Discovery Strategies 2000
Emerging Technologies for Creating and Phenotyping New Mouse Models
This conference will present new advances in technologies, specifically directed toward creating and phenotyping genetically engineered mice. The conference will be presented in two sessions. The first session will be devoted to recent advances in molecular technologies in conditional mutagenesis, gene trap methods, ES cell mutagenesis and stem cell cloning. The second session will present new methods, technologies and equipment for phenotyping mouse models; expression profiling with microarrays, real time in vivo MRI, automated home cage behavior monitoring, and non-invasive measurement systems will all be presented.
Speaker List:
Gene Expression Profiling of Alopecia Areata: from mouse model to human disease
Joseph Carroll
Genetics Institute
To better understand the autoimmune disease alopecia areata, gene chip analysis was performed on mouse skin samples at various stages of the disease. Cluster analysis was used to group gene changes into families and this data was then compared with gene changes in chronic alopecia areata patients. Novel and known pathways in autoimmune disease from this model can now be compared to the human disease. This study represents a concise attempt to use gene expression monitoring to compare an animal model to a human disease in terms of mechanism and gene marker analysis.
The Virtual Microscope
Jose Galvez, MD
University of California, Davis
Continuing development of the virtual mouse as a teaching tool, highlighting virtual dissection and "virtual microscopy". I will also be discussing the use of TeleMouse our store forward media rich collaborative software and its use in conjunction with the "Visible Mouse" and "Virtual Microscopy"
Miniaturization: Technological Advances for Physiologic Monitoring of Rodents in Biomedical Research
Tamara Goode, DVM, MS
Merck Research Laboratories
With the increased use of transgenic and knockout models in biomedical research, mice and rats are becoming the animal models of choice in many physiological and pharmacological experiments. However, due to their small size, measurement of in vivo clinical indices and anatomical and physiologic parameters has been difficult in each of these species. We introduce a plan called miniaturization which has allowed us to combine microsurgical techniques with new advances in bioengineering for the development of rodent animal models. The miniaturization plan has enabled us to develop rodent animal models that would traditionally be developed in large animal species. Through miniaturization, we have been able to successfully reduce and refine the number of animals used in biomedical research. The objectives of this seminar are to discuss the conceptual advantages and challenges encountered with the miniaturization plan. In addition, applications using microsurgical techniques, sonomicrometry, magnetic resonance microscopy, telemetry, and echocardiography used in the development of miniaturized animal models for biomedical research will be covered.
Towards remote phenotypic assessment of conscious mice
Tom Hampton
Mouse Specifics, Inc.
The paradigm for physiological characterization of mouse models is changing. With tens of millions of mice being developed for gene and drug target validation studies, non-invasive, high-throughput tools and methods could clearly increase the quantity and improve the quality of data provided by mouse models. Especially as strain, gender, and age-dependent variations affect gene function, screening of conscious animals in relatively unperturbed environmental conditions may provide early clues as to the effects of genetic or pharmacological interventions on the animals' health.
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Dissecting signaling pathways through cell specific gene targeting in mice
Lothar Hennighausen
National Insititutes of Health, Laboratory of Biochemistry & Metabolism
Signaling pathways in normal physiology and cancer can be investigated through manipulations of the mouse genome. However, in many cases the deletion of genes from every cell, as achieved with general gene targeting techniques, does not provide useful information for those concerned with adult physiology and cancer progression. For example, embryonic lethality and indirect effects can hinder the investigation. Such drawbacks can now be overcome through targeting genetic manipulations to the cell types under investigation, and at defined time points. In addition, the grafting of gene deletion cells into a wild-type mouse permits the rescue of otherwise embryonic lethality and the investigation of cell autonomy. Our team has established gene targeting techniques aimed at specific cell types in the adult and embryonic mouse. We have used the Cre-loxP recombination system to understand the role of the cell survival and death molecules in diverse settings, such as germ cells, hematopoietic cells and the mammary epithelium. This experimental approach has led to a deep insight into the cell-specific nature of survival and death factors at different developmental stages.
Functional and Structural Professor of Radiology Phenotyping with Magnetic Resonance Microscopy
G. Allen Johnson
Duke University
Magnetic resonance microscopy (MRM) has recently been adapted for phenotyping structure and function in both mouse and rats at spatial resolution nearly a million times that of conventional MRI. We will review methods for cardiac and pulmonary functional studies in live animals. Three dimensional structural images of whole, fixed specimens are now be acquired as part of "The Visible Mouse" project at isotropic spatial resolution of 100 microns through any arbitrary plane. Images of isolated organs are available at spatial resolution down to 10 microns. The entire atlas is being made available via the internet.
Utilization of Fluorescent Proteins in studying Gene Function in Mice
Andras Nagy
Samuel Lunenfeld Research Institute, Mount Sinai Hospital
Tagging specific cells and genes by fluorescent protein expression provides a unique opportunity to follow the dynamics of mammalian development and disease processes. Besides being the easiest reporter to follow, it gives the 'time' dimension to our studies, allowing time-lapse photography of complex processes. Genes encoding for several color variants are available now, which further expand the capabilities of the developing assays. Some of these will be presented and their power will be discussed.
Phenotyping The Mouse Vascular System By Using Cre/loxP and GFP Systems: Gene Expression, Signaling and Morphogenesis
Thomas N. Sato
University of Texas Southwestern Medical Center at Dallas
We have developed novel lines of transgenic mice by using Cre/loxP and GFP systems. These transgenic lines allow us to study gene expression, signal transduction pathways and morphogenesis of the mouse vascular system. I will present the description of these transgenic lines and some of our up-to-date data. These systems will be useful in phenotyping various transgenic and knockout lines that may have abnormalities in the vascular system. I will also discuss the possibility that the same principle of these systems can be applied essentially to the phenotyping of all the organ systems.
New ES cell-based technologies for Mutagenizing the Mouse Genome and Deriving Disease Models in Culture
John Schimenti
The Jackson Laboratory
Mouse mutants will play an increasingly prominant role in the quest to define gene functions in the "post-genomics" era. Accordingly, large-scale mouse mutagenesis has gained in popularity as a method to derive models of disease. However, there remain considerable technical, logistical and financial drawbacks to this otherwise powerful approach. We have developed methodologies for efficiently mutagenizing mouse embryonic stem (ES) cells for the production of mouse mutants. This will provide a powerful means to generate models of certain diseases by identifying phenotypic mutations in cell culture.
Automated home-cage Monitoring for Detection of Novel Mouse Mutants
Kevin Seburn
The Jackson Laboratory
The Jackson Laboratory has recently launched a large-scale mutagenesis program to systematically collect novel neurological mouse mutants. Beyond the logistics associated with screening large numbers of mice the greatest challenge lies in the need to detect individual deviant mice from a large population. Currently available behavioral and physiological paradigms are typically only used to reveal group differences. We have developed (in conjunction with Columbus Instruments, Columbus Ohio) a comprehensive cage monitoring system (CCMS) that monitors mice in specialized individual live-in cages. The system allows automated, non-invasive, simultaneous collection of: 1) total, ambulatory and rearing activity, 2) food and water consumption; 3) oxygen and carbon dioxide concentrations. Preliminary investigations using CCMS data collected from known mutant mice demonstrated that a variety of different phenotypes could be detected by application of a multivariate statistical algorithm to these data.
Mice in the Post-Genomics Age: Challenges and Potential for Drug Discovery Efforts
Mary Stevens
Bayer Biotechnology
Now that the first draft of the human genome is finished, the challenge is to not only understand the function of these genes, but also their therapeutic utility. There are many technical approaches that use the mouse as a model system in the search for novel gene function, including increasingly sophisticated transgenics, viral vector mediated over expression and knockouts. Mutant mice generated by these technologies can be evaluated in a plethora of disease models. However, regardless of the mechanism by which mutant mice are generated, appropriate phenotypic analysis remains a major hurdle in assessing gene function. One ever present issue is that of the relatively low throughput of phenotyping. This talk will provide a broad overview of emerging technologies, and will address some of the issues facing laboratories in both academia and pharmaceutical companies.
