Name: insulin injection and hemolymph extraction to measure insulin sensitivity in adult drosophila melanogaster
Uploaded: 2011-06-30T13:00:00
Description: Watch this Scientific Journal Video about Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster at JoVE.com

这项工作是从NIA的YW.CF（AG21068，AG31086）的赠款支持。

1。胰岛素溶液的制备<ol><li>准备新鲜牛胰岛素溶液溶解在PBS中的胰岛素，达到0.01毫克/毫升的浓度。胰岛素/ PBS和控制PBS解决方案必须保持整个注射过程中的冰。这些解决方案应与0.5％（V / V）FD＆C蓝色没有准备。 1食品着色剂。 </li></ol> 2。针的制备和注射设置<ol><li>使用一个车夫设置产生足够的质量针注射微量puller.The：热火，345;拉，210;速度，100;时间，200（100毫秒），准备?...

<ol>
	<li>Rulifson, E. J., Kim, S. K., Nusse, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ablation+of+insulin-producing+neurons+in+flies:+growth+and+diabetic+phenotypes.">Ablation of insulin-producing neurons in flies: growth and diabetic phenotypes.</a> Science. 296, 1118-1118 (2002).</li><li>Kim, S. K., Rulifson, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conserved+mechanisms+of+glucose+sensing+and+regulation+by+Drosophila+corpora+cardiaca+cells.">Conserved mechanisms of glucose sensing and regulation by Drosophila corpora cardiaca cells.</a> Nature. 431, 316-316 (2004).</li><li>Broughton, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longer+lifespan,+altered+metabolism,+and+stress+resistance+in+Drosophila+from+ablation+of+cells+making+insulin-like+ligands.">Longer lifespan, altered metabolism, and stress resistance in Drosophila from ablation of cells making insulin-like ligands.</a> Proc Natl Acad Sci U S A. 102, 3105-3105 (2005).</li><li>Haselton, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+ablation+of+adult+Drosophila+insulin-producing+neurons+modulates+glucose+homeostasis+and+extends+life+span+without+insulin+resistance.">Partial ablation of adult Drosophila insulin-producing neurons modulates glucose homeostasis and extends life span without insulin resistance.</a> Cell Cycle. 9, 3063-3063 (2010).</li><li>Haselton, A. T., Fridell, Y. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+Drosophila+melanogaster+as+a+model+for+the+study+of+glucose+homeostasis.">Adult Drosophila melanogaster as a model for the study of glucose homeostasis.</a> Aging (Albany NY). 2, 523-523 (2010).</li><li>Belgacem, Y. H., Martin, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+insulin+pathways+alters+trehalose+level+and+abolishes+sexual+dimorphism+in+locomotor+activity+in+Drosophila.">Disruption of insulin pathways alters trehalose level and abolishes sexual dimorphism in locomotor activity in Drosophila.</a> J Neurobiol. 66, 19-19 (2006).</li><li>Demerec, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biology of Drosophila. , (1950).</li></ol>

在本报告中描述的技术在任何调查在果蝇血淋巴成分的检测的改变导致生理过程的研究，是潜在有用的。通过以这种方式相结合的注射和血淋巴收集，它是可能的，以确定一个特定的实验性治疗或操纵的切身有关生理影响。这种“放血”技术的主要优点是收集超过以前的技 ​​术，涉及斩首 2血淋巴，这种技术最大限度地减少污染血淋巴样品与肠道contents.If护理是不收取任何pericereb...

<table border="1"><tbody><tr><th>试剂/设备名称</th><th>公司</th><th>目录编号</th></tr><tr><td>牛胰岛素</td><td>西格玛</td><td> I5500 </td></tr><tr><td>无限葡萄糖试剂</td><td>热电公司</td><td> TR1541 </td></tr><tr><td>手动微量</td><td>萨特仪器</td><td></td></tr><tr><td> P - 87 Flamming /布朗微量拉马</td><td>萨特仪器</td><td></td></tr><tr><td>单桶毛细管硼硅玻璃</td><td>上午系统</td><td> 626000 </td></tr><tr><td> FD＆C蓝色1号</td><td>的McCormick＆公司</td><td></td></tr><tr><td> 1μLmicrocapillary管</td><td>德拉蒙德</td><td></td></tr><tr><td>三轴手动显微操纵器和基</td><td>世界精密仪器</td><td></td></tr></tbody></table>

insulin injection and hemolymph extraction to measure insulin sensitivity in adult drosophila melanogaster

保守的营养感应机制之间存在着哺乳动物和果蝇肽类似哺乳动物的胰岛素和胰高血糖素，分别在幼虫阶段发育1,2维持血糖平衡的功能。主要有丝分裂后的成年果蝇的研究发现扰动糖代谢的胰岛素生产细胞（IPCS）3样肽（ILP）的遗传消融的结果。因此，成年果蝇保持一个合适的代谢紊乱，包括II型糖尿病的遗传模型系统的巨大潜力。为了进一步发展果蝇系统，用于测量哺乳动物糖耐量和胰岛素敏感性的可比生理检测，必须建立。为此，我们最近测量口服葡萄糖容忍的反应在成人飞介绍一种新的程序，并表现出成人的IPC的重要性，在维持血糖稳态4,5。在这里，我们已经修改为先前所描述的胰岛素注射6的程序，并结合血淋巴中提取了一种新的方法来测量在成人飞周围胰岛素敏感性。与众不同的是，我们的协议允许成年果蝇的能力，处置周边葡萄糖负荷时注射胰岛素的方法，使胰岛素信号突变和影响成人飞糖耐量和胰岛素敏感性的潜在干预是可行的特点直接的生理测量。

Watch this Scientific Journal Video about Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster at JoVE.com

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

保守的胰岛素信号通路在果蝇中发现<em&gt;果蝇</em&gt;使这个有机体建模，包括II型糖尿病的代谢紊乱的潜在工具。为此，关键是建立生理检测在成人飞周边葡萄糖处置全身胰岛素的作用，以有效衡量。

注射胰岛素和血淋巴的提取来衡量成人胰岛素敏感性果蝇</em

Conserved nutrient sensing mechanisms exist between mammal and fruit fly where peptides resembling mammalian insulin and glucagon, respectively function ...

Research

JoVE Journal

Biology

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

高分辨率视频跟踪移动的，在成年果蝇。

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

科研

生物学

Dissection of Oenocytes from Adult Drosophila melanogaster

Dissection of Oenocytes from Adult Drosophila melanogaster

从成人Oenocytes剖析果蝇</em

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides ...

A Quantitative Assay for Insulin-expressing Colony-forming Progenitors

一个定量测定胰岛素表达细胞集落形成先觉

The field of pancreatic stem and progenitor cell biology has been hampered by a lack of in vitro functional and quantitative assays that allow for the ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

寿命测量<em&gt;果蝇</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Use of the Ramsay Assay to Measure Fluid Secretion and Ion Flux Rates in the Drosophila melanogaster Malpighian Tubule

Use of the Ramsay Assay to Measure Fluid Secretion and Ion Flux Rates in the Drosophila melanogaster Malpighian Tubule

使用拉姆齐试验来测量液体分泌和离子流率在<em&gt;果蝇</em&gt;马氏管

Modulation of renal epithelial ion transport allows organisms to maintain ionic and osmotic homeostasis in the face of varying external conditions. The ...

Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes

Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes

葡萄糖摄取测量和对胰岛素刺激的反应<em&gt;体外</em&gt;培养的人类初级肌管

Skeletal muscle is the largest glucose deposit in mammals and largely contributes to glucose homeostasis. Assessment of insulin sensitivity of muscle ...

Confocal Imaging of Neuropeptide Y-pHluorin: A Technique to Visualize Insulin Granule Exocytosis in Intact Murine and Human Islets

神经肽 Y-pHluorin 的共焦成像: 一种完整的小鼠和人胰岛胰岛素颗粒胞的可视化技术

Insulin secretion plays a central role in glucose homeostasis under normal physiological conditions as well as in disease. Current approaches to study ...

Optical Cross-Sectional Muscle Area Determination of Drosophila Melanogaster Adult Indirect Flight Muscles

Optical Cross-Sectional Muscle Area Determination of Drosophila Melanogaster Adult Indirect Flight Muscles

果蝇成人间接飞行肌的光学横截肌面积测定

Muscle mass wasting, known as muscle atrophy, is a common phenotype in Drosophila models of neuromuscular diseases. We have used the indirect flight ...

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

可视化和跟踪活果蝇黑色素细胞卵室的内源性 mRNA

Fluorescence-based imaging techniques, in combination with developments in light microscopy, have revolutionized how cell biologists conduct live cell ...

Measuring Relative Insulin Secretion using a Co-Secreted Luciferase Surrogate

使用共同分泌的Luciferase代理物测量相对胰岛素分泌

Performing antibody-based assays for secreted insulin post-sample collection usually requires a few hours to a day of assay time and can be expensive, ...