我们感谢阿拉文森·塞缪尔博士和马克·格肖博士帮助开发了行为装置的初始版本，马修·巴蒂博士修改了行为装置。这项研究得到了辛辛那提儿童医院、JST/PRESTO、迪姆斯和NIH R01 GM107582到F.N.H.的受托人格兰特的支持。

1. 准备苍蝇
<ol><li>浅暗（LD）实验
	<ol>
<li>在光12小时/暗12小时（LD）周期下，在孵化器中提高苍蝇（25°C/40-60%相对湿度（RH））。孵化器的光强度为 500-1，000 勒克斯。</li>
		<li>在 24 小时内完成行为检测需要两个孵化器。两个孵化器都应具有具有开机功能的可编程光。它们还应有无法透射到光的实心门（即 没有玻璃或玻璃杯）。 注意:一个孵化器应指定为"日"孵化器，并设置为 LD 周期，12 小时光和 12 小时黑暗。第二个孵化器应指定为"夜间"孵化器，并设置为第一个孵化器的反向，12小时黑暗，然后是12小时光。夜间孵化器必须放置在黑暗中可访问的房间里，这样，体验夜间条件的苍蝇就可以轻松地进入实验。</li>
		<li>将飞瓶放在白天或晚上的孵化器中。收集新孵化的苍蝇在一个新的小瓶，20-30每检测，并保持在同一孵化器2-3天。</li>
		<li>2-3天后，使用苍蝇进行温度偏好行为检测。<ol>
<li>对于日（泽格伯时间 （ZT） 0-12） 实验，从日孵化器收集苍蝇。</li>
			<li>就在行为实验之前，将收集的飞瓶带出日间孵化器。</li>
			<li>对于夜间（ZT 13-24）实验，从夜间孵化器收集苍蝇。</li>
			<li>就在行为实验之前，将收集的飞瓶带出夜间孵化器，用铝箔包裹，放在暗室的一个盒子里，放在红灯下。 注意:由于夜间实验的温度偏好行为检测是在黑暗中进行的，所以必须防止对苍蝇的光线暴露，直到行为实验结束。 注意:在实验进行当天，苍蝇不应接触到二氧化碳。</li>
		</ol>
</li>
	</ol></li>
	<li>恒定黑暗 （DD） 实验
	<ol>
<li>
DD日
		<ol>
<li>DD 日实验需要额外的孵化器，我们称之为手稿其余部分的"过渡"孵化器。过渡孵化器必须放置在黑暗中可访问的房间里，这样，体验DD条件的苍蝇就可以轻松访问进行实验。过渡孵化器的示例灯时间表将在下午 1 点到 7 点开灯，晚上 7 点到 1 点（图 1）关闭。</li>
			<li>收集在日孵化器中饲养的苍蝇。当灯亮起时，在下午 1 点到 7 点之间将飞瓶放在过渡孵化器中。这样，苍蝇被正确地暴露在光线下，直到晚上7点，此时光线关闭。</li>
			<li>第二天下午1点前，在黑暗条件下，将飞瓶从过渡孵化器中取出，用铝箔包裹，放在一个盒子里。将盒子放在任何孵化器中再放一天。</li>
		</ol>
</li>
		<li>
DD之夜
		<ol>
<li>收集在暗黑中在夜孵化器中升起的苍蝇，在红灯下。或在夜间孵化器的灯光亮起时额外收集苍蝇。</li>
			<li>只要灯熄灭，在黑暗中用铝箔包裹收集的小瓶，并将小瓶放在一个盒子里。将盒子放在任何孵化器中再放两天（图1B）。 注意:由于夜间实验的温度偏好行为检测是在黑暗中进行的，所以必须防止对苍蝇的光线暴露，直到行为实验结束。</li>
		</ol>
</li>
	</ol></li>
	<li>恒光（LL）实验
	<ol>
<li>
LL 日
		<ol>
<li>LL日实验需要额外的孵化器。此孵化器保持LL条件（25°C，800勒克斯），并持续开灯。</li>
			<li>收集在日培养箱中饲养的苍蝇。在其"白天"随时将飞瓶放在LL孵化器中。</li>
		</ol>
</li>
		<li>
LL 之夜
		<ol>
<li>收集在夜间孵化器中饲养的苍蝇。在夜间孵化器的灯光打开期间，将飞瓶从夜间孵化器转移到LL孵化器。 注意:例如，夜间孵化器中的灯在上午 7 点关闭。早上 7 点前将飞瓶从夜间孵化器转移到 LL 孵化器，并将小瓶在 LL 孵化器中再保存 4 天。 注:在LL条件下的第4天，运动活动的振荡被废除15，16，而TPR仍然持续3。</li>
		</ol>
</li>
	</ol></li>
</ol>2. 温度偏好行为测定装置
<ol><li>将玻璃杯盖（29 厘米 x 19.2 厘米）（图 4）放在铝板上。</li>
	<li>监测板与盖之间的温度。六个温度探针在一条车道内盖内的不同位置连接（图2）。 注意:确保探针不接触铝板或玻璃杯盖。气温应设置为 18-32 °C 的梯度。</li>
	<li>将设备放置在维持在 25 °C/65-75% RH 的环境室中。这个房间需要被封锁，远离任何外部光线。环境室通常配有风扇，以保持一定的温度和湿度。 注意:风扇的空气可能会干扰仪器上稳定的温度梯度。为了防止这种情况，我们使用透明纸张，覆盖仪器周围的区域。</li>
	<li>准备温度计和催眠仪，检查环境室的温度和湿度。 注意:光影响 嗜血杆菌3的温度偏好。设备上应均匀提供相同强度的光。我们的环境室灯的强度是~800勒克斯。 注意:完成行为实验后，将从 CO2 罐连接的管子或电源放在仪器顶部孔附近，以便麻醉和清除苍蝇。</li>
</ol>3. 准备使用设备
<ol><li>打开仪器至少30分钟，以便正确确定板表面的温度梯度（图3）。</li>
	<li>将行为装置的盖子涂上防水剂，以防止苍蝇攀爬盖子的墙壁或天花板。擦去多余的防水剂，将盖子保持25-30分钟干燥。</li>
	<li>去除铝板上的任何凝结物。将玻璃杯盖放在铝板上，用六个 C 夹子固定（图 2）。 注意:如果有必要，盖子密封良好是非常重要的。</li>
	<li>将盖子保留至少 15 分钟。铝板和盖子之间的空气温度梯度为 18-32 °C。</li>
</ol>4. 温度偏好行为测定
<ol><li>通过盖子每个车道中心的小孔（图 2 和 4）将苍蝇装载到铝板和仪器的玻璃盖之间的空间。用盖子遮盖孔，防止苍蝇逃跑。</li>
	<li>对于黑暗的环境，关掉环境室的所有灯。当苍蝇被放入仪器时，可以使用红灯。在行为实验完成之前，确保苍蝇不会暴露在除红灯以外的任何灯光下。</li>
	<li>对于每次试验，使用20-25只成年苍蝇，这些苍蝇不会在随后的试验中重复使用。行为检测执行 30 分钟。拍几张照片有或没有闪光灯。在实验过程中，要小心不要发出太大的噪音或任何突然的动作。</li>
	<li>记录仪器上所有六个探针的温度。注意室温和湿度。</li>
	<li>用二氧化碳气体麻醉仪器中的苍蝇，松开夹子，取下玻璃杯盖，将苍蝇从盘子中取出。每次实验后，苍蝇都会被丢弃。擦去盘子里的任何凝结物或水分。更换板上的盖子，用夹子拧紧，为下一个实验做准备。</li>
	<li>为了代表全天的温度偏好，24 小时时段分为 8 个时区，白天 4 个时区，晚上 4 个时区。例如，我们使用这些 ZT 或 CT 1-3、4-6、7-9、10-12、13-15、16-18、19-21 和 22-24。 注意:由于遮蔽效果引起的表型变化预计在开启 （ZT0） 或关闭 （ZT12） 之后立即发生，我们不检查这些时间的温度偏好行为（ZT 或 CT 0-1、11.5-13 和 23.5-24）。每个时区必须至少进行五次试验，以便结果在统计学上是合理的。</li>
</ol>5. 数据分析
<ol><li>计算温度梯度如下:根据放置在沿边缘的玻璃盖顶部和底部的两个尺子（图 2A）确定温度探针的位置。</li>
	<li>温度探针之间的温度梯度估计是线性的。根据温度探头的位置及其相应的记录温度，在图片的适当位置绘制代表每个温度的线条。计算位于每个度区间的苍蝇数量。不包括墙壁上的任何苍蝇或盖子的天花板。</li>
	<li>计算每个车道每个温度范围内的苍蝇百分比。平均首选温度是通过计算每个间隔的苍蝇和温度百分比的产品计算的，如下所示: 百分比 的苍蝇 x 18.5 °C = 百分比的苍蝇 x 19.5 °C = 百分比的苍蝇 x 20.5 °C 。。</li>
	<li>计算每个时区的平均首选温度:每个时区（ZT 1-3、4-6、7-9、10-12、13-15、16-18、19-21 和 22-24）执行温度偏好行为>5 次。要计算每个时区的平均首选温度，每个试验的平均首选温度一起平均。s.e..m错误栏等于试验之间的错误。</li>
</ol>

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没有什么可透露的。

在这里，我们说明了温度偏好行为装置的细节和TPR行为分析。 德罗索菲拉 展示了时钟控制的 TPR 的突出、坚固和可重复的功能。然而，我们的数据表明，至少有两个因素，环境光和年龄，严重扰乱TPR行为表型。
我们观察到，光显著影响 德罗索菲拉的温度偏好。这与在LD中保存 的1118 只苍蝇比DD中的苍蝇更喜欢白天的温度更高，尽管在LD和DD3</sup...

温度是一种无处不在的环境线索。动物表现出各种行为，以避免有害的温度和寻求舒适的。 嗜血杆菌 表现出一个强大的温度偏好行为6，7。当苍蝇被释放到温度梯度从18-32 °C，苍蝇避免温暖和寒冷的温度，最后选择在3日早晨25°C的首选温度。暖温传感器是一组热感官神经元，交流神经元，它表达 嗜血杆菌 瞬态受体电位（TPR）通道，TRPA16，9。冷温传感器位于第三天线段，因为消融第三个天线段会导致缺乏冷温避免6。最近，TRPP蛋白布里维多（Brv）被确定为10。由于 Brv 在第三个天线段中表示并调解冷检测，Brv 是一种可能的冷感应分子，对温度偏好行为至关重要。总之，苍蝇使用这两个温度传感器来避免温暖和寒冷的温度，并找到一个首选的温度。
虽然哺乳动物产生热量来调节体温，但息热通常使体温适应环境温度11。众所周知，一些教会每天都会表现出一种TPR行为，这被认为是教会调节BTR12的策略。为了确定苍蝇是否表现出TPR，我们在24小时内重复了不同点的温度偏好行为分析。我们发现， 德罗索菲拉 每天表现出TPR，这是在早上低，在晚上高，并遵循类似于BTR在人类13的模式。
在 德罗索菲拉，大脑中有大约150个时钟神经元。调节运动活动的时钟神经元称为M和E振荡器。然而，有趣的是，M和E振荡器不调节TPR，相反，我们表明，DN2时钟神经元在大脑中调节TPR，但不是运动活动。这些数据表明，TPR独立于运动活动进行调节。值得注意的是，哺乳动物BTR也独立于运动活动调节。大鼠的消融研究表明，BTR通过特定的SCN神经元进行控制，这些神经元针对的是与控制运动活动14的神经元不同的亚参数区神经元。因此，我们的数据考虑了哺乳动物BTR和苍蝇TPR进化保存的可能性3，因为飞TPR和哺乳动物BTR都表现出昼夜时钟依赖温度的节奏，这是独立于运动运动活动的调节。
在这里，我们描述了如何分析 德罗索菲拉的TPR行为检测的细节。这种方法不仅可以研究TPR的分子机制和神经回路，还可以研究大脑如何整合不同的环境线索和内部生物钟。

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design and analysis of temperature preference behavior and its circadian rhythm in drosophila

昼夜钟调节生活的许多方面，包括睡眠，运动活动，和体温（BTR）节律1，2。我们最近发现了一种新的环食素输出，称为温度偏好节奏（TPR），其中苍蝇的首选温度在白天上升，在夜间下降3。令人惊讶的是，TPR和运动活性通过不同的昼夜神经元3控制。恐龙运动活动是一种众所周知的昼夜行为输出，为发现许多保存的哺乳动物昼夜钟基因和机制4提供了强有力的贡献。因此，了解TPR将导致迄今未知的分子和细胞昼夜机制的识别。在这里，我们描述如何执行和分析 TPR 分析。这项技术不仅能够解剖TPR的分子和神经机制，而且为整合不同环境信号和调节动物行为的大脑功能的基本机制提供了新的见解。此外，我们最近公布的数据表明，苍蝇TPR与哺乳动物BTR3共享功能。嗜血杆菌是息息的，其中通常调节体温的行为。因此，TPR是一种策略，用于在这些苍蝇5-8产生有节奏的体温。我们相信，进一步探索果蝇TPR将有助于描述动物体温控制机制的特征。

温度偏好节奏的示例显示在图5 中。如果行为程序成功，苍蝇应表现出TPR，在TPR中，它们更喜欢在早晨的低温和在晚上的较高温度。无论遗传背景如何，白天温度偏好的+1-1.5 °C增加应在白天观察到，因为我们显示，w1118、yw和广州 S 苍蝇在白天3中表现出类似的温度偏好。
<img alt="Figure 1" fo:content-width=...

Watch this Scientific Journal Video about Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila at JoVE.com

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

我们最近发现了一种新的 环食 素输出，温度偏好节奏（TPR），其中苍蝇的首选温度在白天上升，在夜间下降。TPR 独立于另一个昼夜输出（运动活动）进行调节。在这里，我们描述了在 德罗索菲拉TPR的设计和分析。

温度偏好行为及其在德罗索菲拉的昼夜节律的设计与分析

The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a ...

design-analysis-temperature-preference-behavior-its-circadian-rhythm

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Cell Biology

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Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

Unit 1

Alternative Signaling Routes in Gene Expression

SCIENTISTS IN ACTION

8.0K 次观看.  Cincinnati Childrens Hospital Medical Center. 我们最近发现了一种新的 环食 素输出，温度偏好节奏（TPR），其中苍蝇的首选温度在白天上升，在夜间下降。TPR 独立于另一个昼夜输出（运动活动）进行调节。在这里，我们描述了在 德罗索菲拉TPR的设计和分析。

视频: Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

含量自发活动，研究昼夜节律和睡眠参数果蝇</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

科研

神经科学

A Computational Method to Quantify Fly Circadian Activity

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. Fundamental mechanisms that underlie this temporal control are widely studied using the fruit fly Drosophila melanogaster as a model organism. In flies, the clock is typically studied by analyzing multiday locomotor recording. Such a recording shows a complex bimodal pattern with two peaks of activity: a morning peak that happens around dawn, and an evening peak that happens around dusk. These two peaks together form a waveform that is very different from sinusoidal oscillations observed in clock genes, suggesting that mechanisms in addition to the clock have profound effects in producing the observed patterns in behavioral data. Here we provide instructions on using a recently developed computational method that mathematically describes temporal patterns in fly activity. The method fits activity data with a model waveform that consists of four exponential terms and nine independent parameters that fully describe the shape and size of the morning and evening peaks of activity. The extracted parameters can help elucidate the kinetic mechanisms of substrates that underlie the commonly observed bimodal activity patterns in fly locomotor rhythms.

A method to quantify the main temporal features seen in fly circadian locomotor rhythms is presented. The quantification is achieved by fitting fly activity with a multi-parametric model waveform. The model parameters describe the shape and size of the morning and evening peaks of daily activity.

量化飞行昼夜活动的一种计算方法

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. ...

行为学

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific responses to changing temperatures according to their physiological tolerance and adaptability. Drosophila flies also possess a temperature sensing system that has become fundamental to understanding the neural basis of temperature processing in ectotherms. We present here a temperature-controlled arena that permits fast and precise temperature changes with temporal and spatial control to explore the response of individual flies to changing temperatures. Individual flies are placed in the arena and exposed to pre-programmed temperature challenges, such as uniform gradual increases in temperature to determine reaction norms or spatially distributed temperatures at the same time to determine preferences. Individuals are automatically tracked, allowing the quantification of speed or location preference. This method can be used to rapidly quantify the response over a large range of temperatures to determine temperature performance curves in Drosophila or other insects of similar size. In addition, it can be used for genetic studies to quantify temperature preferences and reactions of mutants or wild-type flies. This method can help uncover the basis of thermal speciation and adaptation, as well as the neural mechanisms behind temperature processing.

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Here we present a protocol to automatically determine the locomotor performance of Drosophila at changing temperatures using a programmable temperature-controlled arena that produces fast and accurate temperature changes in time and space.

一种自动测定果蝇在空间和时间温度变化中的性能的方法

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific ...

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer basic questions on how light information is processed and shared between rapid and circadian behaviors. Drosophila larvae display a stereotypical avoidance behavior when exposed to light. To investigate light dependent behaviors comparably simple light-dark preference tests can be applied. In vertebrates and arthropods the neural pathways involved in sensing and processing visual inputs partially overlap with those processing photic circadian information. The fascinating question of how the light sensing system and the circadian system interact to keep behavioral outputs coordinated remains largely unexplored. Drosophila is an impacting biological model to approach these questions, due to a small number of neurons in the brain and the availability of genetic tools for neuronal manipulation. The presented light-dark preference assay allows the investigation of a range of visual behaviors including circadian control of phototaxis.

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Here we describe a light-dark preference test for Drosophila larva. This assay provides information about innate and circadian regulation of light sensing and processing photobehavior.

光偏好的检测，以研究的先天和昼夜调节感光特性<em&gt;果蝇</em&gt;幼虫

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer ...

Conditions Affecting Social Space in Drosophila melanogaster

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a straightforward manner. As we previously demonstrated 1, in a two-dimensional chamber, we first force the flies to form a tight group, subsequently allowing them to take their preferred distance from each other. After the flies have settled, we measure the distance to the closest neighbor (or social space), processing a static picture with free online software (ImageJ). The analysis of the distance to the closest neighbor allows researchers to determine the effects of genetic and environmental factors on social interaction, while controlling for potential confounding factors. Diverse factors such as climbing ability, time of day, sex, and number of flies, can modify social spacing of flies. We thus propose a series of experimental controls to mitigate these confounding effects. This assay can be used for at least two purposes. First, researchers can determine how their favorite environmental shift (such as isolation, temperature, stress or toxins) will impact social spacing 1,2. Second, researchers can dissect the genetic and neural underpinnings of this basic form of social behavior 1,3. Specifically, we used it as a diagnostic tool to study the role of orthologous genes thought to be involved in social behavior in other organisms, such as candidate genes for autism in humans 4.

Conditions Affecting Social Space in Drosophila melanogaster

The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm.

在条件影响社会空间<em&gt;果蝇</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae

Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences in&#160;environmental temperature, which enables them to seek out their preferred thermal landscape. To define the temperature preferences of larvae over a defined linear range, we developed an assay using a temperature gradient. To establish a single-directional gradient, two aluminum blocks are connected to independent water baths, each of which controls the temperature of individual blocks. The two blocks set the lower and upper limits of the gradient. The temperature gradient is established by placing an agarose-coated aluminum plate over the two water-controlled blocks so that the plate spans the distance between them. The ends of the aluminum plate that is set on the top of the water blocks defines the minimum and maximum temperatures, and the regions in-between the two blocks form a linear temperature gradient. The gradient assay can be applied to&#160;larvae of different ages and can be used to identify mutants that exhibit phenotypes, such as those with mutations affecting genes encoding TRP channels and opsins, which are required for temperature discrimination.

A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae

Here, we present a protocol to determine the preferred environmental temperature of Drosophila larvae using a continuous thermal gradient.

温度梯度法测定果蝇幼虫的热偏好

Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences in&#160;environmental temperature, which ...

Analysis of Circadian Photoresponses in Drosophila Using Locomotor Activity

Circadian rhythms are only beneficial to animals if they can be synchronized by changes of ambient conditions. Light and temperature are two dominant environmental parameters that synchronize animal circadian clocks. In Drosophila circadian photo-responses are mediated by a solely blue light photoreceptor named CRYPTOCHROME (CRY). Upon photoreception, CRY changes its conformation and initiates the proteosomal dependent degradation of TIMELESS (TIM). TIM is an important pacemaker protein, thus degradation of TIM will reset the circadian clock. Under constant light conditions (LL), wild type flies quickly become arrhythmic because of the constant degradation of TIM, while flies bearing defects with circadian photo-responses will still be rhythmic. Thus LL triggered arrhythmicity has been used for screening of components in circadian light input pathways. A brief short light pulse in the night can also dramatically shift phases of circadian rhythms. As expected, this phase shift response is reduced in flied with defects in circadian photoresponse. Thus analyzing locomotion behavior rhythmicity under LL or phase changes after short light pulse in constant darkness (DD) are two major methods to study circadian photoresponse. Here we describe how to design and analyze LL and phase response experiments. LL arrhythmicity is suitable for screening light input pathways mutants, whereas phase response validates the results and provide further information for light sensitivity.

Drosophila locomotor activity is a robust and quantitative measurement of circadian photo-responses. We describe protocols for designing behavior experiments for circadian photo-responses and analyzing the data. Studying the circadian photo-responses is important for dissecting the neuronal and molecular mechanisms of light entrainment.

Circadian rhythms are only beneficial to animals if they can be synchronized by changes of ambient conditions. Light and temperature are two dominant ...

生物学

Circadian Entrainment of Drosophila Melanogaster

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this rhythm and to understand the resulting outputs, entrainment of model organisms to defined circadian time-points is required. Here we detail a procedure to entrain many Drosophila to a defined circadian rhythm. Furthermore, we detail post-entrainment steps to prepare samples for immunofluorescence, nucleic acid, or protein extraction-based analysis.

Here, we detail how to synchronize Drosophila to a circadian day. This is the first, and most important step necessary for studying biological rhythms and chronobiology.

果蝇黑色素的循环训练

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this ...

Determining Temperature Preference of Mosquitoes and Other Ectotherms

Most insects and other ectotherms have a relatively narrow optimal temperature window, and deviation from their optima can have significant effects on their fitness, as well as other characteristics. Consequently, many such ectotherms seek out their optimal temperature range. Although temperature preferences of mosquitoes and other insects have been well studied, the traditional experimental setup is performed using a temperature gradient on an aluminum surface in a highly enclosed space. In some cases, this equipment restricts many natural behaviors, such as flying, which may be important in preference selection.
The objective of this study is to observe insect preference for air temperature by using a two-chamber apparatus with sufficient room for flight. The two chambers consist of independent temperature-controlled incubators, each with a large aperture. The incubators are connected by these apertures using a short acrylic bridge. Inside the incubators are two netted cages, linked via the apertures and bridge, allowing the insects to freely fly between the different conditions. The acrylic bridge also acts as a temperature gradient between the two incubators.
Due to the spacious area in the cage and easy construction, this method can be used to study any small ectotherm and/or any manipulation which may alter temperature preference including sensory organ manipulation, diet, gut flora, and endosymbiont presence at biosafety levels 1 or 2 (BSL 1 or 2). Additionally, the apparatus can be used for the study of pathogen infection using further containment (e.g., inside of a biosafety cabinet) at BSL 3.

Insects have an optimal environmental temperature range which they seek to remain within, and many external and internal factors can alter this preference. Here, we describe a cost-effective and simple method to study temperature choice, which allows insects to freely exhibit their natural behaviors.

确定蚊子和其他变温动物的温度偏好

Most insects and other ectotherms have a relatively narrow optimal temperature window, and deviation from their optima can have significant effects on ...

Temperature Gradient Assay: A Method to Test Temperature Preference in Drosophila Larvae

Source: Liu, J., et al. <a href="https://www.jove.com/video/57963/" target="_blank">A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae</a>. J. Vis. Exp. (2018).
Drosophila fruit flies can distinguish small changes in temperature and therefore select environments with favorable thermal conditions. This video describes a behavioral assay that tests the temperature preference of Drosophila larvae on a linear thermal gradient.

温度梯度测定:一种测试果蝇幼虫温度偏好的方法

Source: Liu, J., et al. A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae. J. Vis. Exp. (2018).
Drosophila fruit flies ...

温度偏好行为及其在德罗索菲拉的昼夜节律的设计与分析

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含量自发活动，研究昼夜节律和睡眠参数果蝇