McMackin Marissa Zubia, Lewin Matthew R, Tabuena Dennis R, Arreola F Eric, Moffatt Christopher, Fuse Megumi
Department of Biology, San Francisco State University, 1600 Holloway Ave, San Francisco, CA 94132, USA.
Institute for Biodiversity Science and Sustainability Director Center for Exploration and Travel Health, California Academy of Sciences, 55 Music Concourse Dr., Golden Gate Park, San Francisco, CA 94118, USA.
J Neurosci Methods. 2016 Jan 15;257:139-46. doi: 10.1016/j.jneumeth.2015.09.015. Epub 2015 Oct 9.
The hornworm Manduca sexta exhibits a defensive strike to noxious assaults, a response that is robust and is easily observed by experimenters. Von Frey filaments and methods typical for studying nociception in other animals were used to assess the strike response in M. sexta.
A series of von Frey filaments was applied to the body wall in ascending order and the data generated were used to determine the strike threshold by (i) the up-and-down method, (ii) the first response method, and (iii) the simplified up-and-down order method (SUDO). The effect of a noxious pinch on strike threshold was assessed.
To our knowledge none of these methods has been used on M. sexta previously, making the use of the up-and-down and SUDO methods the first in an invertebrate. The use of the first response method has been used in other invertebrates, and the method appears equally suited to M. sexta.
All three methods were successful in monitoring the threshold sensitivity to touch, which was lowered (sensitized) by tissue damage induced with a pinch. Sensitization lasted 19h.
All three methods of assessing nociception were successfully applied to quantify the defensive strike response in M. sexta, although the SUDO method required empirical assessment of which filament to start the test sequence with. The results revealed both short- and long-term sensitization. These methods should prove to be useful for quantifying sensitization in M. sexta.
烟草天蛾对有害攻击会表现出防御性攻击行为,这种反应强烈且易于实验者观察到。使用冯·弗里氏细丝及其他动物中研究伤害感受的典型方法来评估烟草天蛾的攻击反应。
将一系列冯·弗里氏细丝按升序施加于体壁,所产生的数据用于通过以下方法确定攻击阈值:(i)上下法,(ii)首次反应法,以及(iii)简化上下顺序法(SUDO)。评估有害挤压对攻击阈值的影响。
据我们所知,此前这些方法均未用于烟草天蛾,因此上下法和SUDO法的使用在无脊椎动物中尚属首次。首次反应法已在其他无脊椎动物中使用,且该方法似乎同样适用于烟草天蛾。
所有三种方法均成功监测到对触摸的阈值敏感性,有害挤压引起的组织损伤会使其降低(致敏)。致敏持续19小时。
所有三种评估伤害感受的方法均成功应用于量化烟草天蛾的防御性攻击反应,不过SUDO法需要凭经验评估开始测试序列时使用哪根细丝。结果显示了短期和长期致敏现象。这些方法应被证明对量化烟草天蛾的致敏有用。