Name: methods to characterize spontaneous and startle-induced locomotion in a rotenone-induced parkinson's disease model of drosophila
Uploaded: 2014-08-17T14:00:00
Description: Watch this Scientific Journal Video about Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila at JoVE.com

作者要感谢秋里王，语言资源中心，科尔比学院，用于视频处理和埃里克·托马斯，音乐系，科尔比学院技术援助，用于提供背景音乐。该项目由来自国立普通医学科学（P20 GM103423-12）国家研究资源中心，INBRE（P20RR016463-12），资金支持，健康和科学部资助，科尔比学院（STA）的国民研究院。 JL和LWM由来自夏季学者基金，科尔比学院资助。

1， 果蝇惊吓引起的步态分析<ol><li>药物治疗<ol><li>稳重的固定化所需的数字1-3（约8-12）日龄雄蝇使用二氧化碳 ，并将其输送到含有药物的补充食品瓶。注：另麻醉剂如乙醚或冰可用于镇静苍蝇，使数量和处理。 </li><li>允许蝇从镇静在水平位置上恢复20分钟（或直到恢复）与小瓶（防止蝇卡住上的食物），然后竖直放置的管形瓶中在12小时黑暗，12小时光照培养箱中在25℃下...

<ol>
	<li>Olanow, C. W., Tatton, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiology+and+pathogenesis+of+Parkinson's+disease.">Etiology and pathogenesis of Parkinson's disease.</a> Annual review of neuroscience. 22, 123-144 (1999).</li><li>Coulom, H., Birman, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+exposure+to+rotenone+models+sporadic+Parkinson's+disease+in+Drosophila+melanogaster.">Chronic exposure to rotenone models sporadic Parkinson's disease in Drosophila melanogaster.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 24, 10993-10998 (2004).</li><li>Hosamani, R., Ramesh, S. R., Muralidhara, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+rotenone-induced+mitochondrial+oxidative+damage+and+neurotoxicty+in+Drosophila+melanogaster+supplemented+with+creatine.">Attenuation of rotenone-induced mitochondrial oxidative damage and neurotoxicty in Drosophila melanogaster supplemented with creatine.</a> Neurochemical research. 35, 1402-1412 (2010).</li><li>Islam, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+neuroprotective+role+of+the+human+uncoupling+protein+2+(hUCP2)+in+a+Drosophila+Parkinson's+disease+model.">A neuroprotective role of the human uncoupling protein 2 (hUCP2) in a Drosophila Parkinson's disease model.</a> Neurobiology of disease. 46, 137-146 (2012).</li><li>Lawal, H. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Drosophila+vesicular+monoamine+transporter+reduces+pesticide-induced+loss+of+dopaminergic+neurons.">The Drosophila vesicular monoamine transporter reduces pesticide-induced loss of dopaminergic neurons.</a> Neurobiology of. 40, 102-112 (2010).</li><li>St Laurent, ., O'Brien, R., M, L., Ahmad, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sodium+butyrate+improves+locomotor+impairment+and+early+mortality+in+a+rotenone-induced+Drosophila+model+of+Parkinson's+disease.">Sodium butyrate improves locomotor impairment and early mortality in a rotenone-induced Drosophila model of Parkinson's disease.</a> Neuroscience. 246, 382-390 (2013).</li><li>Sherer, T. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+toxicity+in+rotenone+models+of+Parkinson's+disease.">Mechanism of toxicity in rotenone models of Parkinson's disease.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 23, 10756-10764 (2003).</li><li>Munoz-Soriano, V., Paricio, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+models+of+Parkinson's+disease:+discovering+relevant+pathways+and+novel+therapeutic+strategies.">Drosophila models of Parkinson's disease: discovering relevant pathways and novel therapeutic strategies.</a> Parkinson's disease. , 520640 (2011).</li><li>Steffan, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histone+deacetylase+inhibitors+arrest+polyglutamine-dependent+neurodegeneration+in+Drosophila.">Histone deacetylase inhibitors arrest polyglutamine-dependent neurodegeneration in Drosophila.</a> Nature. 413, 739-743 (2001).</li><li>Auluck, P. K., Bonini, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+prevention+of+Parkinson+disease+in+Drosophila.">Pharmacological prevention of Parkinson disease in Drosophila.</a> Nature medicine. 8, 1185-1186 (2002).</li><li>Whitworth, A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+glutathione+S-transferase+activity+rescues+dopaminergic+neuron+loss+in+a+Drosophila+model+of+Parkinson's+disease.">Increased glutathione S-transferase activity rescues dopaminergic neuron loss in a Drosophila model of Parkinson's disease.</a> Proceedings of the National Academy of Sciences of the United States of America. 102, 8024-809 (2005).</li><li>Ahmad, S. T., Steinmetz, S. B., Bussey, H. M., Possidente, B., Seggio, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Larval+ethanol+exposure+alters+free-running+circadian+rhythm+and+per+Locus+transcription+in+adult+D.+melanogaster+period+mutants.">Larval ethanol exposure alters free-running circadian rhythm and per Locus transcription in adult D. melanogaster period mutants.</a> Behavioural brain research. 241, 50-55 (2013).</li><li>Seggio, J. A., Possidente, B., Ahmad, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Larval+ethanol+exposure+alters+adult+circadian+free-running+locomotor+activity+rhythm+in+Drosophila+melanogaster.">Larval ethanol exposure alters adult circadian free-running locomotor activity rhythm in Drosophila melanogaster.</a> Chronobiology international. 29, 75-81 (2012).</li><li>Chaudhuri, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+of+genetic+and+environmental+factors+in+a+Drosophila+parkinsonism+model.">Interaction of genetic and environmental factors in a Drosophila parkinsonism model.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 27, 2457-2467 (2007).</li><li>Feany, M. B., Bender, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+of+Parkinson's+disease.">A Drosophila model of Parkinson's disease.</a> Nature. 404, 394-398 (2000).</li><li>Ali, Y. O., Escala, W., Ruan, K., Zhai, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assaying+locomotor,+learning,+and+memory+deficits+in+Drosophila+models+of+neurodegeneration.">Assaying locomotor, learning, and memory deficits in Drosophila models of neurodegeneration.</a> Journal of Visualized Experiments : JoVE. , (2011).</li><li>Gargano, J. W., Martin, I., Bhandari, P., Grotewiel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+iterative+negative+geotaxis+(RING):+a+new+method+for+assessing+age-related+locomotor+decline+in+Drosophila.">Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila.</a> Experimental gerontology. 40, 386-395 (2005).</li><li>Nichols, C. D., Becnel, J., Pandey, U. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+assay+Drosophila+behavior.">Methods to assay Drosophila behavior.</a> Journal of visualized experiments : JoVE. , (2012).</li><li>Slawson, J. B., Kim, E. Z., Griffith, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+video+tracking+of+locomotion+in+adult+Drosophila+melanogaster.">High-resolution video tracking of locomotion in adult Drosophila melanogaster.</a> Journal of Visualized Experiments : JoVE. , (2009).</li></ol>

在这项研究中，我们描述了两个过程中帕金森病的鱼藤酮诱导的果蝇模型中同时测量长期自发运动和短期惊吓诱发运动。还可以测量在果蝇暴露于已知的建模帕金森氏病如其他药剂，百草枯14，帕金森氏病，例如遗传模型，阿尔法-突触核蛋白突变体15中，并影响运动疾病其他蝇模型这些运动特征。对于这两种方法，替代的方法和修改可以被考虑。?...

运动缺陷是帕金森氏病的主要症状，并在很大程度上引起黑质1的多巴胺能神经元的退化。鱼藤酮是已被广泛研究的模型帕金森运动障碍果蝇 2-6酮类杀虫剂。鱼藤酮通过阻断氧化磷酸化途径，最终导致细胞死亡7导致的氧化损伤。多巴胺能神经元更容易产生鱼藤酮毒性，使得主要马达基于2,7的化学品的影响。通过在果蝇诱发帕金森氏病的症状，我们就可以更好地了解疾病和纠正其症状6,8-11。 果蝇提供了一个很好的模型来研究这种影响，因为它们的基因听话，易于维护，并且具有快速的生命周期。 几项研究已经表明，鱼藤酮引起短期惊跳诱导在果蝇 -当果蝇运动缺陷被保持在鱼藤酮补充食品，他们表现出惊人死不休2-6后以较慢的负geotactic回应。他们未能在小瓶设备，迅速向上攀升对照试验表明了惊吓引起的运动缺陷。 鱼藤酮对长期的效果，自发运动不能很好地说明。 果蝇活动监视器（DAMS）已被成功地用于监测果蝇的昼夜节律运动研究12,13。苍蝇被放置在单独的管，其被加载到DAM。该装置配备有红外线传感器，该计数的次数蝇打破了红外线光束的数目。这些计数可以用作原状的运动和活动12,13的量度。通过将苍蝇在一个坝，鱼藤酮对他们的长期运动的效果可以表征。这项研究描述的方法来MEAS茜短期惊跳诱导运动和长期的自发运动，以便更好地理解的鱼藤酮介导的运动缺陷的影响。对运动缺乏模仿帕金森氏症表征是很重要的，因为它们允许它可以扭转这些运动缺陷等化合物的研究。

<table border="0" cellpadding="0" cellspacing="0" width="628">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Standard narrow vials</td>
			<td style="width:127px;">Genesee Scientific</td>
			<td style="width:119px;">32-120</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Rotenone</td>
			<td style="width:127px;">Sigma</td>
			<td style="width:119px;">R8875</td>
			<td>Store in freezer, make fresh for each experiment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Dimethyl Sulfoxide (DMSO)</td>
			<td style="width:127px;">Sigma</td>
			<td style="width:119px;">D8418</td>
			<td>Solvent for rotenone</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Instant Drosophila medium</td>
			<td style="width:127px;">Carolina Biological&#160;</td>
			<td style="width:119px;">Formula 4-24</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Drosophila activity monitor (DAM)</td>
			<td style="width:127px;">Trikinetics</td>
			<td style="width:119px;">DAM2</td>
			<td>trikinetics.com</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">DAM tubes</td>
			<td style="width:127px;">Trikinetics</td>
			<td style="width:119px;">Tubes 5X65 mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;"></td>
			<td style="width:127px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Recipe for Rotenone +food (125 mM dose)</td>
			<td style="width:127px;"></td>
			<td style="width:119px;"></td>
			<td>Make 62.5 mM rotenone stock solution in DMSO by dissolving 25 mg rotenone in 1 ml DMSO&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:127px;"></td>
			<td style="width:119px;"></td>
			<td>For 125 mM dose, add 10 mM rotenone stock in DMSO to 5 ml water.</td>
		</tr>
	</tbody>
</table>

methods to characterize spontaneous and startle-induced locomotion in a rotenone-induced parkinson's disease model of drosophila

帕金森病是一种神经退行性疾病而导致的多巴胺能神经元的变性，在中枢神经系统中，主要在黑质。这种疾病会导致电机的不足，这表现为强直，震颤和老年痴呆症的人。鱼藤酮是一种杀虫剂，通过抑制线粒体的电子传递链的功能引起的氧化损伤。它也被用来建模帕金森氏病的果蝇 。苍蝇有一个固有的负面geotactic反应，这迫使他们在被吓了一跳向上攀升。它已经确定，鱼藤酮引起早期死亡率和运动缺陷扰乱果蝇的爬它们已被窃听向下之后能力。然而，鱼藤酮对自发运动的效果没有记载。本研究概述了两个敏感，重复性好，高通量检测的表征鱼藤酮诱导的缺陷短期惊跳诱导运动和长期自发运动的果蝇 。这些测定法可便利地适用于表征运动缺陷和治疗剂的效力的其他果蝇模型。

果蝇惊吓引起的步态分析 野生型， 州-S，苍蝇30秒（ 图1）后表现出的双管形瓶装置的仅约88％和苍蝇的顶部和底部部分的5％的鲁棒负geotactic响应分别。暴露于125μM和250μM的鱼藤酮3天蝇表明在果蝇中的顶端部分，并在果蝇中的底部的数目略有增加的数量略有减少。暴露于500μM的鱼藤酮蝇显示显著缺陷在负geotactic反应（P &lt;0.05 ANOVA，邦弗...

Watch this Scientific Journal Video about Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila at JoVE.com

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila

方法来描述自发和惊吓引起的步态中的鱼藤酮诱导的帕金森病模型<em&gt;果蝇</em

帕金森病是一种神经退行性疾病而导致的多巴胺能神经元在中枢神经系统的变性，造成运动缺陷。鱼藤酮车型帕金森氏症的果蝇 。本文概述了两种检测的特点所造成的鱼藤酮是自发性的，惊吓引起的运动缺陷。

Parkinson&#8217;s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, ...

Research

JoVE Journal

Neuroscience

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

自体血液的注入模型小鼠自发性脑出血

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model.  While ICH
accounts for 10-15% ...

科研

神经科学

Methods to Assay Drosophila Behavior

Methods to Assay Drosophila Behavior

对检测方法果蝇行为

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

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

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

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 ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

在自发活动的测定<em&gt;果蝇</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

In Vivo Single-Molecule Tracking at the Drosophila Presynaptic Motor Nerve Terminal

In Vivo Single-Molecule Tracking at the Drosophila Presynaptic Motor Nerve Terminal

在体内果蝇前运动神经末端的单分子跟踪

An increasing number of super-resolution microscopy techniques are helping to uncover the mechanisms that govern the nanoscale cellular world. ...

Quantifying Spontaneous Ca2+ Fluxes and their Downstream Effects in Primary Mouse Midbrain Neurons

Quantifying Spontaneous Ca2+ Fluxes and their Downstream Effects in Primary Mouse Midbrain Neurons

定量自发 Ca2+ 通量及其在原发性小鼠中脑神经元中的下游效应

Parkinson&#8217;s disease (PD) is a devastating neurodegenerative disorder caused by the degeneration of dopaminergic (DA) neurons. Excessive Ca2+ influx ...

Applying the RatWalker System for Gait Analysis in a Genetic Rat Model of Parkinson's Disease

将RatWalker系统应用于帕金森病遗传大鼠模型中的步态分析

Parkinson's disease (PD) is a progressive neurodegenerative disorder caused by the loss of dopaminergic (DA) neurons in the substantia nigra pars ...

The 6-hydroxydopamine Rat Model of Parkinson's Disease

帕金森病的6-羟基多巴胺大鼠模型

Motor symptoms of Parkinson's disease (PD)-bradykinesia, akinesia, and tremor at rest-are consequences of the neurodegeneration of dopaminergic neurons in ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

左旋多巴诱导的帕金森病大鼠模型中运动障碍的诱导与评估

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

Analyzing the Parkinson's Disease Mouse Model Induced by Adeno-associated Viral Vectors Encoding Human &#945;-Synuclein

Analyzing the Parkinson's Disease Mouse Model Induced by Adeno-associated Viral Vectors Encoding Human ÃƒÅ½Ã‚Â±-Synuclein

分析编码人类α突触核蛋白的腺相关病毒载体诱导的帕金森病小鼠模型

Parkinson&#39;s disease is a neurodegenerative disorder that involves the death of the dopaminergic neurons of the nigrostriatal pathway and, ...