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.
Public Health Relevance Statement: Project Narrative 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.
NIH Spending Category: Agent Orange & Dioxin; Bioengineering; Genetics
Project Terms: 2,3,7,8-Tetrachlorodibenzo-p-dioxin; 3D image; AHH; AHRR; ANOVA; Affect; Analysis of Variance; Anatomic; Anatomical Sciences; Anatomy; Animals; Benzoates; Benzyl Alcohols; Blinded; Blood Vessels; Body Tissues; CAT Scan, X-Ray; CAT scan; CP11; CT X Ray; CT scan; CY11; CYP1; CYP1A1; CYP1A1 gene; Cardiac; Cardiac Abnormalities; Cardiac Defects, Congenital; Cardiac Malformation; Cardiac defect; Cardiovascular; Cardiovascular Body System; Cardiovascular system; Cardiovascular system (all sites); Cells; Color; Computed Tomography; Computer Programs; Computer Software Tools; Computer software; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Congenital Heart Defects; Control Groups; Coronary; DISSEC; Data; Data Set; Dataset; Defect; Development; Dibenzo(b,e)(1,4)dioxin, 2,3,7,8-tetrachloro-; Dissection; Dose; Drugs; EGFP protein; EMI scan; Embryo; Embryo Development; Embryogenesis; Embryonic; Embryonic Development; Embryonic Tissue; Endothelial Cells; Engineering; Engineerings; Ensure; Exposure to; FISH Technic; FISH Technique; FISH analysis; Face; Fetus; Fluorescence; Fluorescent in Situ Hybridization; Freezing; GFP; Gene Expression; Genes; Genetically Engineered Mouse; Glossary; Gray; Gray unit of radiation dose; Green Fluorescent Proteins; Heart Abnormalities; Heart Defects, Congenital; Heart Malformation; Hypoxia Inducible Factor; Illumination; Image; Imagery; Images, 3-D; Imaging technology; In Situ Hybridization, Fluorescence; Instrumentation, Other; Intestinal; Intestines; Investigators; Kidney; Laboratory Research; Language; Leiomyocyte; Lighting; Link; MR Imaging; MR Tomography; MRI; Magnetic Resonance Imaging; Magnetic Resonance Imaging Scan; Mammalia; Mammals; Mammals, General; Mammals, Mice; Manuals; Marketing; Maternal Exposure; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; Medication; Methods; Mice; Microscopy; Mining; Minings; Molecular; Molecular Probes; Morphology; Murine; Mus; Muscle, Involuntary; Muscle, Smooth; Myocytes, Smooth Muscle; NMR Imaging; NMR Tomography; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; Neurons; Nuclear Magnetic Resonance Imaging; Optics; Oral; Organ System, Cardiovascular; P1-450; P450-C; P450DX; P450D\X; Pharmaceutic Preparations; Pharmaceutical Preparations; Phase; Phenylcarbinols; Phenylmethanols; Polyvinyls; Position; Positioning Attribute; Process; Psyche structure; Pulmonary Body System; Pulmonary Organ System; Pyrrolidinones; Pyrrolidones; Relative; Relative (related person); Reporter; Reporter Genes; Reporting; Reproducibility; Research; Research Contracts; Research Personnel; Researchers; Resolution; Respiratory System; Respiratory system (all sites); Robot; Robotics; Science of Anatomy; Scientist; Slice; Slide; Smooth Muscle Cells; Smooth Muscle Myocytes; Smooth Muscle Tissue Cell; Smooth muscle (tissue); Software; Software Tools; Staging; Staining method; Stainings; Stains; Stress; Structure; System; System, LOINC Axis 4; Systems Analyses; Systems Analysis; TCDD; Technology; Temperature; Testing; Tetrachlorodibenzodioxin; Three-Dimensional Image; Time; Tissues; Tomodensitometry; Tomography, Xray Computed; Tools, Software; Toxic effect; Toxicant exposure; Toxicities; Toxicology; Toxin; Transgenic Mice; Transgenic Model; Transgenic Organisms; UV laboratory microscope; Ultraviolet Microscopes; Universities; Up-Regulation; Up-Regulation (Physiology); Upregulation; Urinary System, Kidney; VEGFs; Variance Analyses; Variant; Variation; Vascular Endothelial Growth Factors; Vascular, Heart; Vegf; Visualization; Visualization software; X-Ray Computed Tomography; Zeugmatography; anatomy; biomarker; bowel; catscan; circulatory system; computed axial tomography; computer program/software; computerized axial tomography; computerized tomography; cryostat; data structure; develop software; developing computer software; developmental toxicology; dibenzo(1,4)dioxin; dibenzo(b,e)(1,4)dioxin; dibenzo-p-dioxin; digital; drug/agent; egg; embryo tissue; embryo/fetus; enhanced green fluorescent protein; facial; fetal; fluorescence imaging; fluorescence microscope; fluorescence/UV microscope; fluorescent microscope; gamma Actin; heart defect; image visualization; imaging; improved; instrumentation; interest; laboratory fluorescence light microscope; malformation; mental; neuronal; public health relevance; renal; respiratory; respiratory tract; response; software development; tissue preparation; tomography; tool; toxic exposure; toxicant; transgenic; urinary; vascular; virtual
