CORE--Tissue Culture

用于产生转基因和基因敲除动物，但核心也将 在转基因和基因敲除的初步表征研究中支持各个项目 模型每个小鼠模型的表征将包括对表达的仔细分析。 转基因的模式和病理和生理变化，这是由于 转基因表达PI，Marks博士，马克思和卡斯在过去的几年里合作， MK24= MinK-KVLQT 1转基因小鼠的产生和表征（1），以及 FKBP 12.6敲除小鼠（2，3）。转基因小鼠的生理特性需要 堆芯实验室中存在的技术专门知识（4-6）。在过去三年中， 已经在实验室里建立起来，让我们能够进行这样的研究。' 该核心将在以下领域为方案规划小组提供支助： 是的转基因和基因敲除小鼠的制备和维持 B。转基因和敲除小鼠的表征，包括制备和 动物组织标本分析 C.基线生理测量，包括血液动力学和 超声心动 D.用于药物测试的转基因和基因敲除小鼠的生理学测量。 心肌梗死和主动脉结扎术（必要时） F.血压测量 G.细胞培养系的维持 动物模型和组织培养核心的主要目标是开发和测试动物模型， 提供模型，其中检查变构相互作用的离子通道。这个核心将填补一个 哥伦比亚大学PPG通过促进遗传模型的使用， 使用现有技术进行生理测量（例如，ECG记录， EPS、超声心动图和血液动力学测量）。最后，我们将保持所有细胞 细胞系，并为待研究的各种构建体开发稳定的转染细胞系。稳定的 马克思博士已经开发了用野生型表达BK.channel转染的细胞系。 表1.每个项目使用的小鼠模型 项目1-Karlin项目2-Marks项目3-Kass项目4-马克思 **（组织培养）**（组织培养）**（组织培养） [*]阿斯托里亚酒店 J32AR* X X RyR2-S2809A* X X MK24* X FKBP 12.6KO * X KCNQ1-L2M LAD结扎X 型号 * 未来车型 RyR2-ARVD2 X RyR 2-CPVT X BKa敲除X RyR2-S2809D* X 32AR敲除X PHS 398/2590（05/01修订版）第273页

for the generation of transgenic and knockout animals but the core will also support the individual projects in the initial characterization studies on the transgenic and knockout models. The characterization of each mouse model will include careful analysis of the expression pattern of the transgene and pathological and physiological changes, which occur as a result of transgene expression. The PI, Dr. Marks, Marx and Kass have collaborated over the past few years on the generation and characterization of the MK24=MinK-KVLQTl transgenic mouse (1) and FKBP12.6 knockout mice (2, 3). Physiological characterization of transgenic mice requires the technical expertise that is present in the laboratory of the core (4-6). Over the past three years systems have been set up in the laboratory to allow us to perform such studies. ' The core will provide support for the PPG in the following areas: ¿ A. The preparation and maintenance of transgenic and knockout mice B. Characterization of transgenic and knock-out mice including -preparation and analysis of tissue specimens from animals C. Baseline physiological measurements including hemodynamics and echocardiography D. Physiological measurements on transgenic and knockout mice for drug testing. ¿ Myocardial infarction and aortic banding when necessary F. Blood Pressure Measurements G. Maintenance of Cell CultureLines A major goal of the Animal Models and Tissue Culture Core is to develop and test animal models to provide models in which to examine allosteric interactions in ion channels. This Core will fill an essential and exciting role of the Columbia University PPG by facilitating the use of genetic models in animals using state-of-the-art techniques to make physiological measurements (eg,.ECG recording, EPS, echocardiograms and hemodynamic measurements) in mice. Finally, we will maintain all cell lines and develop stable transfected cell lines for the various constructs to be investigated. A stable cell line transfected with a wild-type expressing BK.channel has already been developed by Dr. Marx. Table 1. Mouse models used by each project Project 1-Karlin Project 2-Marks Project 3-Kass Project 4-Marx **(Tissue Culture) **(Tissue Culture) **(Tissue Culture) ***GuineaPigs J32AR* X X RyR2-S2809A* X X MK24* X FKBP12.6KO* X KCNQ1-L2M LAD Ligation X Model* Future models RyR2-ARVD2 X RyR2-CPVT X BKa knockout X RyR2-S2809D* X 32AR knockout X PHS 398/2590 (Rev. 05/01) Page 273 Continuation Format Page'