These studies were supported by grants from the National Institutes of Health, National Institute for Alcoholism and Alcohol Abuse to M.G. (P20AA017828, R01AA020634, P50 AA022537). The authors thank Jill Bettinger for helpful discussions and Jacqueline DeLoyht for technical assistance.

1天前分析<ol><li>在简短（1-5分钟） 二氧化碳收集苍蝇进入生鲜食品小瓶中的11（单一性别）组。 </li><li>允许蝇以回收的O / N的食品小瓶在环境受控的空间（通常为25℃，相对湿度60％，12小时光照/黑暗周期）。 </li><li>制备乙醇溶液（S）稀释在纯化（≥18MΩ）水纯（100％）乙醇至最终浓度（多个）适当的计划实验。允许溶液（S）返回RT O / N。 注意:乙醇稀释是放热的。 </li></ol>第2天的测定<ol><li>对于要测试的苍蝇的每个小瓶中，制备（ⅰ）一个干净的空的食品小瓶;即，测试小瓶中，（ii）一种新的醋酸纤维素插塞，（ⅲ）一个硅胶塞及（iv）将1ml乙醇溶液（ 见表3）。 </li><li>通过调节温度至20-25℃，相对湿度55-65％准备测试室。 </li><li>有另一名工人分配一个唯一的代码给每个组小瓶并记录代码购买。将编码小瓶苍蝇在试验室，以适应少数分钟。 </li><li>标签的空瓶子测试从2.3匹配飞瓶代码。 </li><li>通过以类似于表1中的电子表格输入码成列（1列/小瓶）构造一个硬拷贝测试日志。 </li><li>使用测试日志为指导，安排编码食品小瓶苍蝇和空测试小瓶入考场匹配阵列。 注:小瓶测试的合理的最大数量为24（ 即 6组，每组4瓶）。 </li><li>转让航班从食品到小瓶匹配/标记空测试小瓶，立即将醋酸纤维素插头插入测试小瓶，直到醋酸纤维素插头2厘米以下的小瓶顶部。 </li><li>此后，处理好四个小瓶每排为一组，在交错1分钟的间隔。 </li><li>对于时间0评估，单独瓦特掌握每个小瓶第i个拇指和食指，轻轻拍打桌子上三次敲苍蝇瓶的底部，等待30秒再算上苍蝇是不动/死亡人数。记录静止/死苍蝇为每小瓶的数量在时间0分钟的硬拷贝测试日志。 </li><li>启动定时器持续计数在时刻0，并立即开始加入1毫升乙醇到乙酸纤维素塞小瓶用于第一行/组4的小瓶。加入1毫升乙醇的醋酸纤维素插塞在小瓶以5秒的间隔在它们将被测试的顺序。加乙醇到乙酸纤维素塞的圆周运动，使乙醇在整个醋酸纤维素插塞均匀吸收。当乙醇已被添加到在该组的所有4测试小瓶，插入一个硅胶塞在每个小瓶密封。 </li><li>在时间1，2，3，4，5分钟，加入1毫升乙醇到第二，第三，第四，第五和第六组的4小瓶，分别。继续后插入硅胶瓶塞加入乙醇至每组4小瓶中。 </li><li>在时间6分钟，通过抓住每个小瓶用拇指和食指，轻轻拍打在桌子上三次敲苍蝇到小瓶的底部，等待30秒，然后计数并记录的总数测试所述第一组的4小瓶被麻醉苍蝇。得分苍蝇作为镇静剂，如果他们（我）站在瓶的地板，但不走或（ii）趴在自己的背上有或无拍打自己的翅膀。 </li><li>处理使用进度表2中的组在5秒的时间间隔内的每个小瓶中。 </li><li>在时间7，8，9，10和11分钟，测试所述第二，第三，第四，第五和第六组小瓶，分别作为完成对于第一组。 </li><li>在时间12分钟，再次测试所述第一组的4小瓶如在2.12中描述，并继续在13，14，15，16和17分钟，测试所述第二，第三，第四，第五和第六组的小瓶，分别。 </li><li>在2.12描述，直到所有的苍蝇有s继续进行测试飞行edated。 </li><li>在硬拷贝测试记录每个小瓶进入蝇的总数。从蝇的总数审查动/死苍蝇在时间0。 </li><li>计算在每个评估时间点的％活性苍蝇和情节数据作为​​％活性蝇（y轴）对时间（x轴）。通过从三阶多项式或S形曲线拟合内插镇静时间50的值（ST50，时间70％的镇静）或曲线下面积的计算定量乙醇的镇静。 </li><li>汇编从解码小瓶数据并执行统计分析（ 例如，单向ANOVA与Bonferroni多重比较检验）以适合的实验设计。 </li></ol>

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The authors have nothing to disclose.

简单的实验是重复定量有意义的表型是对行为的分析具有重要价值。这里所描述的工作涉及测量乙醇镇静敏感和快速耐受果蝇的实验的几个实际问题。虽然这项工作不是一个焦点，行为分析是通过维持对环境和遗传背景常数为测试对象的研究中变得容易。此外，比较通常应苍蝇饲养和测试并排侧的组之间进行比较。为此，从个人实验之内的所有的苍蝇在这项工作中有相同的遗传背景和均匀?...

酒精使用障碍（AUD）是一个巨大的健康问题的全球（1回顾）。虽然驱动澳元的发展机制是复杂的，这些疾病有重要的遗传成分（ 例如，2）。元和保守的行为反应的大遗传跨越许多物种（在3,4审查）已经产生在利用遗传模式生物，调查在乙醇相关的行为的特定基因的朝向更好地理解的分子基础的介入的强烈兴趣乙醇澳元。果蝇， 果蝇 ，已成为一个主要模式生物研究乙醇相关行为的分子遗传机制（3,4审查）。研究苍蝇突出角色的几个信号通路的行为反应，以乙醇（5审查）。有趣的是，某些基因和通路的影响行为respons下载到乙醇中蝇也被牵连在啮齿类乙醇相关的行为和/或人AUD（ 例如，6-14）。机制的保护驾驶乙醇相关的行为跨物种，加上在果蝇模型系统中可用的遗传工具套件，强调果蝇模型的效用调查的行为反应的遗传乙醇。 灵敏度15,16和公差（在17中综述）在人中乙醇被链接到元的发展。这两种行为反应的乙醇可在果蝇经由各种实验室测定法（在3,4综述）进行建模。所有已知的作者飞测定法是基于从乙醇镇静要么依赖于时间的乙醇诱导镇静/共济失调或依赖于时间的恢复。 从我们组对乙醇的灵敏度和r的遗传学以前的文章在果蝇 APID宽容的基础上，苍蝇乙醇蒸气引起镇静一个行为实验检测18。试验在该试验中被转印活成人发起蝇无需麻醉清空食品小瓶，诱捕苍蝇与醋酸纤维素插塞的小瓶中，加入乙醇到乙酸纤维素插塞的顶部（ 即，非飞边），并且密封含有苍蝇，醋酸纤维素插塞和乙醇用硅酮塞子的小瓶（参见示意图图S3中 ，参考18）。代表不同组的苍蝇的多个小瓶评估平行，提高吞吐量该测定的。小瓶给一个匿名代码和实验者被蒙蔽治疗组，以防止意外的偏见镇静的评估。在标准实验中，苍蝇在小瓶轻轻敲打以6分钟的间隔，并在30秒后恢复，镇静剂苍蝇在每个小瓶中的数目进行计数并转换器具d可％活性苍蝇。苍蝇吸收从醋酸纤维素插塞乙醇蒸汽以时间依赖的方式，导致逐步增加内部乙醇18和镇静（比照参考18和图1A和1B在本报告中）。镇静在此测定法操作上定义为果蝇（ⅰ）站立在没有步行或（ii）具有或不具有拍打翅膀躺在背上的。这里，这乙醇镇静测定进行详细说明，有关使用它进一步操作优化设置，并测定​​用于寻址的食物补充剂的选项上飞镇静灵敏度的贡献。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>food vials</td>
			<td>VWR</td>
			<td>89092-772</td>
			<td>narrow</td>
		</tr>
		<tr>
			<td>Flugs</td>
			<td>Genesee/flystuff.com</td>
			<td>49-102</td>
			<td>narrow</td>
		</tr>
		<tr>
			<td>silicone stopper</td>
			<td>Fisher Scientific</td>
			<td>09-704-1l</td>
			<td>#4</td>
		</tr>
		<tr>
			<td>ethanol</td>
			<td>Pharmaco-Aaper</td>
			<td>111000200</td>
			<td>200 proof</td>
		</tr>
	</tbody>
</table>

an inexpensive, scalable behavioral assay for measuring ethanol sedation sensitivity and rapid tolerance in drosophila

酒精使用障碍（AUD）是一个严重的健康挑战。尽管大量的遗传成分澳元，一些基因已被明确地牵连的病因。果蝇， 果蝇 ，是一个功能强大的模式探索潜在的酒精有关的行为的分子遗传机制，因此保持着识别和理解基因影响澳元的作用巨大潜力。利用果蝇模型对这些类型的研究依赖于测定中，能可靠地测量的行为反应，以乙醇的可用性。这份报告介绍了适用于评估乙醇的灵敏度和快速耐受在果蝇的实验。在该测定中测得的乙醇灵敏度由体积和乙醇浓度使用，各种先前报道遗传操作，并且时间还长的影响苍蝇收容没有食物之前立即测试。与此相反，乙醇sensitivity在此测定法测量不受苍蝇处理，苍蝇的性别，并补充以抗生素或活酵母生长培养基的活力。三种不同的方法来定量乙醇灵敏度描述，所有领先的基本上没有区别乙醇的敏感性结果。该测定的可伸缩的性质，结合其总体简单性设置和相对低的费用，使其适合于乙醇的灵敏度和快速耐受在果蝇的小和大规模基因分析。

从该乙醇的镇静测定的原始数据是蝇被镇静剂如乙醇蒸汽暴露时间的函数的数字。原始数据转换为％的活性苍蝇作为时间的函数（主要数据， 图1A，B，D - F）。灵敏度对乙醇的镇静从主数据可从曲线拟合来定量为镇静时间50（ST50），成为曲线（AUC）下的镇静剂或AEA所需苍蝇的50％的时间，通过内插。先前报道18三阶多项式曲线拟合的主数据以及预期的（平均，R <sup...

Watch this Scientific Journal Video about An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila at JoVE.com

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

一种廉价的，可扩展的行为分析测量乙醇镇静灵敏度和快速耐受<em&gt;果蝇</em

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in ...

an-inexpensive-scalable-behavioral-assay-for-measuring-ethanol

Research

JoVE Journal

Neuroscience

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

10.2K 次观看.  Virginia Commonwealth University. Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

视频: An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

睾酮和社会决策，结合行为内分泌和实验经济学:

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

科研

神经科学

A Simple Way to Measure Ethanol Sensitivity in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain is correlated with that strain s ethanol preference, and therefore sedation is a highly relevant measure to study the genetics of alcohol responses and drinking. We demonstrate a simple way to expose flies to ethanol and measure its intoxicating effects. The assay we describe can determine acute sensitivity, as well as ethanol tolerance induced by repeat exposure. It does not require a technically involved setup, and can therefore be applied in any laboratory with basic fly culture tools.

A simple assay to measure the sedating effects of ethanol on Drosophila flies, based on the loss of righting reflex, is described.

一个简单的方法来衡量在果蝇的乙醇灵敏度

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

评估大鼠蔗糖渴求育成的一个通用方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

测量乙醇的影响上的步态速度的测定<em&gt;秀丽隐杆线虫</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.

一个行为分析对机械感觉MARCM为基础的克隆<em&gt;果蝇</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

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 ...

Measuring and Altering Mating Drive in Male Drosophila melanogaster

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, reductionist, and scalable system for in-depth investigation of motivation using the mating drive of male Drosophila&#160;melanogaster (Drosophila), the methods for which we detail here. The behavioral paradigm centers on the finding that male mating drive decreases alongside fertility over the course of repeated copulations and recovers over ~3 d. In this system, the powerful neurogenetic tools available in the fly converge with the genetic accessibility and putative wiring diagram available for sexual behavior. This convergence allows rapid isolation and interrogation of small neuronal populations with specific motivational functions. Here we detail the design and execution of the satiety assay that is used to measure and alter courtship motivation in the male fly. Using this assay, we also demonstrate that low male mating drive can be overcome by stimulating dopaminergic neurons. The satiety assay is simple, affordable, and robust to influences of genetic background. We expect the satiety assay to generate many new insights into the neurobiology of motivational states.

Measuring and Altering Mating Drive in Male Drosophila melanogaster

This article describes a behavioral assay that uses male mating drive in Drosophila melanogaster to study motivation. Using this method, researchers can utilize advanced fly neurogenetic techniques to uncover the genetic, molecular, and cellular mechanisms that underlie this motivation.

测量并改变配套驱动马累<em&gt;果蝇</em

Despite decades of investigation, the neuronal and molecular bases of motivational states remain mysterious. We have recently developed a novel, ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.