Name: body composition and metabolic caging analysis in high fat fed mice
Uploaded: 2018-05-24T14:00:00
Description: Watch this Scientific Journal Video about Body Composition and Metabolic Caging Analysis in High Fat Fed Mice at JoVE.com

我们感谢阿尔弗雷德医学研究和教育局动物服务小组 (AMREP) 的工作人员对这项研究中使用的小鼠的援助和照顾, 并支持维多利亚国家的运营基础设施支持计划。政府。

所描述的所有实验都得到了阿尔弗雷德医学研究教育局动物伦理委员会 (AMREP) 的批准, 并根据澳大利亚国家卫生和医学研究理事会 (澳洲) 提供了人道护理, 以指导动物实验。动物被管理他们的规定的饮食和水ad 随意和安置在温度控制的环境 (~ 21-22 °c) 以 12 h 光和 12 h 黑暗的周期。七周大的雄性小鼠 (在 C57Bl/6J 背景下) 喂食正常的饮食 (能量含量 14.3 MJ/千克, 包括76% 焦从碳水化合物, 5% 脂肪, 19%...

<ol>
	<li>Chen, W., Wilson, J. L., Khaksari, M., Cowley, M. A., Enriori, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abdominal+fat+analyzed+by+DEXA+scan+reflects+visceral+body+fat+and+improves+the+phenotype+description+and+the+assessment+of+metabolic+risk+in+mice.">Abdominal fat analyzed by DEXA scan reflects visceral body fat and improves the phenotype description and the assessment of metabolic risk in mice.</a> Am J Physiol Endocrinol Metab. 303 (5), E635-E643 (2012).</li><li>Kovner, I., Taicher, G. Z., Mitchell, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calibration+and+validation+of+EchoMRI+whole+body+composition+analysis+based+on+chemical+analysis+of+piglets,+in+comparison+with+the+same+for+DXA.">Calibration and validation of EchoMRI whole body composition analysis based on chemical analysis of piglets, in comparison with the same for DXA.</a> Int J Body Compos Res. 8 (1), 17-29 (2010).</li><li>EchoMRI. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Software User Manual: Whole body composition analyzer. , (2016).</li><li>Columbus Instruments. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Oxymax for Windows User Manual. , (2014).</li><li>Tschop, M. H., 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+guide+to+analysis+of+mouse+energy+metabolism.">A guide to analysis of mouse energy metabolism.</a> Nat Methods. 9 (1), 57-63 (2011).</li><li>Speakman, 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=Measuring+energy+metabolism+in+the+mouse+-+theoretical,+practical,+and+analytical+considerations.">Measuring energy metabolism in the mouse - theoretical, practical, and analytical considerations.</a> Front Physiol. 4, (2013).</li><li>Swoap, S. 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=Vagal+tone+dominates+autonomic+control+of+mouse+heart+rate+at+thermoneutrality.">Vagal tone dominates autonomic control of mouse heart rate at thermoneutrality.</a> Am J Physiol Heart Circ Physiol. 294 (4), H1581-H1588 (2008).</li><li>Tian, X. Y., 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=Thermoneutral+housing+accelerates+metabolic+inflammation+to+potentiate+atherosclerosis+but+not+insulin+resistance.">Thermoneutral housing accelerates metabolic inflammation to potentiate atherosclerosis but not insulin resistance.</a> Cell Metab. 23 (1), 165-178 (2016).</li><li>Giles, D. 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=Thermoneutral+housing+exacerbates+nonalcoholic+fatty+liver+disease+in+mice+and+allows+for+sex-independent+disease+modeling.">Thermoneutral housing exacerbates nonalcoholic fatty liver disease in mice and allows for sex-independent disease modeling.</a> Nat Med. 23 (7), 829-838 (2017).</li><li>Lee, M. W., 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=Activated+type+2+innate+lymphoid+cells+regulate+beige+fat+biogenesis.">Activated type 2 innate lymphoid cells regulate beige fat biogenesis.</a> Cell. 160 (1-2), 74-87 (2015).</li><li>Kusminski, C. M., 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=MitoNEET-driven+alterations+in+adipocyte+mitochondrial+activity+reveal+a+crucial+adaptive+process+that+preserves+insulin+sensitivity+in+obesity.">MitoNEET-driven alterations in adipocyte mitochondrial activity reveal a crucial adaptive process that preserves insulin sensitivity in obesity.</a> Nat Med. 18 (10), 1539-1549 (2012).</li><li>Judex, 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=Quantification+of+adiposity+in+small+rodents+using+micro-CT.">Quantification of adiposity in small rodents using micro-CT.</a> Methods. 50 (1), 14-19 (2010).</li><li>Chaurasia, 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=Adipocyte+ceramides+regulate+subcutaneous+adipose+browning,+inflammation,+and+metabolism.">Adipocyte ceramides regulate subcutaneous adipose browning, inflammation, and metabolism.</a> Cell Metab. 24 (6), 820-834 (2016).</li><li>Matthews, V. 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=Interleukin-6-deficient+mice+develop+hepatic+inflammation+and+systemic+insulin+resistance.">Interleukin-6-deficient mice develop hepatic inflammation and systemic insulin resistance.</a> Diabetologia. 53 (11), 2431-2441 (2010).</li><li>Tschop, M., Smiley, D. L., Heiman, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ghrelin+induces+adiposity+in+rodents.">Ghrelin induces adiposity in rodents.</a> Nature. 407 (6806), 908-913 (2000).</li><li>Garcia, M. C., 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=Mature-onset+obesity+in+interleukin-1+receptor+I+knockout+mice.">Mature-onset obesity in interleukin-1 receptor I knockout mice.</a> Diabetes. 55 (5), 1205-1213 (2006).</li><li>Kowalski, G. M., Bruce, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+regulation+of+glucose+metabolism:+Implications+and+considerations+for+the+assessment+of+glucose+homeostasis+in+rodents.">The regulation of glucose metabolism: Implications and considerations for the assessment of glucose homeostasis in rodents.</a> Am J Physiol Endocrinol Metab. 307 (10), E859-E871 (2014).</li><li>McGuinness, O. P., Ayala, J. E., Laughlin, M. R., Wasserman, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIH+experiment+in+centralized+mouse+phenotyping:+the+Vanderbilt+experience+and+recommendations+for+evaluating+glucose+homeostasis+in+the+mouse.">NIH experiment in centralized mouse phenotyping: the Vanderbilt experience and recommendations for evaluating glucose homeostasis in the mouse.</a> Am J Physiol Endocrinol Metab. 297 (4), E849-E855 (2009).</li></ol>

关键步骤
本文所述的协议提供了一个例子, 其中的方法, 以测量身体成分和各种代谢参数的小鼠使用身体成分分析仪和代谢动物监测系统。对于这两种技术, 至关重要的是要确保机器的工作是最佳的, 而且要做到这一点, 研究人员必须对人体成分分析仪进行系统测试, 并校准到已知的代谢气体成分。动物监测系统使用前的设备。这将确保结果更加一致, 并有机会发现?...

新陈代谢支撑着正常细胞、器官和全身生理学的许多方面。因此, 在各种病理的设置中, 新陈代谢的改变可能直接导致潜在的状况, 或者可能会受到负面影响, 作为病理的副作用。传统上, 代谢研究和能量平衡研究集中在肥胖和相关的疾病领域, 如胰岛素抵抗、糖尿病前、葡萄糖不耐受、心血管疾病和糖尿病。鉴于世界范围内这种情况不断升级, 以及这些条件所造成的个人、社会和经济代价, 这项研究是有必要的。因此, 制定预防战略和针对肥胖的新疗法是世界各地研究实验室的一个持续目标, 这些研究主要依赖于临床前老鼠模型。
虽然称重小鼠对体重的增加或损失提供了可靠的评估, 但它并没有提供构成全身成分 (脂肪质量、精益质量、游离水以及毛皮和爪子等其他成分) 的不同成分的分解。在研究完成后, 脂肪垫的重量在老鼠死后提供了一个准确的测量不同的脂肪库, 但只能提供一个单一的时间点的数据。因此, 经常需要招收多个群体, 以调查随着时间的推移, 肥胖的发展, 大大增加动物数量, 时间和成本。使用双能量 X 射线骨密度测定 (DEXA) 提供了一种方法来评估身体脂肪和瘦组织的内容, 并允许研究员获得数据的纵向方式。然而, 这一过程要求老鼠麻醉1, 反复的麻醉可能会影响脂肪组织的积聚或影响新陈代谢调节的其他方面。EchoMRI 利用核磁共振 relaxometry 测量脂肪和精益质量、游离水和总含水量。这是可以实现的, 因为不同的组织成分之间的对比, 在时间, 振幅和空间分布的产生的无线电频率允许划定和定量的每一种组织类型的差异。这种技术是有利的, 因为它是非侵入性的, 快速的, 简单的, 不需要麻醉或辐射, 而且, 重要的是, 已积极验证抗化学分析2。
肥胖的一个关键考虑和相关的研究是能量平衡等式。而脂肪堆积比纯粹的能量更复杂 (食物摄入) 与能量 (能量消耗) 相比, 它们是能够测量的重要因素。每日能量开支是一共四个不同的组分: (1) 基本的能量开支 (休息的新陈代谢率);(2) 由于食品消耗的热效应, 能源支出;(3) 体温所需的能量;(4) 体力活动所消耗的能量。当能源消耗产生热量时, 用一种动物 (称为直接热量计) 来测量热量, 可以用来评估能源支出。或者, 对 o2和 co2的受启发和过期浓度的测量, 允许确定全体 O2消耗量和 co2生产, 可作为间接测量的一种方法 (间接热计量) 加热生产, 从而计算能源消耗。食物摄入量的增加或能源支出的减少将使老鼠容易发胖, 观察这些参数的变化可以为特定的肥胖模型提供可能的行动机制的有用信息。一个相关的代谢参数的兴趣是呼吸交换比 (汇率), 一个指标的基质/燃料 (即,碳水化合物或脂肪) 的比例正在进行新陈代谢和被用来产生能量。因此, 测量食物摄入量 (能量消耗) 与体力活动水平相结合, O2消费、汇率和能源支出可以提供对机体代谢剖面的广泛理解。收集这些数据的一种方法是使用一个综合的实验动物监测系统 (蛤), 它是基于间接量热法测量能量消耗, 并具有确定物理活动水平的额外能力 (光束断裂) 和食物摄取通过秤纳入测量室。
在本议定书中, 我们提供了一个直接的描述, 使用人体成分分析仪来评估小鼠体内成分和代谢动物监测系统, 以衡量新陈代谢的各个方面。将讨论这些技术的考虑和限制, 以及建议的分析、解释和数据表示方法。

<table border="0" cellpadding="0" cellspacing="0" width="1053">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:251px;">4 in 1 system</td>
			<td style="width:157px;">EchoMRI</td>
			<td style="width:275px;">4 in 1 system</td>
			<td style="width:371px;">Whole body composition analyser</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Canola oil test sample (COSTS)</td>
			<td>EchoMRI</td>
			<td style="width:275px;">Mouse-specific (contact company for cat number)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Animal specimen holder&#160;</td>
			<td>EchoMRI</td>
			<td style="width:275px;">103-E56100R</td>
			<td style="width:371px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:251px;">Delimiter&#160;</td>
			<td>EchoMRI</td>
			<td style="width:275px;">600-E56100D</td>
			<td style="width:371px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">12 chamber system</td>
			<td>Columbus Instruments</td>
			<td>Custom built</td>
			<td>Metabolic Caging System; includes control program</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Drierite</td>
			<td>Fisher Scientific</td>
			<td>238988</td>
			<td>CLAMS consumable</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Calibration gas tank</td>
			<td>Air Liquide</td>
			<td>Mixed to order</td>
			<td>Gas calibration (0.5% CO2, 20.5% O2, balance nitrogen).&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal chow diet</td>
			<td>Specialty Feeds</td>
			<td>Irradiated mouse and rat diet</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">High fat diet</td>
			<td>Specialty Feeds</td>
			<td>SF04-001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Balance</td>
			<td>Mettler Toledo</td>
			<td>PL202-S</td>
			<td>Balance for weighing mice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TexQ Disinfectant spray</td>
			<td>TexWipe</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hydrogen Peroxide cleaning solution</td>
			<td>TexWipe</td>
			<td>TX684</td>
			<td></td>
		</tr>
	</tbody>
</table>

body composition and metabolic caging analysis in high fat fed mice

身体成分的改变 (脂肪或精益质量), 代谢参数, 如全身耗氧量, 能量消耗, 和基质的利用, 以及食物摄取和体力活动等行为可以提供重要的信息, 关于疾病的潜在机制。考虑到身体成分和新陈代谢对肥胖的发展及其后继后遗症的重要性, 必须在临床前期研究中对这些参数进行准确的测量。在过去几十年中, 技术的进步使人们有可能以无侵入性和纵向的方式在啮齿动物模型中得出这些措施。因此, 这些代谢措施在评估基因操作 (例如挖空或转基因小鼠、病毒性击倒或基因过度表达)、实验药物/复合筛查和膳食的反应时证明是有用的,行为或身体活动干预。在这里, 我们描述了用于测量身体成分和代谢参数使用动物监测系统在喂食和高脂肪饲料喂养小鼠的协议。

在图 3中看到的结果显示, 在高脂肪喂养时, 身体成分参数的典型变化是通过 EchoMRI 测量的。在基线中, 任何测量参数 (图 3A-f) 都没有差别。然而, 在仅仅1周的高脂肪喂养后, 风组的体重、脂肪质量和脂肪质量百分比显著增加 (图 3A、B、D)。在3周的饮食干预中, 这两组人?...

Watch this Scientific Journal Video about Body Composition and Metabolic Caging Analysis in High Fat Fed Mice at JoVE.com

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

高脂喂养小鼠体内成分及代谢锁定分析

本协议描述了使用人体成分分析仪和代谢动物监测系统来表征小鼠体内成分和代谢参数。以高脂喂养诱发肥胖模型为例, 为这些技术的应用提供借鉴。

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

Research

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Medicine

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

在自觉的，奔放的小鼠正常血糖高胰岛素 - 夹钳

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold ...

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Protocol for Long Duration Whole Body Hyperthermia in Mice

持续时间较长的小鼠全身热疗协议

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

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When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

解剖的小鼠胰腺的组织学分析与代谢谱

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Semi-automated Analysis of Mouse Skeletal Muscle Morphology and Fiber-type Composition

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For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of ...

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test OGTT and Insulin Tolerance Test ITT

口服葡萄糖耐受试验 (OGTT) 和胰岛素耐受试验 (ITT) 对高脂饮食喂养小鼠体内葡萄糖代谢的影响研究 (英文)

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to ...

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis (NASH) Resolution

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis NASH Resolution

在小鼠中纳入可存活的肝脏活检程序评估非酒精性脂肪性肝炎 (NASH) 的解决方法

Clinical trials assessing therapies for the treatment of non-alcoholic steatohepatitis (NASH) involve a baseline and end of study liver biopsy, and ...

Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice

平滑肌细胞近系追踪小鼠晚期动脉粥样硬化病变细胞成分的定量分析

Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel ...

Vinyl Chloride and High-Fat Diet as a Model of Environment and Obesity Interaction

氯乙烯和高脂肪饮食作为环境与肥胖相互作用的典范

Vinyl chloride (VC), an abundant environmental contaminant, causes steatohepatitis at high levels, but is considered safe at lower levels. Although ...

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使用切片-O-马蒂奇和霍罗斯进行身体成分分析的细分和线性测量

Body composition is associated with risk of disease progression and treatment complications in a variety of conditions. Therefore, quantification of ...