In Situ Molecular Analysis

原位分子分析

• 批准号: 8246483
• 负责人: DEBORAH J VEIS
• 金额: $ 18.05万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8246483
• 关键词: Adult; Animal Model; Antibodies; Arthritis; Biology; Bone Tissue; Cartilage; Collaborations; Complement; Consultations; Data; Disease; Extracellular Matrix; Fracture Healing; Frozen Sections; Healed; Histology; Immunofluorescence Immunologic; Immunohistochemistry; In Situ; In Situ Hybridization; Injury; Joints; Knowledge; Ligaments; Limb Development; Medicine; Meniscus structure of joint; Messenger RNA; Metastatic Neoplasm to the Bone; Methods; Microscopic; Microscopy; Molecular; Molecular Analysis; Morphology; Mus; Muscle; Musculoskeletal; Musculoskeletal Diseases; Neoplasm Metastasis; Osteoporosis; Phenotype; Plastics; Proteins; Reagent; Research; Research Personnel; Science; Scientist; Services; Staging; Staining method; Stains; Techniques; Tendon structure; Tissue Stains; Tissues; Training; Training and Education; base; bone; cost effective; healing; human disease; human tissue; member; mineralization; molecular phenotype; mouse model; musculoskeletal injury; protein function; response to injury; skeletal; skeletal tissue; tissue preparation; tissue processing

Form and function are intimately interrelated. However, the histological analyses of our target tissues (bone, cartilage, tendon, ligament, muscle) provide specialized challenges not often encountered when conducting these techniques on non-skeletal tissues. For example, due to the extensive extracellular matrix, these tissues can be very difficult to prepare and section, and bone presents a particular challenge due to mineralization. A specialized knowledge of the stages of development, and limb and joint morphology is necessary for the analysis of skeletal tissues. In addition to morphology, the localization of proteins and mRNA in mouse models of musculoskeletal diseases is a vital component in the analysis of the molecular mechanism of protein function, the response to injury, and disease. The overall objective of the In situ Molecular Analysis Core is to provide histological services for the identification and analysis of molecular phenotypes of our target tissues, bone, cartilage, muscle, tendon and ligament, in developing and adult mice. The services will include preparation of tissues and tissue sections; a reagent bank for histology, immunohistochemistry and in situ hybridization; and training in the techniques of in situ hybridization to mRNA, immunohistochemistry, deconvolution microscopy, and histomorphometry. Reagents for the analysis of mouse, and in some cases human, tissues will be available. We expect that the findings from mouse models will be extended to human disease and injury. The Director/Co-Directors and collaborating investigators will provide consultation and advice on analysis and interpretation of the target tissue phenotypes. Deconvolution microscopic analysis and bone histomorphometry will provide the opportunity to utilize sophisticated techniques and quantitative analysis not currently available to all investigators. The strength of this Core lies in the specific techniques developed for adult mice, sectioning delicate early healing tissue, and properly aligning blocks for sectioning of ligament and fracture healing, as well as the expertise in tissue staining by a wide variety of histochemical and molecular techniques. All aspects of the core provide collaboration, training, and education, which will strengthen ties among the research base and enhance the quality of the science performed. Thus, not only will this Core be cost-effective, it will allow scientists to explore skeletal phenotypes that they could not in any other way investigate. This core will complement the specific functional analyses proposed in Core B and the mouse models of Core D.

形式和功能是密切相关的。然而，我们的靶组织的组织学分析 (bone软骨、肌腱、韧带、肌肉）提供了在以下情况下不经常遇到的专门挑战： 在非骨骼组织上进行这些技术。例如，由于广泛的细胞外 基质，这些组织可能非常难以制备和切片，并且骨呈现出特别的挑战 由于矿化。对发育阶段、肢体和关节的专门知识 形态学对于骨骼组织的分析是必要的。除了形态学， 蛋白质和mRNA在小鼠模型的肌肉骨骼疾病是一个重要组成部分，在分析 蛋白质功能的分子机制，对损伤和疾病的反应。总体目标 的原位分子分析核心是提供组织学服务的鉴定和 我们的目标组织，骨，软骨，肌肉，肌腱和韧带的分子表型分析， 发育和成年小鼠。这些服务将包括准备组织和组织切片; a 组织学、免疫组织化学和原位杂交试剂库;技术培训 mRNA原位杂交，免疫组织化学，去卷积显微镜， 组织形态计量学用于分析小鼠组织和在某些情况下人组织的试剂将在本文中公开。 available.我们希望小鼠模型的发现将扩展到人类疾病和损伤。 主任/共同主任和合作调查员将提供分析咨询和建议 和靶组织表型的解释。去卷积显微分析与骨 组织形态计量学将提供利用复杂技术和定量分析的机会 目前还没有提供给所有研究人员。这个核心的力量在于具体的技术 为成年小鼠开发，切片精细的早期愈合组织，并适当对齐块， 韧带和骨折愈合的切片，以及各种组织染色的专业知识， 组织化学和分子技术。核心的所有方面都提供协作、培训和 教育，加强研究基地之间的联系，提高科学质量 执行。因此，这一核心不仅具有成本效益，还将使科学家能够探索骨骼 他们无法以其他方式研究的表型。这一核心将补充具体的 核心B和核心D的小鼠模型中提出的功能分析。