The project was supported by Award AAES 461Hatch/Multistate Grants ALA08-045 and ALA015-1-10026 to N.L.

1.准备仪器，刺激方案和臭虫的<ol><li>制备在20ml的瓶子50％KNO 2溶液（重量/体积）。 </li><li>通过反复浸渍钨电极和流出溶液的锐化2钨微电极KNO 2溶液在5 V。 <ol><li>大致通过浸渍钨丝和移出KNO 2溶液的约10毫米，在2骤降/秒的速度约5分钟，从而可以大大消耗钨丝的前端锐化钨丝。 </li><li>微妙锐化电极浸渍在进出溶液的金属丝端头为约1毫米，在2骤降/秒的速度进行至少1分钟，以便使了微细且鲜明点电极。检查电极尖端直径在显微镜下频频，直至达到0.2-0.5微米，这应该是好到足以击穿的臭虫嗅觉感受器的角质层。 注意:当手动锐厄宁所述电极，所述钨丝送入KNO 2溶液的浸渍速度不是恒定的所有时间。随着越来越多的实践中，这是很容易保持一个相对恒定的速度在锐化电极。磨刀时间也是不确定取决于电极应该怎么罚金。这里，电极尖端〜0.2微米的直径足以刺穿嗅觉感受器。 </li></ol></li><li>每个从整齐化合物二甲亚砜（DMSO）中的化学刺激稀释至1:10体积/体积的初始浓度作为原液。创建一系列十进​​制稀释液根据多少剂量在实验用DMSO需要，再次，从每个原液每个化学。这里，使用10％的（+） - β蒎烯和桉叶油。 </li><li>将不喂食物，七天后饲养的成年臭虫（男性或女性）从英尺。在T使用迪克斯殖民地（从海恩斯在肯塔基大学博士赠送）他尝试在培养皿。 注意:对于放置在培养皿臭虫没有确切的数字。它可以是几个或很多。 </li></ol> 2.臭虫触角准备<ol><li>麻醉冰（2-3分钟）的臭虫。 </li><li>解决这两个天线和虫体在显微镜盖玻片用双面胶带，并用细剪刀去除腿部。 </li><li>用一个小针轻轻触碰天线，以便把它们粘在磁带上稳步推进。 </li><li>休息盖玻片针对牙科蜡的一个小球（〜1厘米直径）以方便操纵，并将其调整到一个适当的角度（〜90°）为记录电极（ 图1）。 </li><li>一旦固定，将臭虫立体显微镜下，打开冷光源，调整光照强度，直到天线清晰呈现，并聚焦显微镜上的臭虫天线在高放大倍率的第二鞭毛（720X） 。 注意:在实验中使用照明的强度是不定量，这真的取决于实验者的眼睛的感觉照度。 </li></ol> 3.单感器记录<ol><li>连接前置放大器（10X）与信号采集控制器，其与所述计算机进行信号记录和可视化连接。打开电脑，并启动该软件， 例如 ，AutoSpike32并单击"录制"模式从菜单栏中。然后选择"波"，以开始录制波信号。 注:平线从左侧重复运行到显示器的，现在应该是可见的。这里，记录窗口持续40秒。最大波记录为10秒。选择的采样速率是96000和数字采样率是240有0％偏移和无滤波，没有整流为记录信号。在该软件的所有这些参数的设置可以根据需要进行修改。 </li> &lt;利&gt;打开连接到前置放大器，其用于呈现调色模式从触角感器的神经元的反应的扬声器。 </li><li>将参比电极进入稳定臭虫的腹部。 注意:参考电极举行通过金属站在磁力附着到空气表。 </li><li>后参考电极被连接到臭虫的腹部，将记录电极，其连接到前置放大器和操纵由显微，朝向臭虫的天线的后端。 </li><li>当记录电极与天线的右顶端接触，开关在显微镜并定位在低倍率的电极。 </li><li>调整记录电极，同时逐渐增大放大倍数，直到两个电极和触角感器是在同一平面上，并在显微镜下清晰可见。 注意:这个时候，显微镜通常LY最高放大倍率。 </li><li>将记录电极到使用显微感器的轴，去得更深一些，如果比动作电位的背景噪声高。 </li><li>一旦清楚动作电位是从所记录的感器观察，填充微量吸有10％（+） - β蒎烯。使用微量沉积由10％（+），10微升等分试样β - 蒎烯到滤纸条（〜3×15毫米）放置在玻璃巴斯德吸管内。 <ol><li>加载的吸移管连接至刺激控制器的脉冲流管的出口与吸移管的尖端放入小孔面向天线管。 </li></ol></li><li>当所有这些连接已经稳定，抑制了刺激控制的脚踏开关来实现刺激（0.5升/分）的0.5秒粉扑进入连​​续加湿空气流。动作电位的记录将被同时启动时footswitch是郁闷。录制过程将持续10秒开始刺激前1秒。 </li><li>计数动作电位离线两个500毫秒期间，一前一刺激后。减去500毫秒后的刺激来自于前500毫秒记录的自发活动期间在穗率的任何变化，并通过它们乘以2转换成计数尖峰/ s的常规规模。 </li></ol> 4.更换刺激的SSR <ol><li>在吸移管β蒎烯到臭虫触角并记录该特定加臭剂的响应10秒钟，然后将吸管除去 - 一旦脚踏开关被触发，提供10％的（+）。 </li><li>标号另一个新的吸移管与0.001％桉叶油素进行测试。放置一小片滤纸到其上10微升的刺激得到了应用，到新的吸管。 </li><li>等待2-5分钟，直到刺激是完全vapori捷思在玻璃吸管。连接吸管到脉冲流管的出口。 </li><li>将移液管尖端到面向天线管的小孔。踩下脚踏开关，启动10秒的录音。 </li><li>断开吸管，并准备另一吸管0.01％桉。 </li><li>测试桉叶油素的剂量（0.001％至10％）的触角感受器的所有其余观察的剂量依赖性反应。从最测试稀释到最低稀释剂量。 </li></ol>

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The authors have nothing to disclose.

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?...

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。&lt;/ P&gt; 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。

<table border="0" cellpadding="0" cellspacing="0" width="1064">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:299px;">Tungsten wire</td>
			<td style="width:139px;">A-M SYSTEMS</td>
			<td style="width:117px;">#716500</td>
			<td style="width:511px;">Used for preparing the electrode</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">KNO2</td>
			<td>Sigma</td>
			<td>#310484</td>
			<td>Used for sharpening the tungsten wire</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">AC Power Supply</td>
			<td>BK Precision</td>
			<td>1653A</td>
			<td>Providing the voltage in sharpening the tungsten wire</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Leica Z6 APO Microscope</td>
			<td>Leica</td>
			<td>10447424</td>
			<td>Used for observing the sensilla on antennae</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Simulus controller</td>
			<td>Syntech</td>
			<td>CS-55</td>
			<td>Used for controlling the stimulus application</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4-Channel USB Acquisition Controller</td>
			<td>Syntech</td>
			<td>IDAC-4</td>
			<td>Real-time on screen display of all signals before and during recording</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Light Source</td>
			<td>SCHOTT</td>
			<td>A20500</td>
			<td>Providing light sources for observation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Micromanupulator</td>
			<td>Leica</td>
			<td>115378</td>
			<td>Used for minor movement of electrode</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Speaker</td>
			<td>Juster</td>
			<td>95a</td>
			<td>Connected with Acquisition Controller IDAC-4 and providing sound for the signal</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Magnetic stand</td>
			<td>Narishige</td>
			<td>GJ-1</td>
			<td>Used to hold the reference electrode, stablized bed bug and stimulus delivery tube</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">TMC Vibration Isolation Table</td>
			<td>TMC</td>
			<td>63-500</td>
			<td>Used for isolating the vibration from the equipments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Coverslip</td>
			<td>Tedpella</td>
			<td>2225-1</td>
			<td>Used for holding the bed bug</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Double-sided Tape</td>
			<td>3M</td>
			<td>XT6110</td>
			<td>Used for stablizing the bed bug on the coverclip</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dental Wax</td>
			<td>Dentakit</td>
			<td>DK-R012</td>
			<td>Used for supporting the coverclip where bed bug is stablized&#160;</td>
		</tr>
	</tbody>
</table>

using single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals

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。

单感器录音昆虫化学生态学和神经生理学的研究使用了强大的侦查手段。调查昆虫不同的挥发性化合物，特别是那些想到的神经反应是生态环境关系到昆虫的生存和发展，不仅为我们提供了宝贵的见解昆虫嗅觉过程中，同时也开启了充满希望的新途径，它可能会导致到有用的新试剂的发展，防治病虫害。 常见的臭虫，作为臭?...

Watch this Scientific Journal Video about Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals at JoVE.com

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

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.

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

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

using-single-sensillum-recording-to-detect-olfactory-neuron-responses

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

9.8K 次观看.  Auburn University. 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.

视频: Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

在昆虫单感器记录果蝇按蚊</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

科研

神经科学

Analyzing Responses of Mouse Olfactory Sensory Neurons Using the Air-phase Electroolfactogram Recording

Animals depend on olfaction for many critical behaviors, such as finding food sources, avoiding predators, and identifying conspecifics for mating and other social interactions. The electroolfactogram (EOG) recording is an informative, easy to conduct, and reliable method to assay olfactory function at the level of the olfactory epithelium. Since the 1956 description of the EOG by Ottoson in frogs1, the EOG recording has been applied in many vertebrates including salamanders, rabbits, rats, mice, and humans (reviewed by Scott and Scott-Johnson, 2002, ref. 2). The recent advances in genetic modification in mice have rekindled interest in recording the EOG for physiological characterization of olfactory function in knock-out and knock-in mice. EOG recordings have been successfully applied to demonstrate the central role of olfactory signal transduction components3-8, and more recently to characterize the contribution of certain regulatory mechanisms to OSN responses9-12.
Odorant detection occurs at the surface of the olfactory epithelium on the cilia of OSNs, where a signal transduction cascade leads to opening of ion channels, generating a current that flows into the cilia and depolarizes the membrane13. The EOG is the negative potential recorded extracellularly at the surface of the olfactory epithelium upon odorant stimulation, resulting from a summation of the potential changes caused by individual responsive OSNs in the recording field2. Comparison of the amplitude and kinetics of the EOG thus provide valuable information about how genetic modification and other experimental manipulations influence the molecular signaling underlying the OSN response to odor. 
Here we describe an air-phase EOG recording on a preparation of mouse olfactory turbinates. Briefly, after sacrificing the mouse, the olfactory turbinates are exposed by bisecting the head along the midline and removing the septum. The turbinate preparation is then placed in the recording setup, and a recording electrode is placed at the surface of the olfactory epithelium on one of the medial turbinates. A reference electrode is electrically connected to the tissue through a buffer solution. A continuous stream of humidified air is blown over the surface of the epithelium to keep it moist. The vapor of odorant solutions is puffed into the stream of humidified air to stimulate the epithelium. Responses are recorded and digitized for further analysis.

The electroolfactogram (EOG) recording is an informative, easy-to-conduct, and reliable way of assessing olfactory function at the level of the olfactory epithelium. This protocol describes a recording setup, mouse tissue preparation, data collection, and basic data analysis.

鼠标采用的空气相Electroolfactogram录制的嗅觉感官神经元响应分析

Animals depend on olfaction for many critical behaviors, such as finding food sources, avoiding predators, and identifying conspecifics for mating and ...

Recording Behavioral Responses to Reflection in Crayfish

Social behavior depends on sensory input from the visual, mechanical and olfactory systems. One important issue concerns the relative roles of each sensory modality in guiding behavior. The role of visual inputs has been examined by isolating visual stimuli from mechanical and chemosensory stimuli. In some studies (Bruski &amp; Dunham, 1987: Delgado-Morales et al., 2004) visual inputs have been removed with blindfolds or low light intensity, and effects of remaining sensory modalities have been elucidated. An alternative approach is to study the effects of visual inputs in the absence of any appropriate mechanical and chemosensory cues. This approach aims to identify the exclusive role of visual inputs. 
We have used two methods to provide visual stimuli to crayfish without providing chemical and mechanical cues. In one method, crayfish are videotaped in an aquarium where half of the walls are covered in mirrors to provide a reflective environment, and the other half are covered in a non-reflective (matte finish) plastic. This gives the crayfish a choice between reflective and non-reflective environments. The reflective environment provides visual cues in the form of reflected images of the crayfish as it moves throughout half of the tank; these visual cues are missing from the non-reflective half of the tank. An alternative method is to videotape the behavior of crayfish in an aquarium separated by a smaller chamber at each end, with a crayfish in one small chamber providing visual cues and an inert object in the opposite small chamber providing visual input from a non-moving, non-crayfish source. 
Our published results indicate that responses of crayfish to the reflective environment depend on socialization and dominance rank. Socialized crayfish spent more time in the reflective environment and exhibited certain behaviors more frequently there than in the non-reflective environment; isolated crayfish showed no such differences. Crayfish that were housed in same-sex pairs developed a social rank of either dominant or subordinate. Responses to reflection differed between dominant and subordinate crayfish (May &amp; Mercier, 2006; May &amp; Mercier, 2007). Dominant crayfish spent more time on the reflective side, entered reflective corners more frequently and spent more time in reflective corners compared to the non-reflective side. Subordinate crayfish walked in reverse more often on the reflective side than on the non-reflective side. Preliminary data suggest similar effects from visual cues provided by a crayfish in a small adjoining chamber (May et al., 2008).

We have developed two methods for studying effects of visual cues on behavior in the absence of tactile and chemical cues. One method involves videotaping responses of crayfish to reflective walls in an aquarium; the other examines effects of visual inputs provided by a live crayfish behind a transparent partition.

小龙虾的反思记录行为反应

Social behavior depends on sensory input from the visual, mechanical and olfactory systems.  One important issue concerns the relative roles of each ...

Using a Comparative Species Approach to Investigate the Neurobiology of Paternal Responses

A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors. Using animal models to accomplish this goal, many methodological strategies require invasive techniques to manipulate the intensity of the behavior of interest (e.g., lesion methods, pharmacological manipulations, microdialysis techniques, genetically-engineered animal models). The utilization of a comparative species approach allows researchers to take advantage of naturally occurring differences in response strategies existing in closely related species. In our lab, we use two species of the Peromyscus genus that differ in paternal responses. The male California deer mouse (Peromyscus californicus) exhibits the same parental responses as the female whereas its cousin, the common deer mouse (Peromyscus maniculatus) exhibits virtually no nurturing/parental responses in the presence of pups. Of specific interest in this article is an exploration of the neurobiological factors associated with the affiliative social responses exhibited by the paternal California deer mouse. Because the behavioral neuroscience approach is multifaceted, the following key components of the study will be briefly addressed: the identification of appropriate species for this type of research; data collection for behavioral analysis; preparation and sectioning of the brains; basic steps involved in immunocytochemistry for the quantification of vasopressin-immunoreactivity; the use of neuroimaging software to quantify the brain tissue; the use of a microsequencing video analysis to score behavior and, finally, the appropriate statistical analyses to provide the most informed interpretations of the research findings.

The comparative species approach allows behavioral neuroscientists to explore various neurobiological factors associated with specific behaviors viewed as characteristic of a specific animal model. Taking advantage of naturally occurring differences in behavior between closely related species, this technique doesn&#x2019;t require invasive techniques to manipulate the expression of the behavior.

使用方法比较物种调查父亲的反应神经生物学

A goal of behavioral neuroscience is to identify underlying neurobiological factors that regulate specific behaviors.  Using animal models to accomplish ...

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex behaviors such as food choice, predator, conspecific and mate recognition and other socially relevant cues. Olfactory receptor neurons (ORNs) are located in the dorsal part of the nasal cavity embedded in the olfactory epithelium. These bipolar neurons send an axon to the olfactory bulb (see Fig. 1, Reisert &amp; Zhao1, originally published in the Journal of General Physiology) and extend a single dendrite to the epithelial border from where cilia radiate into the mucus that covers the olfactory epithelium. The cilia contain the signal transduction machinery that ultimately leads to excitatory current influx through the ciliary transduction channels, a cyclic nucleotide-gated (CNG) channel and a Ca2+-activated Cl- channel (Fig. 1). The ensuing depolarization triggers action potential generation at the cell body2-4.
In this video we describe the use of the "suction pipette technique" to record odorant-induced responses from ORNs. This method was originally developed to record from rod photoreceptors5 and a variant of this method can be found at jove.com modified to record from mouse cone photoreceptors6. The suction pipette technique was later adapted to also record from ORNs7,8. Briefly, following dissociation of the olfactory epithelium and cell isolation, the entire cell body of an ORN is sucked into the tip of a recording pipette. The dendrite and the cilia remain exposed to the bath solution and thus accessible to solution changes to enable e.g. odorant or pharmacological blocker application. In this configuration, no access to the intracellular environment is gained (no whole-cell voltage clamp) and the intracellular voltage remains free to vary. This allows the simultaneous recording of the slow receptor current that originates at the cilia and fast action potentials fired by the cell body9. The difference in kinetics between these two signals allows them to be separated using different filter settings. This technique can be used on any wild type or knockout mouse or to record selectively from ORNs that also express GFP to label specific subsets of ORNs, e.g. expressing a given odorant receptor or ion channel.

Olfactory receptor neurons (ORNs) convert odor signals first into a receptor current that in turn triggers action potentials that are conveyed to second order neurons in the olfactory bulb. Here we describe the suction pipette technique to record simultaneously the odorant-induced receptor current and action potentials from mouse ORNs.

从使用吸吸液管技术的嗅觉受体神经元的气味引起的反应记录

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral cortex. We here share our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex using a method developed by Lima and colleagues1. Recordings are performed in mice expressing Channelrhodopsin-2 (ChR2) in specific neuronal subpopulations. Members of the population are identified by their response to a brief flash of blue light. This technique &#8211; termed &#8220;PINP&#8221;, or Photostimulation-assisted Identification of Neuronal Populations &#8211; can be implemented with standard extracellular recording equipment. It can serve as an inexpensive and accessible alternative to calcium imaging or visually-guided patching, for the purpose of targeting extracellular recordings to genetically-identified cells. Here we provide a set of guidelines for optimizing the method in everyday practice. We refined our strategy specifically for targeting parvalbumin-positive (PV+) cells, but have found that it works for other interneuron types as well, such as somatostatin-expressing (SOM+) and calretinin-expressing (CR+) interneurons.

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

Here we describe our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex. Neurons expressing ChR2 are identified by their response to blue light. The method uses standard extracellular recording equipment, and serves as an inexpensive alternative to calcium imaging or visually-guided patching.

指南<em&gt;在体内</em&gt;单单元从Optogenetically鉴定皮质抑制性录像

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

A Free-breathing fMRI Method to Study Human Olfactory Function

The study of human olfaction is a highly complex and valuable field with applications ranging from biomedical research to clinical evaluation. Currently, evaluation of the functions of the human central olfactory system with functional magnetic resonance imaging (fMRI) is still a challenge because of several technical difficulties. There are some significant variables to take into account when considering an effective method for mapping the function of the central olfactory system using fMRI, including proper odorant selection, the interaction between odor presentation and respiration, and potential anticipation of or habituation to odorants. An event-related, respiration-triggered olfactory fMRI technique can accurately administer odorants to stimulate the olfactory system while minimizing potential interference. It can effectively capture the precise onsets of fMRI signals in the primary olfactory cortex using our data post-processing method. The technique presented here provides an efficient and practical means for generating reliable olfactory fMRI results. Such a technique can ultimately be applied in the clinical realm as a diagnostic tool for diseases associated with olfactory degeneration, including Alzheimer&#39;s and Parkinson&#39;s disease, as we begin to further understand the complexities of the human olfactory system.

We present the technical challenges and solutions for obtaining reliable functional magnetic resonance imaging (fMRI) data from the human central olfactory system. This includes special considerations in olfactory fMRI paradigm design, descriptions of fMRI data acquisition with an MRI-compatible olfactometer, odorant selection, and a special software tool for data post-processing.

一种免费呼吸功能磁共振成像方法研究人类嗅觉功能

The study of human olfaction is a highly complex and valuable field with applications ranging from biomedical research to clinical evaluation. Currently, ...

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a considerable amount of information has been generated regarding ORN responses to odorants, the role of specific ORNs in driving behavioral responses remains poorly understood. Complications in behavior analyses arise due to different volatilities of odorants that activate individual ORNs, multiple ORNs activated by single odorants, and the difficulty in replicating naturally observed temporal variations in olfactory stimuli using conventional odor-delivery methods in the laboratory. Here, we describe a protocol that analyzes Drosophila larval behavior in response to simultaneous optogenetic stimulation of its ORNs. The optogenetic technology used here allows for specificity of ORN activation and precise control of temporal patterns of ORN activation. Corresponding larval movement is tracked, digitally recorded, and analyzed using custom written software. By replacing odor stimuli with light stimuli, this method allows for a more precise control of individual ORN activation in order to study its impact on larval behavior. Our method could be further extended to study the impact of second-order projection neurons (PNs) as well as local neurons (LNs) on larval behavior. This method will thus enable a comprehensive dissection of olfactory circuit function and complement studies on how olfactory neuron activities translate in to behavior responses.

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

This protocol analyzes navigational behavior of Drosophila larva in response to simultaneous optogenetic stimulation of its olfactory neurons. Light of 630 nm wavelength is used to activate individual olfactory neurons expressing a red-shifted channel rhodopsin. Larval movement is simultaneously tracked, digitally recorded, and analyzed using custom-written software.

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

摘要

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此视频中的章节

在昆虫单感器记录果蝇按蚊

鼠标采用的空气相Electroolfactogram录制的嗅觉感官神经元响应分析

小龙虾的反思记录行为反应

使用方法比较物种调查父亲的反应神经生物学

从使用吸吸液管技术的嗅觉受体神经元的气味引起的反应记录

多单位记录方法来描述蝗虫的神经活动（

指南

为记录细胞内电压响应电法

一种免费呼吸功能磁共振成像方法研究人类嗅觉功能

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应