此视频发布成为可能，资金由美国国家糖尿病，消化道和肾脏疾病研究所（NIDDK）的计划项目资助计划"病理学的肠道系统"DK 68055。克里斯托弗·T. Creedon罗切斯特公立学校"营商友导"计划的支持。 我们感谢加里·斯托尔兹的技术援助，秘书服务的克里斯蒂Zodrow女士和道格拉斯博士贝尔公司（Los Gatos的研究山景，CA）。

1。对小鼠的培训和习惯化<ol><li>分析前，把所有的小鼠在测试室恒定气流的2-4小时，以习惯于他们的测试条件。这将显着降低压力水平，否则可能会导致异常的检测胃排空延迟。治疗小鼠犹如胃排空实验相同的方式运行。准备蛋黄（见下文），没有加入辛酸和饲料0.2克至每一个鼠标。 </li><li>重复这个过程，直到老鼠足够的训练（一般为2-3倍）。通常很容易将小鼠驯化，只要保持相同的环境条件。 </li></ol> 注:非习小鼠的继续走动，约1小时后转移到腔，大便和小便频繁，而习惯于的小鼠迅速解决他们在新的环境和休息悄悄。 没有（TE）:在实验过程中:监控动物的迹象，如过量排尿，排便，缺乏兴趣吃鸡蛋的习惯损失。如果是这样的情况下，考虑在一个空室重新仍属获得胃排空数据的前1-2次。一致性是非常重要的，而做这个实验。每次做的事情完全一样的方式是只有这样，才能得到可靠的和可重复的结果。这包括给予治疗（ 如胰岛素）每天在相同的时间，而不是分离的小鼠的笼子里的队友，除非绝对必要，空腹的老鼠，并在同一时间开始胃排空试验，并以同样的方式处理的小鼠。 2。同位素制备含试餐<ol><li>开始在50ml falcon试管中称量5克蛋黄。重复这些步骤，每一个实验一天的时间准备一顿一个新的测试。 </li><li>加入10μl的辛酸与concentr大力功能明显下降，2μL/克到50毫升Falcon管中含有鸡蛋和混合1分钟，用锅铲在猎鹰管。 </li><li>鸡蛋然后转移到玻璃烧杯中，并加热超过本生灯直到它凝固，其一致性是合适的，以使小球。这通常需要约30秒。 </li></ol> 注:蛋黄球重达0.2克每鼠标。这是很重要的，在所有的小鼠中的累积剂量保持恒定。 3。开始实验<ol><li>一旦训练和准备，对胃的排空，快速的小鼠的金属"网底"禁食架上，以防止coprophagia过夜（12小时）。确保他们有免费的饮用水。由于糖尿病小鼠中使用在目前的实验中，它们不应该被禁食16小时以上。 </li><li>从胃排空室开始。使用洁净室和盖已风干。此外，任何的分析仪或CO 2的空气供给管连接腔室应防潮自由水可以由分析仪读出的信号干扰</li><li>连接腔室，以提供一个恒定的空气流的入口管。然后将出口管机室。关闭管和打开的气流。 </li></ol> 注:应用极少量的凡士林结束时盖盖，使他们容易和安全地密封。这是需要收集所有的小鼠产生的二氧化碳密封。 4。实验步骤<ol><li>通过衡量每个鼠标启动。体重是衡量其持续良好的健康状况。在适当的腔，然后将每个鼠标。这当然是重要的有空气流入到腔室，在这个时候。 </li><li>开始测量，使小鼠腔前调整来适应环境的空气。 </li><li>一旦小鼠出现平静，这可能需要几分钟的时间，每个鼠标室调节空气流量。这可能是不同的各小鼠。通常情况下，空气流被调整在实验开始时，以确保，呼出的CO 2达到正在使用的任何设备可检测的水平，以确保该水平保持足够低，以确保健康的空气周转。我们使用初始CO 2浓度百万分之1000至1500。 </li><li>如果有困难的调整，检查是否有漏气现象。然后重复这个过程，每个室，观看新一轮的测量，看是否调整气流已经纠正了CO 2。重要的是要获得稳定的基线，读取之前喂养的小鼠。自校准功能，我们使用一台机器。如果这不是应检查的情况下校准。 </li><li>当做到这一点，管理的鸡蛋餐第一个鼠标和记录的时间，每个鼠标接收的食物。 </li><li>我们运行该过程4小时，以获得足够的值，用于装配的13 CO 2富集曲线为每个鼠标。检查对小鼠每30-60分钟，以确保的CO 2水平仍然是安全的小鼠。 </li><li>准备新的箱子，里面装有食品年底前的测试，所以老鼠可以吃后立即测试结束。 </li></ol>

<ol>
	<li>Yeung, C. K., McCurrie, 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=A+simple+method+to+investigate+the+inhibitory+effects+of+drugs+on+gastric+emptying+in+the+mouse+in+vivo.">A simple method to investigate the inhibitory effects of drugs on gastric emptying in the mouse in vivo.</a> J. Pharmacol. Toxicol. Methods. 45, 235-240 (2001).</li><li>Osinski, M. A., Seifert, T. R., Cox, B. F., Gintant, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+method+of+evaluation+of+drug-evoked+changes+in+gastric+emptying+in+mice.">An improved method of evaluation of drug-evoked changes in gastric emptying in mice.</a> J. Pharmacol. Toxicol. Methods. 47, 115-120 (2002).</li><li>Ghoos, Y. F., 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=Measurement+of+gastric+emptying+rate+of+solids+by+means+of+a+carbon-labeled+octanoic+acid+breath+test.">Measurement of gastric emptying rate of solids by means of a carbon-labeled octanoic acid breath test.</a> Gastroenterology. 104, 1640-1647 (1993).</li><li>Symonds, E., Butler, R., Omari, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+breath+tests+can+detect+alterations+in+gastric+emptying+in+the+mouse.">Noninvasive breath tests can detect alterations in gastric emptying in the mouse.</a> Eur. J. Clin. Invest. 32, 341-344 (2002).</li><li>Symonds, E. L., Butler, R. N., Omari, T. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+gastric+emptying+in+the+mouse+using+the+[13C]-octanoic+acid+breath+test.">Assessment of gastric emptying in the mouse using the [13C]-octanoic acid breath test.</a> Clin. Exp. Pharmacol. Physiol. 27, 671-675 (2000).</li><li>Verhulst, 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=Role+of+ghrelin+in+the+relationship+between+hyperphagia+and+accelerated+gastric+emptying+in+diabetic+mice.">Role of ghrelin in the relationship between hyperphagia and accelerated gastric emptying in diabetic mice.</a> Gastroenterology. 135, 1267-1276 (2008).</li><li>Schoonjans, 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=The+13C-octanoic+acid+breath+test:+validation+of+a+new+noninvasive+method+of+measuring+gastric+emptying+in+rats.">The 13C-octanoic acid breath test: validation of a new noninvasive method of measuring gastric emptying in rats.</a> Neurogastroenterol. Motil. 14, 287-293 (2002).</li><li>Choi, K. 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=Determination+of+gastric+emptying+in+nonobese+diabetic+mice.">Determination of gastric emptying in nonobese diabetic mice.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 293, 1039-1045 (2007).</li><li>Atkinson, M. A., Leiter, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+NOD+mouse+model+of+type+1+diabetes:+as+good+as+it+gets?.">The NOD mouse model of type 1 diabetes: as good as it gets?.</a> Nat. Med. 5, 601-604 (1999).</li><li>Camilleri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+practice.+Diabetic+gastroparesis.">Clinical practice. Diabetic gastroparesis.</a> N. Engl. J. Med. 356, 820-829 (2007).</li><li>Choi, K. 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=Heme+oxygenase-1+protects+interstitial+cells+of+Cajal+from+oxidative+stress+and+reverses+diabetic+gastroparesis.">Heme oxygenase-1 protects interstitial cells of Cajal from oxidative stress and reverses diabetic gastroparesis.</a> Gastroenterology. 135, 2055-2064 (2008).</li><li>Kashyap, P. 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=Carbon+monoxide+reverses+diabetic+gastroparesis+in+NOD+mice.">Carbon monoxide reverses diabetic gastroparesis in NOD mice.</a> Am. J. Physiol. GI. G298, G1013-G1019 (2010).</li></ol>

作者宣称，他们没有竞争的金融利益。

本文所描述的技术允许重复的和非侵入性的， 在体内测量的固体小鼠胃排空的影响。这种系统具有的优点在于，将动物没有在测量室中的限制，，使他们可自由移动和转动。由于这是一个陌生的环境中，这些老鼠仍然需要进行培训和熟悉的试验室，以防止压力的影响胃排空。在一般情况下，我们假定胃排空数据是可靠的，如果连续胃排空试验帧内小鼠之间的变异小于10％。 <p class="jove_co...

这手稿描述的技术和方法上的考虑，在非侵入性测量小鼠胃排空的影响。通过这里描述的协议，研究人员可以可靠和可重复的在胃的排空，由于疾病发展变化，研究药物制剂的影响，对胃排空及胃的排空，以治疗基础疾病或缺陷6,8， 11,12。在以前的出版物中，应用13 C辛酸呼气测试，被证明是一个有用的方法，测量胃排空的人类和动物3,8。本文详细介绍了必要的程序，以获得可靠的数据，在6〜8个月，有必要对糖尿病小鼠胃排空的纵向研究。遵循这个协议相比此前公布的方法的优点是，研究者可以放心的数据obtained将是可靠和可重复性。此外，这里描述的自动化系统，用于收集和分析的气体样品的动物的数量增加，同时在一项研究中，可以遵循的。总之，本文的目标是要找出保持成瘾的老鼠的测试，得到的结果，减少变异的关键因素。 小鼠体内检测胃排空，禁食过夜，投入透明的塑料试验箱，恒定气流。基线后小老鼠就习惯于管，呼出的13 CO 2级确定和气流相应的调整。接下来，我们测试管理用餐组成的蛋黄混合，用13 C-标记的辛酸。因为禁食小鼠和训练，他们一般在2分钟内吃的试验餐。施用的辛酸还没有在胃中吸收，但将会采取在十二指肠d将浏览在肝脏中代谢成13 CO 2，这是释放和呼出，从而在周围的空气中的13 CO 2的富集。收集空气样品，以确定的时间间隔，并且由碳同位素分析仪分析。在这整个过程的限速步骤是胃的排空和肺排泄的13 CO 2直接对应的标记餐的胃排空。 <img src="/files/ftp_upload/50301/50301fig1.jpg" alt="图1" fo:content-width="6in" fo:src="/files/ftp_upload/50301/50301fig1highres.jpg"/> 图1。胃的排空装置的示意图。禁食一夜后，小鼠被放置在透明的商会，使他们能够自由地移动和转动。入口管允许新鲜的和恒定的空气流入和一个出口导致的同位素分析仪测量的13 C-12C比值的呼出一口气。该室也有一个中央端口提供的食物中含有[13 C]辛酸。

<table border="1"><tbody><tr><td> NAME </td><td> 公司 </td><td> CAT NUMBER </td><td> 评论 </td></tr><tr><td>一位消息人士的恒定供气，流量和组成</td><td>中央供气的研究设施</td><td></td><td></td></tr><tr><td> 130毫升取样室，有进风口，出风口，与食品管理局开幕 塑料管的空气供给</td><td>在内部构建</td><td></td><td></td></tr><tr><td>辛酸</td><td>剑桥同位素实验室（安多弗，MA） </td><td> CLM-293-1 </td><td></td></tr><tr><td>要准备的鸡蛋饭: <ul><li>小烧杯</li><li> 50毫升的塑胶管</li><li>本生灯</li><li>鸡蛋</li><li>抹刀</li></ul></td><td>任何供应商</td><td></td><td>尝试是一致的鸡蛋供应商，因为鸡蛋的营养成分和适口性的影响摄食和胃排空的餐</td></tr><tr><td>二氧化碳同位素分析仪</td><td> Los Gatos的研究公司（加州Mountain View） </td><td></td><td></td></tr></tbody></table>

assessment of gastric emptying in non-obese diabetic mice using a [13c]-octanoic acid breath test

在小鼠的胃排空研究一直局限于由无法遵循胃排空以来的变化在同一动物的最常用的技术需要杀死动物和死后恢复，用餐1,2。这种方法可以防止纵向研究，以确定胃排空的变化，随着年龄的增长和疾病的进展。目前常用的[13 C]-辛酸呼气试验对人类3被修改用于在小鼠4-6和大鼠7我们以前表明，这个测试是可靠和胃排空的变化响应于在对药物的反应，以及在糖尿病患者病情恶化。在这段视频中演示的原理和实际执行这个修改后的测试进行了解释。在以前的研究中，NOD LTJ小鼠，1型糖尿病9的典范。这些小鼠的发展一定比例的胃轻瘫的症状，一期糖尿病并发症ES特点是胃排空延迟，无机械性梗阻，胃10。 本文演示了如何训练的老鼠进行测试，如何准备试验餐，并获得4小时胃排空的数据，如何分析所获得的数据。在本研究中所使用的碳同位素分析仪是适合的空气样品的自动采样从多达12只小鼠在同一时间。这种技术允许的纵向后续胃排空组小鼠与糖尿病或其他长期疾病。

从三个不同的小鼠设置，典型的数据在图2中所示。从小鼠胃排空正常的黑色图形表示的数据点。它示出的13 C]是表示为每小时的给药剂量的百分比表示为时间的函数的呼出的空气中回收的馏分。蓝色曲线是鼠标加速胃排空的T半值的40分钟，而红色曲线是从小鼠胃排空延迟与一个T的一半价值168分钟。建立在我们的实验室7，正常非糖尿病NOD小鼠年龄胃半排空时间为9-15周?...

Watch this Scientific Journal Video about Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test at JoVE.com

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

与一种非侵入性测定胃排空[<sup&gt; 13</sup&gt; C-辛酸呼气试验跟踪女性NOD LTJ小鼠的胃轻瘫。

使用的非肥胖型糖尿病小鼠胃排空的评估[<sup&gt; 13</sup&gt; C-辛酸呼气试验

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

assessment-gastric-emptying-non-obese-diabetic-mice-using-13c

Research

JoVE Journal

Medicine

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

13.5K 次观看.  Mayo Clinic. 与一种非侵入性测定胃排空[ 13 C-辛酸呼气试验跟踪女性NOD LTJ小鼠的胃轻瘫。

视频: Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Utilizing 18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer

A hallmark of advanced tumors is a switch to aerobic glycolysis that is readily measured by [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography (18F-FDG PET) imaging. Co-mutations in the KRAS proto-oncogene and the LKB1 tumor suppressor gene are frequent events in lung cancer that drive hypermetabolic, glycolytic tumor growth. A critical pathway regulating the growth and metabolism of these tumors is the mechanistic target of the rapamycin (mTOR) pathway, which can be effectively targeted using selective catalytic mTOR kinase inhibitors. The mTOR inhibitor MLN0128 suppresses glycolysis in mice bearing tumors with Kras and Lkb1 co-mutations, referred to as KL mice. The therapy response in KL mice is first measured by 18F-FDG PET and computed tomography (CT) imaging before and after the delivery of MLN0128. By utilizing 18F-FDG PET/CT, researchers are able to measure dynamic changes in the glucose metabolism in genetically engineered mouse models (GEMMs) of lung cancer following a therapeutic intervention with targeted therapies. This is followed by ex vivo autoradiography and a quantitative immunohistochemical (qIHC) analysis using morphometric software. The use of qIHC enables the detection and quantification of distinct changes in the biomarker profiles following treatment as well as the characterization of distinct tumor pathologies. The coupling of PET imaging to quantitative histology is an effective strategy to identify metabolic and therapeutic responses in vivo in mouse models of disease.

Utilizing 18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer

In this protocol, we describe how to utilize [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography (18F-FDG PET/CT) imaging to measure the tumor metabolic response to the targeted therapy MLN0128 in a Kras/Lkb1 mutant mouse model of lung cancer and coupled imaging with high resolution ex vivo autoradiography and quantitative histology.

利用18f-葡萄糖 PET/CT 成像和定量组织学测量肺癌小鼠模型葡萄糖代谢的动态变化

A hallmark of advanced tumors is a switch to aerobic glycolysis that is readily measured by [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography ...

科研

癌症研究

Hyponeophagia: A Measure of Anxiety in the Mouse

Before the present day, when fast-acting and potent rodenticides such as alpha-chloralose were not yet in use, the work of pest controllers was often hampered by a phenomenon known as "bait shyness". Mice and rats cannot vomit, due to the tightness of the cardiac sphincter of the stomach, so to overcome the problem of potential food toxicity they have evolved a strategy of first ingesting only very small amounts of novel substances. The amounts ingested then gradually increase until the animal has determined whether the substance is safe and nutritious. So the old rat-catchers would first put a palatable substance such as oatmeal, which was to be the vehicle for the toxin, in the infested area. Only when large amounts were being readily consumed would they then add the poison, in amounts calculated not to affect the taste of the vehicle. The poisoned bait, which the animals were now readily eating in large amounts, would then swiftly perform its function.
Bait shyness is now used in the behavioural laboratory as a way of measuring anxiety. A highly palatable but novel substance, such as sweet corn, nuts or sweetened condensed milk, is offered to the mice (or rats) in a novel situation, such as a new cage. The latency to consume a defined amount of the new food is then measured.
Robert M.J. Deacon can be reach at <a href="mailto:robert.deacon@psy.ox.ac.uk">robert.deacon@psy.ox.ac.uk</a>

Mice and rats, due to their innate cautiousness, are initially slow in consuming a novel food, particularly in a novel place. This hyponeophagia can readily be measured in the laboratory, even though laboratory animals are much less anxious than their wild counterparts

Hyponeophagia:在小鼠的焦虑测量

Before the present day, when fast-acting and potent rodenticides such as alpha-chloralose were not yet in use, the work of pest controllers was often ...

神经科学

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an area of productive research with over 1,400 published animal studies in just the last 5 years. Numerous techniques have been developed for quantifying smooth muscle activity of the stomach, small intestine, and colon. In vitro and ex vivo techniques offer powerful tools for mechanistic studies of GI function, but outside the context of the integrated systems inherent to an intact organism. Typically, measuring in vivo smooth muscle contractions of the stomach has involved an anesthetized preparation coupled with the introduction of a surgically placed pressure sensor, a static pressure load such as a mildly inflated balloon or by distending the stomach with fluid under barostatically-controlled feedback. Yet many of these approaches present unique disadvantages regarding both the interpretation of results as well as applicability for in vivo use in conscious experimental animal models. The use of dual element strain gages that have been affixed to the serosal surface of the GI tract has offered numerous experimental advantages, which may continue to outweigh the disadvantages. Since these gages are not commercially available, this video presentation provides a detailed, step-by-step guide to the fabrication of the current design of these gages. The strain gage described in this protocol is a design for recording gastric motility in rats. This design has been modified for recording smooth muscle activity along the entire GI tract and requires only subtle variation in the overall fabrication. Representative data from the entire GI tract are included as well as discussion of analysis methods, data interpretation and presentation.

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

The in vivo measurement of smooth muscle contractions along the gastrointestinal tract of laboratory animals remains a powerful, though underutilized, technique. Flexible, dual element strain gages are not commercially available and require fabrication. This protocol describes the construction of reliable, inexpensive strain gages for acute or chronic implantation in rodents.

的微型双元应变计测量加工和植入<em&gt;体内</em&gt;胃肠收缩鼠害。

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an ...

生物工程

Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmonary Outcomes

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese individuals worldwide by 2030, thus representing a growing socioeconomic burden. Simultaneously, obesity-related comorbidities, including diabetes as well as heart and chronic lung diseases, are continuously on the rise. Although obesity has been associated with increased risk for asthma exacerbations, worsening of respiratory symptoms, and poor control, the functional role of obesity and perturbed metabolism in the pathogenesis of chronic lung disease is often underestimated, and underlying molecular mechanisms remain elusive. This article aims to present methods to assess the effect of obesity on metabolism, as well as lung structure and function. Here, we describe three techniques for mice studies: (1) assessment of intraperitoneal glucose tolerance (ipGTT) to analyze the effect of obesity on glucose metabolism; (2) measurement of airway resistance (Res) and respiratory system compliance (Cdyn) to analyze the effect of obesity on lung function; and (3) preparation and fixation of the lung for subsequent quantitative histological assessment. Obesity-related lung diseases are probably multifactorial, stemming from systemic inflammatory and metabolic dysregulation that potentially adversely influence lung function and the response to therapy. Therefore, a standardized methodology to study molecular mechanisms and the effect of novel treatments is essential.

The incidence of obesity is rising and increases the risk of chronic lung diseases. To establish the underlying mechanisms and preventive strategies, well-defined animal models are needed. Here, we provide three methods (glucose-tolerance-test, body plethysmography, and lung fixation) to study the effect of obesity on pulmonary outcomes in mice.

腹腔葡萄糖耐受试验、肺功能测定和肺固定研究肥胖和代谢障碍对肺部结局的影响

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese ...

免疫与感染

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is frequently associated with metabolic complications, such as type 2 diabetes, hypertension, and dyslipidemia, reducing life expectancy and quality. In the absence of any effective noninvasive treatments, bariatric surgery is a valuable therapeutic option for patients with morbid obesity (body mass index (BMI) &#62;40 kg/m2), leading to long-term, sustained weight loss and improvements in metabolic complications. However, the underlying cellular and molecular mechanisms sustaining the beneficial effects of bariatric surgery are not yet fully understood. Due to the numerous genetically-modified strains available, the mouse model is the most convenient animal model to explore the molecular mechanisms behind the pleiotropic beneficial effects of bariatric surgeries. Here, we detailed the optimized healthcare methods and surgical protocols in mice for the two most widely-used bariatric surgeries: the sleeve gastrectomy and the modified Roux-en-Y gastric bypass. Deciphering the molecular mechanisms underlying the therapeutic effects of bariatric surgeries offers the promise of identifying new therapeutics targets.

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

袖胃切除术的技术和改良的小鼠空肠Roux-en-Y胃旁路

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

医学

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

确定葡萄糖代谢动力学研究<sup&gt; 18</sup&gt; F-FDG微PET / CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

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

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to insulin-stimulated glucose uptake and a gross decompensation of systemic glucose metabolism. Despite considerable progress in the understanding of glucose metabolism, the molecular mechanisms of its regulation in health and disease remain under-investigated, while novel approaches to prevent and treat diabetes are urgently needed. Diet derived glucose stimulates the pancreatic secretion of insulin, which serves as the principal regulator of cellular anabolic processes during the fed-state and thus balances blood glucose levels to maintain systemic energy status. Chronic overfeeding triggers meta-inflammation, which leads to alterations in peripheral insulin receptor-associated signaling and thus reduces the sensitivity to insulin-mediated glucose disposal. These events ultimately result in elevated fasting glucose and insulin levels as well as a reduction in glucose tolerance, which in turn serve as important indicators of insulin resistance. Here, we present a protocol for the generation and metabolic characterization of high-fat diet (HFD)-fed mice as a frequently used model of diet-induced insulin resistance. We illustrate in detail the oral glucose tolerance test (OGTT), which monitors the peripheral disposal of an orally administered glucose load and insulin secretion over time. Additionally, we present a protocol for the insulin tolerance test (ITT) to monitor whole-body insulin action. Together, these methods and their downstream applications represent powerful tools to characterize the general metabolic phenotype of mice as well as to specifically assess alterations in glucose metabolism. They may be especially useful in the broad research field of insulin resistance, diabetes and obesity to provide a better understanding of pathogenesis as well as to test the effects of therapeutic interventions.

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

The current article describes the generation and metabolic characterization of high-fat diet-fed mice as a model of diet-induced insulin resistance and obesity. It further features detailed protocols to perform the oral glucose tolerance test and the insulin tolerance test, monitoring whole-body alterations of glucose metabolism in vivo.

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 ...

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

高脂喂养小鼠体内成分及代谢锁定分析

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

Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract metabolic abnormalities. So far, different types of IF models with varying durations of fasting and feeding periods have been documented. However, interpreting the outcomes is challenging, as many of these models involve multifactorial contributions from both time- and calorie-restriction strategies. For example, the alternate day fasting model, often used as a rodent IF regimen, can result in underfeeding, suggesting that health benefits from this intervention are likely mediated via both caloric restriction and fasting-refeeding cycles. Recently, it has been successfully demonstrated that 2:1 IF, comprising 1 day of fasting followed by 2 days of feeding, can provide protection against diet-induced obesity and metabolic improvements without a reduction in overall caloric intake. Presented here is a protocol of this isocaloric 2:1 IF intervention in mice. Also described is a pair-feeding (PF) protocol required to examine a mouse model with altered eating behaviors, such as hyperphagia. Using the 2:1 IF regimen, it is demonstrated that isocaloric IF leads to reduced body weight gain, improved glucose homeostasis, and elevated energy expenditure. Thus, this regimen may be useful to investigate the health impacts of IF on various disease conditions.

The current article describes a detailed protocol for isocaloric 2:1 intermittent fasting to protect and treat against obesity and impaired glucose metabolism in wild-type and ob/ob mice.

小鼠间间间禁食的代谢效应评估 2:1 间歇禁食

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract ...

生物学

<meta charset="utf8"></head><body>高脂饮食和 STZ 注射治疗糖尿病心肌病的小鼠模型

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.

<meta charset="utf8"></head><body>小鼠糖尿病心肌病模型的建模与评价

This protocol outlines a method for inducing diabetic cardiomyopathy through a combination of high-fat diet feeding and streptozotocin injection. This approach aims to provide a reliable framework for scientific investigation into diabetic cardiomyopathy and to explore potential avenues for clinical treatment applications.

小鼠糖尿病心肌病模型的建模与评价

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus ...

使用的非肥胖型糖尿病小鼠胃排空的评估[

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利用¹⁸f-葡萄糖 PET/CT 成像和定量组织学测量肺癌小鼠模型葡萄糖代谢的动态变化