CORE--Tissue Culture

用于转基因和基因敲除动物的世代，但核心也将 在转基因和基因敲除的初步表征研究中支持个别项目 模特们。每个鼠标模型的特征将包括对表达式的仔细分析 转基因模式以及由于以下原因而发生的病理和生理变化 转基因表达。在过去的几年里，圆周派、马克斯博士、马克思和卡斯在 MK24=mink-KVLQT1转基因小鼠的建立及鉴定(1)和 FKBP12.6基因敲除小鼠(2，3)。转基因小鼠的生理特征需要 核心实验室现有的技术专长(4-6人)。过去三年的系统 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PPG提供支助： A.转基因和基因敲除小鼠的制备和维持 B.转基因和基因敲除小鼠的特征，包括-制备和 动物组织标本的分析 C.基线生理测量，包括血流动力学和 超声心动图 D.用于药物测试的转基因和基因敲除小鼠的生理测量。 心肌梗死和必要时的主动脉环扎术 F.血压测量 G.细胞培养系的维护 动物模型和组织培养核心的一个主要目标是开发和测试动物模型 提供研究离子通道中变构相互作用的模型。此核心将填充一个 哥伦比亚大学PPG通过促进遗传模型在 动物使用最先进的技术进行生理测量(例如，心电记录， EPS、超声心动图和血流动力学测量)。最后，我们将维护所有单元格 建立稳定的转基因细胞系，为待研究的各种结构物建立稳定的转基因细胞系。马厩 马克思博士已经建立了野生型表达BK通道的细胞系。 表1.每个项目使用的鼠标型号 项目1-卡林项目2-标记项目3-卡斯项目4-马克思 **(组织培养)**(组织培养)**(组织培养) *豚鼠 J32AR*X X X RyR2-S2809A*X X MK24*X FKBP12.6KO*X KCNQ1-L2M 阶梯连接X 型号* 未来的模式 RyR2-ARVD2 X RyR2-CPVT X BKA淘汰赛X RyR2-S2809D*X 32AR淘汰赛X PHS 398/2590(05/01版)第273页延续格式第‘

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'