatures is still poorly understood. Further, the mechanisms underlying sensitization of the sensory machinery, such as in patients experiencing peripheral neuropathy or injury-induced sensitization, are not well characterized. The genetically tractable Drosophila model has been used to study the cells and genes required for noxious heat detection, which has yielded multiple conserved genes of interest. Little is known however about the cells and receptors important for noxious cold sensing. Although, Drosophila does not survive prolonged exposure to cold temperatures (≤10 ºC), and will avoid cool, preferring warmer temperatures in behavioral preference assays, how they sense and possibly avoid noxious cold stimuli has only recently been investigated. Here we describe and characterize the first noxious cold (≤10 ºC) behavioral assay in Drosophila. Using this tool and assay, we show an investigator how to qualitatively and quantitatively assess cold nociceptive behaviors. This can be done under normal/healthy culture conditions, or presumably in the context of disease, injury or sensitization. Further, this assay can be applied to larvae selected for desired genotypes, which might impact thermosensation, pain, or nociceptive sensitization. Given that pain is a highly conserved process, using this assay to further study thermal nociception will likely glean important understanding of pain processes in other species, including vertebrates.
生物体如何感知并应对有害温度仍知之甚少。此外,感觉机制敏化的潜在机制,比如在患有周围神经病变的患者或损伤诱导敏化的患者中,尚未得到充分表征。具有遗传易处理性的果蝇模型已被用于研究有害热检测所需的细胞和基因,这产生了多个值得关注的保守基因。然而,对于有害冷感觉重要的细胞和受体却知之甚少。虽然果蝇在长时间暴露于低温(≤10ºC)下无法存活,并且在行为偏好试验中会避开凉爽环境,更喜欢较温暖的温度,但它们如何感知并可能避开有害冷刺激直到最近才被研究。在此,我们描述并表征了果蝇中首个有害冷(≤10ºC)行为试验。利用这个工具和试验,我们向研究人员展示了如何定性和定量评估冷伤害感受行为。这可以在正常/健康的培养条件下进行,或者推测也可以在疾病、损伤或敏化的背景下进行。此外,该试验可以应用于选择了所需基因型的幼虫,这些基因型可能会影响温度感觉、疼痛或伤害感受敏化。鉴于疼痛是一个高度保守的过程,使用这个试验进一步研究热伤害感受可能会让我们对包括脊椎动物在内的其他物种的疼痛过程有重要的了解。