Name: loading drosophila nerve terminals with calcium indicators
Uploaded: 2007-07-30T00:00:00
Description: Watch this Scientific Journal Video about Loading Drosophila Nerve Terminals with Calcium Indicators at JoVE.com

<ol><li>选择一个干净的清扫菜，没有接触过任何固定液。 </li><li>解剖流浪3 路龄果蝇在施耐德的果蝇含有钙的中等幼虫+和L -谷氨酰胺，（不削减任何神经或损伤肌纤维第7，6，13或12） 。 </li><li>选择一个12微米的小费（内径）的玻璃灌装吸管。 </li><li>使用注射器和油管（应用负压吸管）确保不妨碍枪头。 </li><li>选择一个优良的塑料填充丝可插入玻璃吸管?...

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	<li>Macleod, G. T., Hegstrom-Wojtowicz, M., Charlton, M. P., Atwood, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+calcium+signals+in+Drosophila+motor+neuron+terminals.">Fast calcium signals in Drosophila motor neuron terminals.</a> J. Neurophysiol. 88, 2659-2663 (2002).</li><li>Macleod, G. T., Suster, M. L., Charlton, M. P., Atwood, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+neuron+activity+in+the+Drosophila+larval+CNS+detected+with+calcium+indicators.">Single neuron activity in the Drosophila larval CNS detected with calcium indicators.</a> J. Neurosci. Methods. 127, 167-178 (2003).</li><li>Macleod, G. T., Marin, L., Charlton, M. P., Atwood, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+vesicles:+test+for+a+role+in+presynaptic+calcium+regulation.">Synaptic vesicles: test for a role in presynaptic calcium regulation.</a> J. Neurosci. 24, 2496-2505 (2004).</li><li>Reiff, D. F., Thiel, P. R., Schuster, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+regulation+of+active+zone+density+during+long-term+strengthening+of+Drosophila+neuromuscular+junctions.">Differential regulation of active zone density during long-term strengthening of Drosophila neuromuscular junctions.</a> J. Neurosci. 22, 9399-9409 (2002).</li><li>Reiff, D. F., Ihring, A., Guerrero, G., Isacoff, E. Y., Joesch, M., Nakai, J., Borst, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+performance+of+genetically+encoded+indicators+of+neural+activity+in+flies.">In vivo performance of genetically encoded indicators of neural activity in flies.</a> J. Neurosci. 25, 4766-4778 (2005).</li></ol>

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<td>Schneider&#x2019;s Insect Medium</td>
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<td>must contain L-glutamine and calcium</td>
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<td>L-glutamic acid monosodium salt hydrate</td>
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loading drosophila nerve terminals with calcium indicators

钙在神经系统中的许多角色，但没有更令人印象深刻，作为神经递质释放的触发，并没有比支持学习和记忆的突触可塑性的重要使者深刻。为了进一步阐明的分子基础的Ca 2 +依赖的突触机制，需要一个模型系统，基因可塑性和生理访问。果蝇提供了这样一个模型。在这个系统中，基因编码的荧光指标，可用来检测神经末梢的Ca 2 +的变化。然而，这些指标是有限的敏感性钙+和常常表现出非线性响应。合成荧光指标更适合于测量迅速的Ca 2 +与神经活动有关的变化。在这里，我们展示了一个加载到果蝇幼虫生活的神经末梢葡聚糖偶联合成的Ca 2 +指标的技术。特别强调的是放置在协议的那些方面最关键该技术的成功，例如如何以避免沿的隔离神经静电，保持编制的健康在扩展装载期间，，并确保轴突生存提供的Ca 2 +促进密封切断轴突末梢。低亲和力右旋糖酐标记的Ca 2 +的指标，如氟- 4和rhod，这显示了一个高信号的信噪比，同时微创干扰突触前的Ca 2 +动力学。葡聚糖共轭有助于防止钙进入细胞器如线粒体扣押+指标。加载技术可以同样适用于幼虫，胚胎和成人。

Watch this Scientific Journal Video about Loading Drosophila Nerve Terminals with Calcium Indicators at JoVE.com

Loading Drosophila Nerve Terminals with Calcium Indicators

钙是一种在神经系统中无处不在的使者，触发神经递质释放和突触强度的变化。在这里，我们展示了一个加载的Ca 2 +指标到果蝇的神经终端的技术。我们也证明所需设备的制造，并强调技术的成功的关键点。

载入果蝇与神经末梢钙指标

Calcium plays many roles in the nervous system but none more impressive than as the trigger for neurotransmitter release, and none more profound than as ...

Research

JoVE Journal

Biology

Ex vivo Mechanical Loading of Tendon

体外机械装载肌腱

Injuries to the tendon (e.g., wrist tendonitis, epicondyltis) due to overuse are common in sports activities and the workplace.  Most are associated with ...

科研

生物学

Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

可视化的Live果蝇胶质细胞神经​​肌肉交界处，与荧光染料

Our project identified GFP labeled glial structures at the developing larval fly neuromuscular synapse. To look at development of live glial-nerve-muscle ...

Assembly, Loading, and Alignment of an Analytical Ultracentrifuge Sample Cell

大会，加载和分析超速离心机样品池对齐

The analytical ultracentrifuge (AUC) is a powerful biophysical tool that allows us to record macromolecular sedimentation profiles during high speed ...

Dopamine Release at Individual Presynaptic Terminals Visualized with FFNs

多巴胺的释放突触前货箱码头在个别的可视化与FFNs

The nervous system transmits signals between neurons via neurotransmitter release during synaptic vesicle fusion. To observe neurotransmitter uptake and ...

Cargo Loading onto Kinesin Powered Molecular Shuttles

货物装载到驱动蛋白供电分子梭

Cells have evolved sophisticated molecular machinery, such as kinesin motor proteins and microtubule filaments, to support active intracellular transport ...

Whole-Cell Recording of Calcium Release-Activated Calcium (CRAC) Currents in Human T Lymphocytes

Whole-Cell Recording of Calcium Release-Activated Calcium CRAC Currents in Human T Lymphocytes

在人T淋巴细胞的全细胞钙释放激活的钙记录（CRAC）的电流

In T lymphocytes, depletion of Ca2+ from the intracellular Ca2+ store leads to activation of plasmalemmal Ca2+ channels, called Calcium Release-Activated ...

Measuring Fast Calcium Fluxes in Cardiomyocytes

测量心肌细胞的快速钙助焊剂

Cardiomyocytes have multiple Ca2+ fluxes of varying duration that work together to optimize function 1,2. Changes in Ca2+ activity in response to ...

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading TED

直接成像诱导ER钙有针对性的酯酶染料装载（TED）

Visualization of calcium dynamics is important to understand the role of calcium in cell physiology. To examine calcium dynamics, synthetic fluorescent ...

Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+

Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+

监测ER / SR钙释放与目标Ca<sup&gt; 2+</sup&gt;传感器捕捉器<sup&gt; +</sup

Intracellular calcium (Ca2+) transients evoked by extracellular stimuli initiate a multitude of biological processes in living organisms. At the center of ...

Forward Genetic Screen Using Transgenic Calcium Reporter Aequorin to Identify Novel Targets in Calcium Signaling

使用转基因钙处理器 Aequorin 前向基因屏幕，以识别钙信号中的新目标

Forward genetic screens have been important tools in the unbiased identification of genetic components involved in several biological pathways. The basis ...