Name: functional analysis of the larval feeding circuit in drosophila
Uploaded: 2013-11-19T16:45:00
Description: Watch this Scientific Journal Video about Functional Analysis of the Larval Feeding Circuit in Drosophila at JoVE.com

作者要感谢总统的研究基金来自圣路易斯大学授予的WSN

1。人口笼的保养<ol><li>保持人口笼在25°C在12小时明暗周期。只要在对照组和实验组暴露在相同的照明条件，那么这种技术可以在一个标准的实验室设置进行。 </li><li>让女性在一夜之间产卵苹果汁琼脂平皿上。 </li><li>通过保持板与新蛋存放在25℃，24小时收集初孵幼虫。放置一小团酵母中的板的中心，以吸引孵出幼虫。 </li><li>收集已迁移到酵母膏苹果汁板中心第一龄幼虫。使用金​​属...

<ol>
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血清素stomatogastric电路，它在胚胎发育晚期出现的异常发育，将影响其成熟的功能。变化突起架构支配肠道可与电路的功能输出，这是进料速率（通过嘴钩收缩在酵母溶液测定）（图1）相关联。在果蝇中使用UAS-Gal4的二分体系使得可以特异性靶向上调或下调表达一个给定的转录物至特定组织的;改变一个给定的蛋白质的表达，可以精确地定量给予适当的工具。这种技术可以用来?...

果蝇是研究神经回路的发展，由于快速的生成时间，较低的实验成本，操纵和控制遗传和环境因素的能力是非常强大的模型系统。神经再生，神经元路径的发现和突触是人类和果蝇中保守，因此在创建，维护和修改神经回路机制是保守为好。 经典的神经递质，例如血清素（5 -羟色胺，或5-HT）可以作为生长因子通过它们的作用如同在成熟的神经电路1-3的信号分子前 以往的研究表明，扰动的5-HT水平胚胎发育过程中改变成熟神经元4的连通性。其他人已经表明5 -羟色胺对培养的神经元Helisoma异位应用抑制神经突生长以及突触5-7。 在 D中rosophila，发育的5-HT水平负相关静脉曲张数量和大小，以及aborization的程度，沿着突起从中枢神经系统8投射到前肠的长度。 血清素能神经传递已经显示调节摄食行为在不同的物种，包括果蝇 8-9。馈电电路在果蝇是可以作为一个模型关联功能的输出（馈送）与在轴突突起从大脑前肠的发展改变一个相对简单的电路。 Schoofs 等 。已经表明， 果蝇幼虫取食是由中枢模式发生器，影响肌肉组织10规范。而特定的肌肉解剖尚未完全了解，但已经表明，触角神经，上颌神经和前胸腺副神经负责参与肌肉目标摄食行为。涉及的无脊椎动物的摄食肌肉和神经解剖大多数数据是有限的丽蝇幼虫。 第二龄期幼虫的进料速率可以通过cephalopharyngeal骨片（口钩）的回缩进行评估，并且是可重复的和高吞吐量。该cephalopharyngeal板由中枢5-HT能神经元纤维通过额叶神经支配。腺胃，或前肠，是由血清素纤维（recurrens神经），该束状中肠，并负责前肠的收缩（ 图1）11-12支配。变化的轴突分枝和静脉曲张沿神经突长度的数量和大小，可以用免疫组织化学技术进行定量。在开发过程中操纵神经元的5-HT，无论是直接或间接地，可改变该供电电路的功能输出，其可以进行评估，并以变化的morpholo相关GY的突起结构。

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			<td height="40" style="height:40px;width:217px;">Eclipse E-800 Microscope</td>
			<td style="width:175px;">Nikon Instruments</td>
			<td style="width:115px;"></td>
			<td style="width:251px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:217px;">Neuroleucida</td>
			<td style="width:175px;">MBF Biosciences</td>
			<td style="width:115px;">NL-15</td>
			<td style="width:251px;">Used to analyze gut fiber architecture, not necessary to have</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:217px;">Northern Eclipse</td>
			<td style="width:175px;">Empix Inc</td>
			<td style="width:115px;"></td>
			<td style="width:251px;">Imaging software</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:217px;">G-2E/C TRITC EX 528-553</td>
			<td style="width:175px;">Nikon Instruments</td>
			<td style="width:115px;">96312</td>
			<td style="width:251px;">Filter for specific secondary antibody</td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:217px;">N.A. 0.75; W.D. 0.72 mm; DIC Prism: 40xI, 40x I-C; Spring loaded</td>
			<td style="width:175px;">Nikon Instruments</td>
			<td style="width:115px;">MRH00400</td>
			<td style="width:251px;">Objective used for imaging</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:217px;">Simple Neurite Tracer</td>
			<td style="width:175px;">NIH Image J</td>
			<td style="width:115px;"></td>
			<td style="width:251px;"><a href="http://fiji.sc/Simple_Neurite_Tracer">http://fiji.sc/Simple_Neurite_Tracer</a></td>
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functional analysis of the larval feeding circuit in drosophila

的血清素激活的供电电路在果蝇幼虫可用于该电路的开发过程中，调查至关重要的神经基质。使用该电路的功能输出，馈送，改变stomatogastric系统的神经细胞的体系结构可以被可视化。摄食行为可以通过观察口的钩子，它从大脑接收支配收缩的速率进行记录。自发行为作为一种生理控制觅食，因为幼虫用自己的嘴钩来遍历整个琼脂基质。改变摄食行为可以与突起的轴突架构相关联支配的内脏。采用免疫组织化学也能够可视化和定量这些变化。幼虫在行为范式操作不当可能会改变数据，因为他们是操纵非常敏感。适当的成像轴突架构的支配肠道是膨体的数量和尺寸的精确定量以及分支节点的程度是至关重要的。大多数电路的分析只允许对神经突的体系结构或行为效果的视觉效果，但是，这种模型允许一个相关联的电路与在神经元体系结构的损伤的功能性的输出。

血清素供电电路中的果蝇幼虫可以作为一个非常有效的模型，观察特定因素对神经系统发育的影响。通过定量进料速率，因此能够与它的功能性输出（图1）连接的馈电电路的轴突的结构。的运动功能测定法是用来作为一种生理控制的口钩的后退动作中，由于幼虫用他们的嘴钩穿过琼脂表面推动自己。应在控制和突变基因型之间的自发反应没有区别，如果突变只影响供电电路<str...

Watch this Scientific Journal Video about Functional Analysis of the Larval Feeding Circuit in Drosophila at JoVE.com

Functional Analysis of the Larval Feeding Circuit in Drosophila

供电电路在果蝇幼虫提供了一个简单但功能强大的模型，它允许改变喂养率与在stomatogastric神经回路的改变相关。这个电路是由该发送突起的钩口以及前肠中央血清素能神经元。

在幼虫馈电电路的功能分析果蝇</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

Research

JoVE Journal

Neuroscience

Functional Analysis of the Larval Feeding Circuit in Drosophila

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

3幼虫节段性神经的可视化 RD龄果蝇幼虫的筹备工作

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

科研

神经科学

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

在果蝇幼虫分部轴突在体内的突触小泡的可视化

Elucidating the mechanisms of axonal transport has shown to be very important in determining how defects in long distance transport affect different ...

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

形态分析果蝇幼虫外周感觉神经元树突和轴突使用遗传马赛克

Nervous system development requires the correct specification of neuron position and identity, followed by accurate neuron class-specific dendritic ...

Visualization of Proprioceptors in Drosophila Larvae and Pupae

Visualization of Proprioceptors in Drosophila Larvae and Pupae

可视化本体感受器<em&gt;果蝇</em&gt;幼虫和蛹

Proprioception is the ability to sense the motion, or position, of body parts by responding to stimuli arising within the body. In fruitflies and other ...

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

幼虫，蛹和成人的解剖和免疫组化<em&gt;果蝇</em&gt;视网膜

The compound eye of Drosophila melanogaster consists of about 750 ommatidia (unit eyes). Each ommatidium is composed of about 20 cells, including ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

嗅觉受体神经元在全山免疫标记<em&gt;果蝇</em&gt;天线

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Live Imaging of Drosophila Larval Neuroblasts

Live Imaging of Drosophila Larval Neuroblasts

的实时成像<em&gt;果蝇</em&gt;幼虫神经母细胞

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

为什么量化事项：表型的表征在<em&gt;果蝇</em&gt;幼虫神经肌肉接头

Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic ...

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

去除<em&gt;果蝇</em从幼虫鱼片的感官神经元和表皮细胞的免疫荧光分析&gt;肌肉组织

Drosophila larval dendritic arborization (da) neurons are a popular model for investigating mechanisms of neuronal morphogenesis. Da neurons develop in ...

Focal Macropatch Recordings of Synaptic Currents from the Drosophila Larval Neuromuscular Junction

Focal Macropatch Recordings of Synaptic Currents from the Drosophila Larval Neuromuscular Junction

从果蝇幼虫神经肌肉交界处的突触电流的焦 Macropatch 记录

Drosophila neuromuscular junction (NMJ) is an excellent model system to study glutamatergic synaptic transmission. We describe the technique of focal ...