Name: localization, identification, and excision of murine adipose depots
Uploaded: 2014-12-04T14:00:00
Description: Watch this Scientific Journal Video about Localization, Identification, and Excision of Murine Adipose Depots at JoVE.com

The authors have no acknowledgements.

注：所有动物的程序都与辛辛那提大学的机构动物护理和使用委员会（IACUC），并按照该指南实验动物的护理和使用来自美国国立卫生研究院（NIH出版号的审批进行，85-23，1996年修订）。 1.安乐死和消毒鼠标<ol><li>鼠标放置在含有异氟醚的supratheraputic剂量的Dropbox，并允许吸气效果。一旦老鼠被安乐死，从包装盒中取出。 </li><li>与颈部脱臼安乐死的第二个手段。 </li><li>由清?...

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	<li>Ogden, C. L., Carroll, M. D., Kit, B. K., Flegal, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+of+obesity+in+the+United+States,+2009-2010.">Prevalence of obesity in the United States, 2009-2010.</a> NCHS Data Brief. 82, 1-8 (2012).</li><li>Flegal, K. M., Carroll, M. D., Kit, B. K., Ogden, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+of+obesity+and+trends+in+the+distribution+of+body+mass+index+among+US+adults,+1999-2010.">Prevalence of obesity and trends in the distribution of body mass index among US adults, 1999-2010.</a> 307 (5), 491-497 (2012).</li><li>Cawley, J., Meyerhoefer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+medical+care+costs+of+obesity:+an+instrumental+variables+approach.">The medical care costs of obesity: an instrumental variables approach.</a> Journal of Health Economics. 31 (1), 219-230 (2012).</li><li>Marder, W., Chang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Childhood+Obesity:+Costs,+Treatment+Patterns,+Disparities+in+Care,+and+Prevalent+Medical+Conditions.">Childhood Obesity: Costs, Treatment Patterns, Disparities in Care, and Prevalent Medical Conditions.</a> Thomson Medstat Research Brief. , (2006).</li><li>Cortez, M., Carmo, L. S., Rogero, M. M., Borelli, P., Fock, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-fat+diet+increases+IL-1,+IL-6,+and+TNF-&#945;+production+by+increasing+NF-&#954;B+and+attenuating+PPAR-&#947;+expression+in+bone+marrow+mesenchymal+stem+cells.">A high-fat diet increases IL-1, IL-6, and TNF-&#945; production by increasing NF-&#954;B and attenuating PPAR-&#947; expression in bone marrow mesenchymal stem cells.</a> Inflammation. 36 (2), 379-386 (2013).</li><li>Sanchez-Gurmaches, J., Hung, C. M., Sparks, C. A., Tang, Y., Li, H., Guertin, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PTEN+loss+in+the+Myf5+lineage+redistributes+body+fat+and+reveals+subsets+of+white+adipocytes+that+arise+from+Myf5+precursors.">PTEN loss in the Myf5 lineage redistributes body fat and reveals subsets of white adipocytes that arise from Myf5 precursors.</a> Cell Metab. 16 (3), 348-362 (2012).</li><li>Sethi, J. K., Vidal-Puig, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thematic+review+series:+adipocyte+biology.+Adipose+tissue+function+and+plasticity+orchestrate+nutritional+adaptation.">Thematic review series: adipocyte biology. Adipose tissue function and plasticity orchestrate nutritional adaptation.</a> J Lipid Res. 48 (6), 1253-1262 (2007).</li><li>Aarsland, A., Chinkes, D., Wolfe, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+and+whole-body+fat+synthesis+in+humans+during+carbohydrate+overfeeding.">Hepatic and whole-body fat synthesis in humans during carbohydrate overfeeding.</a> Am J Clin Nutr. 65 (6), 1774-1782 (1997).</li><li>Park, A., Kim, W. K., Bae, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinction+of+white,+beige+and+brown+adipocytes+derived+from+mesenchymal+stem+cells.">Distinction of white, beige and brown adipocytes derived from mesenchymal stem cells.</a> World J. Stem Cells. 6 (1), 33-42 (2014).</li><li>Wu, 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=Beige+adipocytes+are+a+distinct+type+of+thermogenic+fat+cell+in+mouse+and.">Beige adipocytes are a distinct type of thermogenic fat cell in mouse and.</a> 150 (2), 366-376 (2012).</li><li>Giralt, M., Villarroya, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=White,+brown,+beige/brite:+different+adipose+cells+for+different+functions.">White, brown, beige/brite: different adipose cells for different functions.</a> Endocrinology. 154 (9), 2992-3000 (2013).</li><li>Casteilla, L., P&#233;nicaud, L., Cousin, B., Calise, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choosing+an+adipose+tissue+depot+for+sampling:+factors+in+selection+and+depot+specificity.">Choosing an adipose tissue depot for sampling: factors in selection and depot specificity.</a> Methods Mol. Biol. 456, 23-38 (2008).</li><li>Grant, R., Youm, Y. H., Ravussin, A., Dixit, V. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+adipose+tissue+leukocytosis+in+obesity.">Quantification of adipose tissue leukocytosis in obesity.</a> Methods Mol. Biol. 1040, 195-209 (2013).</li></ol>

肥胖可能导致大的宿主疾病发病率和该脂肪起着尚不完全清楚的作用的充分理解;因此继续研究在脂肪的字段是必要的。动物模型，特别是小鼠模型非常适合在疾病和潜在的药物治疗试验的进展最初的研究。在使用这些模型，精确的隔离和脂肪车厂切除脂肪影响了疾病的病理研究中的一个非常重要和必要的工具。 在关于脂肪库目前的文献，有文献关于代谢行为和脂肪仓库之间?...

脂肪做了一个显着的出现在媒体的聚光灯下，由于在过去几十年的20世纪肥胖的急剧增加。目前，肥胖的影响超过三分之一的成年人和在美国（美国）1儿童和青少年的17％。跨越各族，周围肥胖的统计研究表明，非西班牙裔黑人有肥胖（49.5％）的最高年龄调整率与墨西哥裔美国人（40.4％），所有西班牙裔（39.1％），和非比较西班牙裔白人（34.3％），2。肥胖的经济效应也为医疗保健系统日益受到关注。在2012年，据估计，每年的医疗照顾，肥胖在美国，2005年费用为190.2十亿，近21％的整体医疗支出预算。可悲的是，儿童肥胖，估计是负责单独$ 14十亿的直接医疗费用。在统计上，它被确定的平均医疗费用肥胖人士是2741美元高于去年比没有这种发病率3-5。 肥胖是用于各种诸如条件的主要危险因素：2型糖尿病，血脂异常，心血管疾病，癌症，肌肉骨骼疾病和慢性炎症。肥胖是深深依赖于代谢综合征等慢性疾病6-8的发病机制。随着肥胖等合并症之间的这种激烈的负极连接，科研集中注意力，以便更好地了解当前的流行和脂肪所扮演的多样和重要的角色。 从历史上看，脂肪组织被认为是无关紧要的，并仅仅看作是一个简单的填充组织。目前，脂肪已被证明在体内的功能中发挥许多重要作用：代谢，激素调节，炎症，保护和绝缘9。脂肪组织主要由脂肪细胞还含有周细胞，血管内皮细胞，单核细胞，巨噬细胞和多能干细胞8。脂肪组织是分布在整个身体中不同的仓库。主体仓库可以皮下发现，皮下，肌肉内，和本能10。脂肪库已被证明有仓库特定的代谢概况，从而衍生出一个仓库特定易感性肥胖及相关病症8。 传统上，脂肪组织已被分为两种主要类型：白色脂肪组织（WAT）和棕色脂肪组织（BAT）;虽然最近的文献表明，第三组的派驻命名BRITE或米色脂肪11。脂肪组织已被证明具有不同的颜色，形态，代谢功能，生化特征和表达10的遗传模式。在WAT脂肪细胞有一个单一的，大的脂滴和线粒体的数量可变。 WAT在身体的皮下和内脏地方是显性找到。 WAT功能主要是作为能量储存和器官保护的部位。在BAT脂肪细胞有一个多房形态和丰富的线粒体。 BAT位于主要在颈部和胸部的大血管，以及肩胛12。 BAT主要功能是调节产热7能源花费的行为。 BRITE或米色脂肪已经显示的，共享一个类似的形态和表达BAT但已被发现是源自白色脂肪细胞11。 在这个手稿中描述的手术方法为研究人员提供的能力，分析不同效果的因素，如：环境，医药，和遗传学，对脂肪;以及有益或有害作用的脂肪起着病和总体健康状况。此外，提供了一种方法来鉴定和分离不同类型的脂肪组织，以以便更好地了解车厂之间的生化关系和差异。这可以在确定身体内的位置，功能，和类型的脂肪之间的关系提供帮助。该所述方法通过提供用于毛可视化，基因表达分析，蛋白质表达分析，组织学检查，以及初级细胞系的隔离体外研究实现这一目的。目前，有许多文章，提供深入了解不同脂肪库，以及它们的解剖位置的代谢行为;但不提供一个深入的方法，对如何具体定位，识别和隔离这些仓库。这种手术方法提供了精确的技术，其允许多个站的具有夹层和污染的最小量相比较，以设计用于一个或两个仓库13-14的分离方法更为隔离。 该协议的目的是提供精确的方法对不同类型的多个解剖位置脂肪库的鉴定和分离。

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			<td height="21" style="height:21px;">isoflurane</td>
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localization, identification, and excision of murine adipose depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

识别和腹股沟皮下脂肪，肩胛间褐色脂肪，内脏附睾脂肪（ 图1），以及主动脉弓血管周围脂肪，胸主动脉脂肪，肾上主动脉脂肪和下腹主动脉脂肪的本地化（ 图2），使用取得了成功所描述的手术方法。 BAT和WAT样品之间的组织学检查和分化采用HE染色（H＆E）染色（ 图3）进行了积极评价。脂联素（ADIPOQ），过氧化物酶体增殖物激活受体γ（PPAR-γ），的RNA水?...

Watch this Scientific Journal Video about Localization, Identification, and Excision of Murine Adipose Depots at JoVE.com

Localization, Identification, and Excision of Murine Adipose Depots

由于肥胖等合并症之间的激烈负极连接，对脂肪的作用发挥的研究在疾病和整体健康是必要的。我们提出了一个协议，用于分离和脂肪仓库允许利用原位脂肪研究的切除和体外方法。

本地化识别，切除脂肪小鼠油库，并

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

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