使用单一感器录音来检测臭虫的嗅觉神经元的反应，以信息化学物质
Summary
Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.
Abstract
昆虫嗅觉系统在环境中的检测信息化学物质中起重要作用。特别是，触角感器其中内部容纳单个或多个神经元，被认为是使在响应化学刺激的重大贡献。暴露于刺激后可直接记录动作电位的嗅觉感器，单感器记录（SSR）技术提供了强大的方法来研究昆虫化学刺激的神经反应。为臭虫，其是臭名昭著的人类寄生虫，多种类型的嗅觉感受器的已经表征。在这项研究中，我们证明了臭虫嗅觉感受器的神经反应两个化学刺激和剂量依赖反应使用SSR方法其中之一。这种方法使研究人员能够进行早期筛查有关臭虫的嗅觉感受器个人化学刺激，这将提供有价值的信息，为devel软件包opment新的臭虫引诱或驱虫剂和效益的臭虫的控制力度。
Introduction
常见的臭虫臭虫臭虫L（半翅目：Cimicidae），作为临时外寄生虫，是一种责任吸血昆虫，这意味着它们的生存，发育和繁殖需要从主机的血液来源，包括人类和动物1,2。虽然病毒的传播很少被报道，由于C.臭虫 ，由为患产生刺骨的滋扰，严重影响主机的生理和心理上3。引进和广泛使用化学杀虫剂，特别是滴滴涕，降低侵染的风险，由20世纪50年代的侵扰末端是在如此低的水平，他们不再是严重的公众关注。然而，许多可能的因素导致在床虱种群到回潮全世界，如杀虫剂的使用减少了，公众意识的下降，增加的行进活性，和耐药性的发展对杀虫剂4-9。</ P>
化学线索的环境中检测到，并通过嗅觉器官，如天线和上颌触须认可昆虫。对昆虫触角的嗅觉感受器在检测这些化学信号发挥了至关重要的作用。化学分子穿过角质层表面气孔进入触角角质层。气味结合蛋白在触角淋巴液绑定到这些化学分子和他们运到气味受体10。臭气物质受体和它们的共受体从神经膜的非选择性阳离子离子通道，这将在去极化一旦这些化学分子被气味受体11识别。
单感器记录（SSR）的开发，以检测所引起的化学或非化学刺激的应用的动作电位的胞外变化。通过将记录电极浸入感器淋巴液和参考电极入昆虫身体（通常是复眼或腹部）的其他部分，所述神经元的响应的燃烧率，以刺激可记录12。变化尖峰的数目代表该昆虫对特定刺激的敏感性。不同的身份和浓度的化学刺激将引起不同的神经反应，用不同的燃烧率和时间的结构，并且因此可以用于研究昆虫到特定化学品的编码处理。
对于常见的臭虫，有性形式分享嗅觉感受器的相同模式的触角：九槽的PEGÇ感器，29发丝般E（E1和E2）感器，而每个Dα，Dβ，Dγ顺利盯住一对感器13,14。作为多个神经元已经于每种类型感器的已经确定，这是不容易区分从安置在同一感器不同的神经元的动作电位，所以对于这个实验中托塔动作电位升数字被用于前和刺激后500毫秒周期期间计数的脱线。然后，从刺激前动作电位的数目中减去，并以量化在每秒 15尖峰每个单独感器中的变化的燃烧率乘以二的刺激后动作电位的数目。
Protocol
Representative Results
Discussion
的单感器记录技术已被广泛使用在测试昆虫的神经反应，如果苍蝇，蚊子和臭虫到环境中的不同的化学刺激。这些化学刺激往往溶解并稀释在一个共同的溶剂中，以制备不同剂量的治疗。然而，不同的溶剂可以产生完全不同的释放速度的刺激。在一些广泛的研究昆虫，例如果蝇 ， 冈比亚按蚊 ， 致倦库蚊和伊蚊已经过去的研究 通常使用石蜡油作为溶剂以溶解的?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
The project was supported by Award AAES 461Hatch/Multistate Grants ALA08-045 and ALA015-1-10026 to N.L.
Materials
| Tungsten wire | A-M SYSTEMS | #716500 | Used for preparing the electrode |
| KNO2 | Sigma | #310484 | Used for sharpening the tungsten wire |
| AC Power Supply | BK Precision | 1653A | Providing the voltage in sharpening the tungsten wire |
| Leica Z6 APO Microscope | Leica | 10447424 | Used for observing the sensilla on antennae |
| Simulus controller | Syntech | CS-55 | Used for controlling the stimulus application |
| 4-Channel USB Acquisition Controller | Syntech | IDAC-4 | Real-time on screen display of all signals before and during recording |
| Light Source | SCHOTT | A20500 | Providing light sources for observation |
| Micromanupulator | Leica | 115378 | Used for minor movement of electrode |
| Speaker | Juster | 95a | Connected with Acquisition Controller IDAC-4 and providing sound for the signal |
| Magnetic stand | Narishige | GJ-1 | Used to hold the reference electrode, stablized bed bug and stimulus delivery tube |
| TMC Vibration Isolation Table | TMC | 63-500 | Used for isolating the vibration from the equipments |
| Coverslip | Tedpella | 2225-1 | Used for holding the bed bug |
| Double-sided Tape | 3M | XT6110 | Used for stablizing the bed bug on the coverclip |
| Dental Wax | Dentakit | DK-R012 | Used for supporting the coverclip where bed bug is stablized |
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