Ultrastructure

超微结构

基本信息

批准号：
9068773
负责人：
CLARA FRANZINI-ARMSTRONG
金额：
$ 12.81万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-14 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9068773
关键词：
Affect Age Aging Animal Model Apoptosis Area Behavior Biopsy Calcium Cells Collaborations Confocal Microscopy Coupling Data Dependence Detection Development Electron Microscopy Electrons Embryo Evaluation Extravasation Feedback Fiber Foundations Freeze Fracturing Frequencies Gender Goals Grant Guidelines Heterozygote Homozygote Human Human Pathology Image Individual Induced Mutation Instruction Light Microscope Link Malignant hyperpyrexia due to anesthesia Microtomy Mitochondria Modeling Molecular Mouse Strains Mus Muscle Muscle Fibers Mutate Mutation Organelles Pathology Patients Permeability Phase Proteins Reaction RyR1 Ryanodine Receptor Calcium Release Channel SERCA1 Sampling Sarcoplasmic Reticulum Skeletal Muscle Staging Surface Techniques Testing Triad Acrylic Resin Variant Work base density early onset experience high standard human tissue insight interest morphometry mouse model overexpression protein protein interaction response sex

项目摘要

Core D will perform the qualitative and quantitative histological and ultrastructural checks that are necessary to support all other Projects. The Core will not only supply various techniques of electron, phase contrast and confocal microscopy, but it will also operate at a high standard of quality and offer an expert critical evaluation of the results of experimental and molecular alterations. The uniform and consistent use of high quality images will allow the detection of even subtle alterations in protein-protein interactions and in the response of individual cell organelles that either are at the basis of altered functions or are the long term results of such alterations. It is clear that the overall ultrastructural response of the muscle fiber to mutations affecting excitation-contraction coupling are quite specific and offer considerable insight into causative effects. In the past period we have evidenced a strong fiber type- and gender-dependence of the pathology, that correspindes quite well with similar variations in function. Two general approaches are proposed. One is to define any alterations in the relationships between the major protein components of calcium release units (CRUs, triads in skeletal muscle) within the context of the mutation and the other is to follow the development of pathology (most specifically mitochondrial, myofibrillar and CRUs' alterations) through development and aging and in relation to the known functional effects of the mutation on CRUs' channels.This will be achieved by combining light microscope techniques (phase contrast of fibers whole mounts and confocal imaging of fluorescently immunolabeled fibers) with thin sectioning and freeze-fracture for electron microscopy supplemented by quantitative morphometry techniques. The core aims at defining the primary impact of each mutation on the macromelcular assembly of calcium release units within a short term and the secondary impact on SR, mitochondria and contractile material on the long term.

核心D将执行支持所有其他项目所需的定性和定量组织学和超微结构检查。核心不仅将提供各种电子技术，相位对比度和共聚焦显微镜，而且还将以高质量标准运行，并对实验和分子改变的结果提供专家的批判性评估。高质量图像的统一使用将允许检测蛋白质 - 蛋白质相互作用的细微变化以及在功能改变的基础上的单个细胞细胞器的响应中，或者是这种改变的长期结果。显然，肌肉纤维对影响激发反应耦合的突变的总体超微结构反应非常具体，并提供了相当大的洞察力。在过去的时期，我们已经证明了该病理的强纤维类型和性别依赖性，这与功能的类似变化相关。提出了两种一般方法。 One is to define any alterations in the relationships between the major protein components of calcium release units (CRUs, triads in skeletal muscle) within the context of the mutation and the other is to follow the development of pathology (most specifically mitochondrial, myofibrillar and CRUs' alterations) through development and aging and in relation to the known functional effects of the mutation on CRUs' channels.This will be achieved by combining light显微镜技术（整个纤维的相位对比度和荧光免疫标记的纤维的共聚焦成像），用薄的截面和冻结骨折的电子显微镜，并补充了通过定量形态计量学补充的电子显微镜。核心的目的是在短期内定义每个突变对钙释放单元的大型钙释放单元的主要影响，并长期对SR，线粒体和收缩材料的次要影响。