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Current protocols in mouse biology最新文献

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Genomic Mutation Identification in Mice Using Illumina Sequencing and Linux-Based Computational Methods 利用Illumina测序和基于Linux的计算方法鉴定小鼠基因组突变
Current protocols in mouse biology Pub Date : 2019-07-15 DOI: 10.1002/cpmo.64
John A. Williams, George Powell, Ann-Marie Mallon, Michelle M. Simon
{"title":"Genomic Mutation Identification in Mice Using Illumina Sequencing and Linux-Based Computational Methods","authors":"John A. Williams,&nbsp;George Powell,&nbsp;Ann-Marie Mallon,&nbsp;Michelle M. Simon","doi":"10.1002/cpmo.64","DOIUrl":"10.1002/cpmo.64","url":null,"abstract":"<p>Genetically modified mice are an essential tool for modeling disease-causing mechanisms and discovering gene function. SNP genotyping was traditionally used to associate candidate regions with traits in the mouse, but failed to reveal novel variants without further targeted sequencing. Using a robust set of computational protocols, we present a platform to enable scientists to detect variants arising from whole-genome and exome sequencing experiments. This article guides researchers on aligning reads to the mouse genome, quality-assurance strategies, mutation discovery, comparing mutations to previously discovered mouse SNPs, and the annotation of novel variants, in order to predict mutation consequences on the protein level. Challenges unique to the mouse are discussed, and two protocols use self-contained containers to maintain version control and allow users to adapt our approach to new techniques by upgrading container versions. Our protocols are suited for servers or office workstations and are usable by non-bioinformatics specialists. © 2019 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.64","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41820641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Issue Information TOC 发布信息TOC
Current protocols in mouse biology Pub Date : 2019-06-19 DOI: 10.1002/cpmo.53
{"title":"Issue Information TOC","authors":"","doi":"10.1002/cpmo.53","DOIUrl":"10.1002/cpmo.53","url":null,"abstract":"<p><b>Cover</b>: Image shows an embryonic day 18.5 (E18.5) mouse embryo imaged with three-dimensional iodine-contrast micro-computed tomography (microCT). The 3D volume and individual section views were rendered with CTVox (Bruker) and the final image was assembled in Photoshop CC (Adobe). See Hsu et al. (https://doi.org/10.1002/cpmo.63).\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.53","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46406056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
High Resolution Imaging of Mouse Embryos and Neonates with X-Ray Micro-Computed Tomography. 用x射线微计算机断层扫描技术对小鼠胚胎和新生儿进行高分辨率成像。
Current protocols in mouse biology Pub Date : 2019-06-01 Epub Date: 2019-06-13 DOI: 10.1002/cpmo.63
Chih-Wei Hsu, Sowmya Kalaga, Uchechukwu Akoma, Tara L Rasmussen, Audrey E Christiansen, Mary E Dickinson
{"title":"High Resolution Imaging of Mouse Embryos and Neonates with X-Ray Micro-Computed Tomography.","authors":"Chih-Wei Hsu,&nbsp;Sowmya Kalaga,&nbsp;Uchechukwu Akoma,&nbsp;Tara L Rasmussen,&nbsp;Audrey E Christiansen,&nbsp;Mary E Dickinson","doi":"10.1002/cpmo.63","DOIUrl":"https://doi.org/10.1002/cpmo.63","url":null,"abstract":"<p><p>Iodine-contrast micro-computed tomography (microCT) 3D imaging provides a non-destructive and high-throughput platform for studying mouse embryo and neonate development. Here we provide protocols on preparing mouse embryos and neonates between embryonic day 8.5 (E8.5) to postnatal day 4 (P4) for iodine-contrast microCT imaging. With the implementation of the STABILITY method to create a polymer-tissue hybrid structure, we have demonstrated that not only is soft tissue shrinkage minimized but also the minimum required time for soft tissue staining with iodine is decreased, especially for E18.5 to P4 samples. In addition, we also provide a protocol on using commercially available X-CLARITY<sup>TM</sup> hydrogel solution to create the similar polymer-tissue hybrid structure on delicate early post-implantation stage (E8.5 to E14.5) embryos. With its simple sample staining and mounting processes, this protocol is easy to adopt and implement for most of the commercially available, stand-alone microCT systems in order to study mouse development between early post-implantation to early postnatal stages. © 2019 by John Wiley & Sons, Inc.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 2","pages":"e63"},"PeriodicalIF":0.0,"publicationDate":"2019-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.63","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37330802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 9
Ultrasound-Guided Approaches to Improve Orthotopic Mouse Xenograft Models for Hepatocellular Carcinoma 超声引导下改善肝癌原位小鼠异种移植模型的研究
Current protocols in mouse biology Pub Date : 2019-05-30 DOI: 10.1002/cpmo.62
Ghina Bou About, Emilie Thiebault, Marie Wattenhofer-Donzé, Hugues Jacobs, Alain Guimond, Tania Sorg, Eric Robinet, Thomas F. Baumert, Laurent Monassier, Yann Herault
{"title":"Ultrasound-Guided Approaches to Improve Orthotopic Mouse Xenograft Models for Hepatocellular Carcinoma","authors":"Ghina Bou About,&nbsp;Emilie Thiebault,&nbsp;Marie Wattenhofer-Donzé,&nbsp;Hugues Jacobs,&nbsp;Alain Guimond,&nbsp;Tania Sorg,&nbsp;Eric Robinet,&nbsp;Thomas F. Baumert,&nbsp;Laurent Monassier,&nbsp;Yann Herault","doi":"10.1002/cpmo.62","DOIUrl":"10.1002/cpmo.62","url":null,"abstract":"<p>Hepatocellular carcinoma (HCC) is the second leading cause of cancer death worldwide. While curative approaches for early stage HCC exist, effective treatment options for advanced HCC are lacking. Furthermore, there are no efficient chemopreventive strategies to limit HCC development once cirrhosis is established. One challenge for drug development is unsatisfactory animal models. In this article, we describe an orthotopic xenograft mouse model of human liver cancer cell lines through image-guided injection into the liver. This technique provides a less invasive yet highly efficient approach to engraft human HCC into mouse liver. Similarly, image-guided injections are used to deliver chemotherapeutics locally, enabling reduction in potential systemic adverse effects, while reducing the required dose for a therapeutic effect. In summary, this image-guided strategy provides a novel and convenient approach to improve current HCC mouse models. © 2019 The Authors. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.62","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37292220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
Assessing Blood Clotting and Coagulation Factors in Mice 小鼠凝血和凝血因子的测定
Current protocols in mouse biology Pub Date : 2019-03-15 DOI: 10.1002/cpmo.61
Marisa A. Brake, Lacramioara Ivanciu, Susan A. Maroney, Nicolas D. Martinez, Alan E. Mast, Randal J. Westrick
{"title":"Assessing Blood Clotting and Coagulation Factors in Mice","authors":"Marisa A. Brake,&nbsp;Lacramioara Ivanciu,&nbsp;Susan A. Maroney,&nbsp;Nicolas D. Martinez,&nbsp;Alan E. Mast,&nbsp;Randal J. Westrick","doi":"10.1002/cpmo.61","DOIUrl":"10.1002/cpmo.61","url":null,"abstract":"<p>The mammalian blood coagulation system was designed to restrict blood loss due to injury as well as keep the blood fluid within the blood vessels of the organism. Blood coagulation activity in inbred mouse strains varies widely among strains, suggesting that many genomic variants affect hemostasis. Some of these molecules have been discovered and characterized; however, many are still unknown. Genetically modified mouse technologies are providing a plethora of new mouse models for investigating the regulation of blood coagulation. Here we provide a protocol for the tail bleeding time as a primary assessment of in vivo blood coagulation, as well as in vitro methods such as the prothrombin time, activated partial thromboplastin time, and thrombin generation assay. We also provide protocols for the assessment of the activities of specific known factors involved in blood coagulation. © 2019 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.61","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37221247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 19
Issue Information TOC 发布信息TOC
Current protocols in mouse biology Pub Date : 2019-03-05 DOI: 10.1002/cpmo.52
{"title":"Issue Information TOC","authors":"","doi":"10.1002/cpmo.52","DOIUrl":"https://doi.org/10.1002/cpmo.52","url":null,"abstract":"<p><b>Cover</b>: In Lange et al. (https://doi.org/10.1002/cpmo.59) image shows cecal and colonic fluorescent micrographs from representative GF mice and non-GF control mice housed in an IVC operated in containment mode for 4 weeks. Colon enterocytes were stained with DAPI, and bacterial cells were stained with the bacterial probe EUB338 tagged with an orange fluorescent dye (Alexa 555). <b>(A,B)</b> Cecum <b>(A)</b> and proximal colon <b>(B)</b> from a representative GF mouse showing intestinal digesta within the lumen (asterisks), but no observable bacterial cells. <b>(C,D)</b> Cecum <b>(C)</b> and proximal colon <b>(D)</b> from a representative non-GF control mouse showing abundant bacterial cells within the lumen (arrows). Scale bars, 200 µm.\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.52","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"137486409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Use of Isotope Tracers to Assess Lipid Absorption in Conscious Lymph Fistula Mice 利用同位素示踪剂评估有意识淋巴瘘小鼠的脂质吸收
Current protocols in mouse biology Pub Date : 2019-02-23 DOI: 10.1002/cpmo.60
Chih-Wei Ko, Jie Qu, Min Liu, Dennis D. Black, Patrick Tso
{"title":"Use of Isotope Tracers to Assess Lipid Absorption in Conscious Lymph Fistula Mice","authors":"Chih-Wei Ko,&nbsp;Jie Qu,&nbsp;Min Liu,&nbsp;Dennis D. Black,&nbsp;Patrick Tso","doi":"10.1002/cpmo.60","DOIUrl":"10.1002/cpmo.60","url":null,"abstract":"<p>This protocol provides a comprehensive reference for the evolution of the lymph fistula model, the mechanism of lipid absorption, the detailed procedure for studying lipid absorption using the lymph fistula model, the interpretation of the results, and consideration of the experimental design. The lymph fistula model is an approach to assess the concentration and rate of a range of molecules transported by the lymph by cannulating lymph duct in animals. In this protocol, mice first undergo surgery with the implantation of cannulae in the duodenum and mesenteric lymph duct and are allowed to recover overnight in Bollman restraining cages housed in a temperature-regulated environment. To study <i>in vivo</i> lipid absorption, a lipid emulsion is prepared with labeled tracers, including [<sup>3</sup>H]-triolein and [<sup>14</sup>C]-cholesterol. On the day of the experiment, mice are continuously infused with lipid emulsion via the duodenum for 6 hr, and lymph is usually collected hourly. At the end of the study, gastrointestinal segments and their luminal contents are collected separately for determination of the digestion, uptake, and transport of exogenous lipids. © 2019 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.60","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36996501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Housing Gnotobiotic Mice in Conventional Animal Facilities 在传统动物设施中饲养非生物小鼠
Current protocols in mouse biology Pub Date : 2019-01-15 DOI: 10.1002/cpmo.59
Máximo E. Lange, Richard R. E. Uwiera, G. Douglas Inglis
{"title":"Housing Gnotobiotic Mice in Conventional Animal Facilities","authors":"Máximo E. Lange,&nbsp;Richard R. E. Uwiera,&nbsp;G. Douglas Inglis","doi":"10.1002/cpmo.59","DOIUrl":"10.1002/cpmo.59","url":null,"abstract":"<p>Gnotobiotic mice are an established, robust model utilized in current research to study host-microbiota interactions. For years isolators have been used to rear germ-free and gnotobiotic mice. However, isolators can be costly and the segregation of treatments within the same isolator is problematic. Recently, methodologies for housing germ-free mice in specially designed individually ventilated cages (IVCs) operated under barrier mode have been developed; however, this equipment is costly and its operation in barrier mode for research involving germ-free mice and pathogens is not permissible under modern biosafety and biosecurity regulations. This article describes a method to house germ-free mice in a commonly available conventional IVC system operated under containment mode. This technique allows researchers to maintain the germ-free or gnotobiotic status of mice tested up to 4 weeks with weekly handling while working with pathogens using each IVC as a separate experimental unit. © 2019 Her Majesty the Queen in Right of Canada.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.59","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36855490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 9
Issue Information TOC 发布信息TOC
Current protocols in mouse biology Pub Date : 2018-12-05 DOI: 10.1002/cpmo.51
{"title":"Issue Information TOC","authors":"","doi":"10.1002/cpmo.51","DOIUrl":"10.1002/cpmo.51","url":null,"abstract":"","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"8 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.51","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43711026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Adeno-Associated Virus Production, Purification, and Titering 腺相关病毒的产生、纯化和滴度测定
Current protocols in mouse biology Pub Date : 2018-11-29 DOI: 10.1002/cpmo.56
Yong Hong Chen, Megan S. Keiser, Beverly L. Davidson
{"title":"Adeno-Associated Virus Production, Purification, and Titering","authors":"Yong Hong Chen,&nbsp;Megan S. Keiser,&nbsp;Beverly L. Davidson","doi":"10.1002/cpmo.56","DOIUrl":"10.1002/cpmo.56","url":null,"abstract":"<p>Adeno-associated virus (AAV) vectors are exemplary tools for studying gene function <i>in vivo</i> and are particularly favorable for transferring genes of interest into brain tissues. They have shown great promise as a gene therapy vector for preclinical and clinical applications. However, the ability to use this tool is often hampered because the viruses themselves are not readily available. Many methods have been developed for AAV production. Here, we describe a simple method for small- to medium-scale (10<sup>12</sup>-10<sup>13</sup> viral particles) production of AAV based on Polyethylenimine Max (PEI Max)–mediated triple transfection of HEK 293 cells and purification with iodixanol gradient ultracentrifugation. These methods will provide users with ample material of sufficient quality for performing <i>in vivo</i> gene transfer. © 2018 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":37980,"journal":{"name":"Current protocols in mouse biology","volume":"8 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmo.56","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36733164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 16
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