Imaging in Developmental Toxicology

发育毒理学成像

• 批准号: 7748137
• 负责人: DEBASHISH ROY
• 金额: $ 19.66万
• 依托单位: BIOINVISION, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7748137
• 关键词: Affect; Analysis of Variance; Anatomy; Animals; Benzoates; Benzyl Alcohols; Biological Markers; Blinded; Blood Vessels; CYP1A1 gene; Cardiac; Cardiovascular system; Cells; Color; Computer software; Congenital Heart Defects; Control Groups; Coronary; Data; Data Set; Defect; Development; Dissection; Dose; Embryo; Embryonic Development; Endothelial Cells; Engineering; Ensure; Exposure to; Face; Fetus; Fluorescence; Fluorescent in Situ Hybridization; Freezing; Gene Expression; Genes; Genetically Engineered Mouse; Glossary; Gray unit of radiation dose; Green Fluorescent Proteins; Hypoxia Inducible Factor; Image; Imagery; Imaging technology; Intestines; Kidney; Laboratory Research; Language; Lighting; Link; Magnetic Resonance Imaging; Mammals; Manuals; Marketing; Maternal Exposure; Methods; Microscopy; Mining; Molecular; Molecular Probes; Morphology; Mus; Neurons; Optics; Oral; Pharmaceutical Preparations; Phase; Polyvinyls; Positioning Attribute; Process; Psyche structure; Pyrrolidinones; Relative (related person); Reporter; Reporter Genes; Reporting; Reproducibility; Research; Research Contracts; Research Personnel; Resolution; Respiratory System; Robot; Robotics; Scientist; Slice; Slide; Smooth Muscle; Smooth Muscle Myocytes; Software Tools; Staging; Staining method; Stains; Stress; Structure; System; Systems Analysis; Technology; Temperature; Testing; Tetrachlorodibenzodioxin; Three-Dimensional Image; Time; Tissues; Toxic effect; Toxicant exposure; Toxicology; Toxin; Transgenic Mice; Transgenic Model; Transgenic Organisms; Universities; Up-Regulation; Variant; Vascular Endothelial Growth Factors; Visualization software; X-Ray Computed Tomography; cryostat; data structure; developmental toxicology; dibenzo(1,4)dioxin; digital; egg; embryo tissue; embryo/fetus; enhanced green fluorescent protein; fetal; fluorescence imaging; fluorescence microscope; gamma Actin; image visualization; improved; instrumentation; interest; malformation; public health relevance; respiratory; response; software development; tissue preparation; tomography; tool; toxicant; urinary; virtual

DESCRIPTION (provided by applicant): Cryo-imaging, as developed by BioInVision, Inc., and fluorescent reporter gene transgenic mice will be used to determine even subtle or rare developmental abnormalities resulting from toxic exposures. By alternately sec- tioning and imaging, the cryo-imaging system will acquire 3D, high-resolution anatomical color and cellular fluorescence images. The cryo-imaging system consists of a large stage, motorized cryostat with special fea- tures for imaging; a modified, bright-field/fluorescence microscope; and a robotic xyz imaging system posi- tioner, all of which are fully automated by a control system. To achieve high throughput, we will modify the sys- tem to acquire images from arrays of as many as 8-50 embryos at a time. There are a very large number of existing fluorescent-reporter-gene transgenic mice which highlight various tissues (smooth muscle cells, endo- thelial cells, neurons, etc.) and emerging transgenic models which fluorescently report upregulation of gene response to toxic stress. With cryo-imaging, we will obtain micron-scale resolution, color and fluorescent vol- umes which will show subtle changes in these engineered mice as a result of toxic exposure. We will develop 3D image visualization/analysis tools allowing us to probe virtual embryos for subtle changes in development. In Phase I, we will demonstrate feasibility using embryos which abundantly expresses EGFP in smooth muscle delineating the fetal vascular, respiratory, and GI systems. Following maternal exposure to TCDD, which is known to affect vascular development, we will analyze fetuses both by conventional manual dissection and by cryo-imaging, and determine the ability of cryo-imaging to detect both gross and subtle variations. Because data is digital, there will be opportunities for computational comparisons of embryos. If this general approach is successful, we will use batteries of fluorescent reporter mice and cryo-imaging to perform toxicology studies providing improved sensitivity and mechanistic information. PUBLIC HEALTH RELEVANCE: We will utilize a new imaging technology and genetically engineered mice to create a very sensitive method for assessing effects of toxins on developing embryos. This research will help drug companies and environmental scientists screen for subtle, rare toxicity effects and to determine general mechanisms of action.

描述（由申请人提供）：由 BioInVision, Inc. 开发的冷冻成像和荧光报告基因转基因小鼠将用于确定因有毒物质暴露而导致的细微或罕见的发育异常。通过交替切片和成像，冷冻成像系统将获取 3D、高分辨率解剖彩色和细胞荧光图像。冷冻成像系统由一个大型电动低温恒温器组成，具有特殊的成像功能；改进的明场/荧光显微镜；以及机器人 xyz 成像系统定位器，所有这些都由控制系统完全自动化。为了实现高通量，我们将修改系统以一次从多达 8-50 个胚胎的阵列中获取图像。现有的荧光报告基因转基因小鼠数量非常多，它们突出显示各种组织（平滑肌细胞、内皮细胞、神经元等），而新兴的转基因模型则荧光报告对毒性应激的基因反应上调。通过冷冻成像，我们将获得微米级的分辨率、颜色和荧光量，这些将显示这些工程小鼠因接触有毒物质而发生的微妙变化。我们将开发 3D 图像可视化/分析工具，使我们能够探测虚拟胚胎发育中的细微变化。在第一阶段，我们将证明使用在描绘胎儿血管、呼吸和胃肠系统的平滑肌中大量表达 EGFP 的胚胎的可行性。母亲接触 TCDD（已知会影响血管发育）后，我们将通过传统手工解剖和冷冻成像来分析胎儿，并确定冷冻成像检测总体和细微变化的能力。由于数据是数字化的，因此将有机会对胚胎进行计算比较。如果这种通用方法成功，我们将使用荧光报告小鼠组和冷冻成像来进行毒理学研究，从而提供更高的灵敏度和机制信息。公共健康相关性：我们将利用新的成像技术和基因工程小鼠来创建一种非常敏感的方法来评估毒素对发育中胚胎的影响。这项研究将帮助制药公司和环境科学家筛选微妙、罕见的毒性效应并确定一般作用机制。