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A transgenic quail model that enables dynamic imaging of amniote embryogenesis.

ABSTRACT: Embryogenesis is the coordinated assembly of tissues during morphogenesis through changes in individual cell behaviors and collective cell movements. Dynamic imaging, combined with quantitative analysis, is ideal for investigating fundamental questions in developmental biology involving cellular differentiation, growth control and morphogenesis. However, a reliable amniote model system that is amenable to the rigors of extended, high-resolution imaging and cell tracking has been lacking. To address this shortcoming, we produced a novel transgenic quail that ubiquitously expresses nuclear localized monomer cherry fluorescent protein (chFP). We characterize the expression pattern of chFP and provide concrete examples of how Tg(PGK1:H2B-chFP) quail can be used to dynamically image and analyze key morphogenetic events during embryonic stages X to 11.

SUBMITTER: Huss D

PROVIDER: S-EPMC4550965 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature

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A transgenic quail model that enables dynamic imaging of amniote embryogenesis.

Huss David D Benazeraf Bertrand B Wallingford Allison A Filla Michael M Yang Jennifer J Fraser Scott E SE Lansford Rusty R

Development (Cambridge, England) 20150724 16

Embryogenesis is the coordinated assembly of tissues during morphogenesis through changes in individual cell behaviors and collective cell movements. Dynamic imaging, combined with quantitative analysis, is ideal for investigating fundamental questions in developmental biology involving cellular differentiation, growth control and morphogenesis. However, a reliable amniote model system that is amenable to the rigors of extended, high-resolution imaging and cell tracking has been lacking. To addr ...[more]

PMID: 26209648

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Project description:BackgroundOne of the least understood and most central questions confronting biologists is how initially simple clusters or sheet-like cell collectives can assemble into highly complex three-dimensional functional tissues and organs. Due to the limits of oxygen diffusion, blood vessels are an essential and ubiquitous presence in all amniote tissues and organs. Vasculogenesis, the de novo self-assembly of endothelial cell (EC) precursors into endothelial tubes, is the first step in blood vessel formation. Static imaging and in vitro models are wholly inadequate to capture many aspects of vascular pattern formation in vivo, because vasculogenesis involves dynamic changes of the endothelial cells and of the forming blood vessels, in an embryo that is changing size and shape.Methodology/principal findingsWe have generated Tie1 transgenic quail lines Tg(tie1:H2B-eYFP) that express H2B-eYFP in all of their endothelial cells which permit investigations into early embryonic vascular morphogenesis with unprecedented clarity and insight. By combining the power of molecular genetics with the elegance of dynamic imaging, we follow the precise patterning of endothelial cells in space and time. We show that during vasculogenesis within the vascular plexus, ECs move independently to form the rudiments of blood vessels, all while collectively moving with gastrulating tissues that flow toward the embryo midline. The aortae are a composite of somatic derived ECs forming its dorsal regions and the splanchnic derived ECs forming its ventral region. The ECs in the dorsal regions of the forming aortae exhibit variable mediolateral motions as they move rostrally; those in more ventral regions show significant lateral-to-medial movement as they course rostrally.Conclusions/significanceThe present results offer a powerful approach to the major challenge of studying the relative role(s) of the mechanical, molecular, and cellular mechanisms of vascular development. In past studies, the advantages of the molecular genetic tools available in mouse were counterbalanced by the limited experimental accessibility needed for imaging and perturbation studies. Avian embryos provide the needed accessibility, but few genetic resources. The creation of transgenic quail with labeled endothelia builds upon the important roles that avian embryos have played in previous studies of vascular development.

Project description:BackgroundThe MRI Breast Imaging-Reporting and Data System (BI-RADS) lexicon recommends that a breast MRI protocol contain T2 -weighted and dynamic contrast-enhanced (DCE) MRI sequences. The addition of diffusion-weighted imaging (DWI) significantly improves diagnostic accuracy. This study aims to clarify which descriptors from DCE-MRI, DWI, and T2 -weighted imaging are most strongly associated with a breast cancer diagnosis.Purpose/hypothesisTo develop a multiparametric MRI (mpMRI) model for breast cancer diagnosis incorporating American College of Radiology (ACR) BI-RADS recommended descriptors for breast MRI with DCE, T2 -weighted imaging, and DWI with apparent diffusion coefficient (ADC) mapping.Study typeRetrospective.SubjectsIn all, 188 patients (mean 51.6 years) with 210 breast tumors (136 malignant and 74 benign) who underwent mpMRI from December 2010 to September 2014.Field strength/sequenceIR inversion recovert DCE-MRI dynamic contrast-enhanced magnetic resonance imaging VIBE Volume-Interpolated-Breathhold-Examination FLASH turbo fast-low-angle-shot TWIST Time-resolved angiography with stochastic Trajectories.AssessmentTwo radiologists in consensus and another radiologist independently evaluated the mpMRI data. Characteristics for mass (n = 182) and nonmass (n = 28) lesions were recorded on DCE and T2 -weighted imaging according to BI-RADS, as well as DWI descriptors. Two separate models were analyzed, using DCE-MRI BI-RADS descriptors, T2 -weighted imagines, and ADCmean as either a continuous or binary form using a previously published ADC cutoff value of ≤1.25 × 10-3 mm2 /sec for differentiation between benign and malignant lesions. Histopathology was the standard of reference.Statistical testsχ2 test, Fisher's exact test, Kruskal-Wallis test, Pearson correlation coefficient, multivariate logistic regression analysis, Hosmer-Lemeshow test of goodness-of-fit, receiver operating characteristics analysis.ResultsIn Model 1, ADCmean (P = 0.0031), mass margins with DCE (P = 0.0016), and delayed enhancement with DCE (P = 0.0016) were significantly and independently associated with breast cancer diagnosis; Model 2 identified ADCmean (P = 0.0031), mass margins with DCE (P = 0.0012), initial enhancement (P = 0.0422), and delayed enhancement with DCE (P = 0.0065) to be significantly independently associated with breast cancer diagnosis. T2 -weighted imaging variables were not included in the final models. DATA CONCLUSION: mpMRI with DCE-MRI and DWI with ADC mapping enables accurate breast cancer diagnosis. A model using quantitative and qualitative descriptors from DCE-MRI and DWI identifies breast cancer with a high diagnostic accuracy. T2 -weighted imaging does not significantly contribute to breast cancer diagnosis.Level of evidence3 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;49:864-874.

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A transgenic quail model that enables dynamic imaging of amniote embryogenesis.

Publications

A transgenic quail model that enables dynamic imaging of amniote embryogenesis.

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