Name: dissection of the mouse pancreas for histological analysis and metabolic profiling
Uploaded: 2017-08-19T14:00:00
Description: Watch this Scientific Journal Video about Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling at JoVE.com

麦承认支持这项工作从国立卫生研究院和国家癌症研究所资助编号-1R15CA152985 01A1。这个项目也得到了奖学金计划和迈阿密大学学者暑期的迈阿密大学博士学位本科机会迈阿密大学本科教学科研奖励计划。

在视频中进行，如下所述的程序已获机构动物护理和使用委员会 (IACUC) 在迈阿密大学。
 1.制备及刺激试验 <ol><li>建立两个不同的区域，为外科手术、 手术台和术后表。舞台这两个领域与所有材料和必要的用具。
	<ol><li>阶段的操作表排气罩下。把桌子布置与设备允许程序的连续和不受阻碍地性能的方式。</li>
		<li>建立一个术后的表在同一个房间里和附近的主表的操作。保持...

<ol>
	<li>Fesinmeyer, M. D., Austin, M. A., Li, C. I., De Roos, A. J., Bowen, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+Survival+by+Histologic+Type+of+Pancreatic+Cancer.">Differences in Survival by Histologic Type of Pancreatic Cancer.</a> Cancer Epidemiol Biomarkers Prev. 14 (7), 1766-1773 (2005).</li><li>Jackson-Grusby, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+cancer+in+mice.">Modeling cancer in mice.</a> Oncogene. 21 (35), 5504-5514 (2002).</li><li>Heid, I., 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=Early+requirement+of+Rac1+in+a+mouse+model+of+pancreatic+cancer.">Early requirement of Rac1 in a mouse model of pancreatic cancer.</a> Gastroenterology. 141 (2), 719-730 (2011).</li><li>Hingorani, S. 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=Preinvasive+and+invasive+ductal+pancreatic+cancer+and+its+early+detection+in+the+mouse.">Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse.</a> Cancer cell. 4 (6), 437-450 (2003).</li><li>Shi, 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=KRAS2+Mutations+in+Human+Pancreatic+Acinar-Ductal+Metaplastic+Lesions+Are+Limited+to+Those+with+PanIN+Implications+for+the+Human+Pancreatic+Cancer+Cell+of+Origin.">KRAS2 Mutations in Human Pancreatic Acinar-Ductal Metaplastic Lesions Are Limited to Those with PanIN Implications for the Human Pancreatic Cancer Cell of Origin.</a> Mol Cancer Res. 7 (2), 230-236 (2009).</li><li>LaConti, J. 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=Distinct+serum+metabolomics+profiles+associated+with+malignant+progression+in+the+KrasG12D+mouse+model+of+pancreatic+ductal+adenocarcinoma.">Distinct serum metabolomics profiles associated with malignant progression in the KrasG12D mouse model of pancreatic ductal adenocarcinoma.</a> BMC Genomics. 16 (1), 1-10 (2015).</li><li>Ludwig, M. 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=Surveying+the+serologic+proteome+in+a+tissue-specific+kras(G12D)+knockin+mouse+model+of+pancreatic+cancer.">Surveying the serologic proteome in a tissue-specific kras(G12D) knockin mouse model of pancreatic cancer.</a> Proteomics. 16 (3), 516-531 (2016).</li><li>Taylor, D. K., Mook, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+Waste+Anesthetic+Gas+Concentrations+Associated+with+the+Open-Drop+Method.">Isoflurane Waste Anesthetic Gas Concentrations Associated with the Open-Drop Method.</a> J Am Assoc Lab Anim Sci. 48 (1), 61-64 (2009).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Pharmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Li, D. S., Yuan, Y. H., Tu, H. J., Liang, Q. L., Dai, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+protocol+for+islet+isolation+from+mouse+pancreas.">A protocol for islet isolation from mouse pancreas.</a> Nature Protocols. 4 (11), 1649-1652 (2009).</li><li>Auer, 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=The+effects+of+frozen+tissue+storage+conditions+on+the+integrity+of+RNA+and+protein.">The effects of frozen tissue storage conditions on the integrity of RNA and protein.</a> Biotech Histochem. 89 (7), 518-528 (2014).</li><li>Ma, J., Leung, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limbic+System+Participates+in+Mediating+the+Effects+of+General+Anesthetics.">Limbic System Participates in Mediating the Effects of General Anesthetics.</a> Neuropsychopharmacology. 31, 1177-1192 (2006).</li><li>Ijichi, 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=Aggressive+pancreatic+ductal+adenocarcinoma+in+mice+caused+by+pancreas-specific+blockade+of+transforming+growth+factor-&#946;+signaling+in+cooperation+with+active+Kras+expression.">Aggressive pancreatic ductal adenocarcinoma in mice caused by pancreas-specific blockade of transforming growth factor-&#946; signaling in cooperation with active Kras expression.</a> Genes Dev. 20 (22), 3147-3160 (2006).</li><li>Abiatari, I., 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=Moesin-dependent+cytoskeleton+remodelling+is+associated+with+an+anaplastic+phenotype+of+pancreatic+cancer.">Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.</a> J Cell Mol Med. 14 (5), 1166-1179 (2010).</li><li>Goodpaster, A. M., Romick-Rosendale, L. E., Kennedy, M. 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+Statistical+significance+analysis+of+nuclear+magnetic+resonance-based+metabonomics+data.">a Statistical significance analysis of nuclear magnetic resonance-based metabonomics data.</a> Analytical biochemistry. 401, 134-143 (2010).</li><li>Yadav, D., 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=Idiopathic+Tumefactive+Chronic+Pancreatitis:+Clinical+Profile,+Histology,+and+Natural+History+After+Resection.">Idiopathic Tumefactive Chronic Pancreatitis: Clinical Profile, Histology, and Natural History After Resection.</a> Clin Gastroenterol Hepatol. 1 (2), 129-135 (2003).</li></ol>

重要的意义，现有的方法 虽然其他非正式的视频鼠标夹层动脉瘤存在，视频本文提供专业质量第一、 同行评审、 直观示范的所有详细的步骤所需的提取和收获的小鼠胰腺的解剖10.正在主要器官代谢活性和胰岛素的胰腺生产、 夹层和收获的胰腺允许保存的生理特性11。通过隔离胰腺，可能对样品进行未来的分析。这个程序允许从其他组织在相同?...

鼠标已成为人类胰腺癌1，2重要动物模型。在 Ptf1acre / +;LSL KrasG12D / +小鼠模型，大鼠肉瘤 (K-Ras) 癌基因激活只在胰腺，导致胰腺，胰腺上皮内肿瘤 (PanINs)，这一进展被称为癌前病变引发的克里斯汀对胰腺导管腺癌，通常所说的 PDACs3。这个鼠标模型系统提供了一个人类胰腺癌4，5，与 PanINs 出现在首五个月的生活和频繁地进展 PDAC 内的附加优势的最佳动物模型单 44，5，而胰腺癌最频繁地出现在人类身上 60-70 岁的年龄。
由 Ptf1a 淋巴结清扫术胰腺提取cre / +;LSL KrasG12D / +小鼠不同年龄允许详细纵向组织学检查癌症发展的胰腺，从最早的 PanIN 阶段通过进展到 PDAC3，4,5.在年龄从五到十五个月收获胰腺可也用于准备组织提取物表征在胰腺4代谢在从健康到病变组织的过渡过程中发生的全球变化6，7。
本文介绍了一个完整的视觉指南执行鼠标胰腺提取所需的步骤并提供指导方针的胰腺，进一步分析存储。本指南将同样宝贵，为个人进行研究其他胰腺的疾病，包括 I 型糖尿病，和应该是特别有用的学生和研究人员的新研究涉及收获的小鼠胰腺使用清扫术代谢轮廓分析或组织学分析。

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dissection of the mouse pancreas for histological analysis and metabolic profiling

我们一直在调查胰腺特异性转录因子、 1a-cre 重组酶;脂氧合酶-停止-脂氧合酶-克里斯汀大鼠肉瘤、 甘氨酸对天冬氨酸在 12 密码子 (Ptf1acre / +;LSL-KrasG12D / +) 鼠标应变作为人类胰腺癌模型。我们当前的研究的目标是找出新型代谢生物标记物的胰腺肿瘤。我们进行代谢过程的尿液、 粪便、 血液，和胰腺组织提取物，以及胰腺期癌的组织学分析。小鼠胰腺不是定义良好的实质器官，像人类，而是弥漫性分布的软组织不容易标识由个人熟悉鼠标内部解剖或有很少或没有表演经验的个人鼠标器官解剖。这篇文章的目的是提供一个详细的逐级直观示范指导新手在小鼠胰腺切除的解剖。这篇文章应该向学生和研究人员新需要收获的小鼠胰腺剥离术治疗代谢轮廓分析或组织学分析的研究尤为重要。

图 1显示的操作环境区的概述和图 2显示了岗位操作区域。虽然此设置提供最小数量的设备和分期，个人可以选择改变了这一点，最适合个人需要。议定书 》 应根据实验的具体需要进行优化。此过程以终止的一只老鼠，需要适当的狗狗会被安乐死9生活的方式进行。当研究者是准备好了时，鼠标放入麻醉室与异...

Watch this Scientific Journal Video about Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling at JoVE.com

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

本文视频提供了详细的演示了从一只老鼠成功删除胰腺解剖组织学分析和代谢过程所需的程序。

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 ...

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人类蛋白质组分析玻璃体元素剖析

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This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

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通过胰管结扎手术损伤的小鼠胰腺作为模型内分泌和外重新规划和扩散

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

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该做什么和不该做什么的肌肉冰冻切片的准备组织学分析

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of ...

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应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

超声心动图与小鼠心脏形态学的组织学检查

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

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The placenta is essential for the growth and development of mammalian embryos. For this reason, numerous genetic alterations and likely also environmental ...

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Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

利用隔离的完美大鼠小肠进行肠道激素分泌的机制

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