Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice

Fraser, Stuart T.; Hadjantonakis, Anna-Katerina; Sahr, Kenneth E.; Willey, Stephen; Kelly, Olivia G.; Jones, Elizabeth A. V.; Dickinson, Mary E.; Baron, Margaret H.

doi:10.1002/gene.20139

Cited by 82 publications

(94 citation statements)

References 45 publications

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“…We previously reported the development and use of human histone H2B GFP-variant fusions that label active chromatin. These are developmentally neutral, and therefore ideal for tracking cell position, division, and death in vivo in mice (Hadjantonakis and Papaioannou, 2004;Fraser et al, 2005;Plusa et al, 2005). However, to date no system that permits the routine visualization and quantitation of dynamic cell morphology and cell behavior has been established for use in embryonic stem (ES) cells and mice.…”

Section: In Vivo Imaging Of Fluorescent Proteinsmentioning

confidence: 99%

In vivo imaging and differential localization of lipid‐modified GFP‐variant fusions in embryonic stem cells and mice

Rhee

Pirity

Lackan

et al. 2006

Genesis

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155

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SummaryThe visualization of live cell behaviors operating in situ combined with the power of mouse genetics represents a major step toward understanding the mechanisms regulating embryonic development, homeostasis, and disease progression in mammals. The availability of genetically encoded fluorescent protein reporters, combined with improved optical imaging modalities, have led to advances in our ability to examine cells in vivo. We developed a series of lipid-modified fluorescent protein fusions that are targeted to and label the secretory pathway and the plasma membrane, and that are amenable for use in mice. Here we report the generation of two strains of mice, each expressing a spectrally distinct lipid-modified GFP-variant fluorescent protein fusion. The CAG::GFP-GPI strain exhibited widespread expression of a glycosylphosphatidylinositol-tagged green fluorescent protein (GFP) fusion, while the CAG::myr-Venus strain exhibited widespread expression of a myristoyl-Venus yellow fluorescent protein fusion. Imaging of live transgenic embryonic stem (ES) cells, either live or fixed embryos and postnatal tissues demonstrated that glycosylphosphatidyl inositol-and myristoyl-tagged GFP-variant fusion proteins are targeted to and serve as markers of the plasma membrane. Moreover, our data suggest that these two lipid-modified protein fusions are dynamically targeted both to overlapping as well as distinct lipid-enriched compartments within cells. These transgenic strains not only represent high-contrast reporters of cell morphology and plasma membrane dynamics, but also may be used as in vivo sensors of lipid localization. Furthermore, combining these reporters with the study of mouse mutants will be a step forward in understanding the inter-and intracellular behaviors underlying morphogenesis in both normal and mutant contexts.

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Section: In Vivo Imaging Of Fluorescent Proteinsmentioning

confidence: 99%

In vivo imaging and differential localization of lipid‐modified GFP‐variant fusions in embryonic stem cells and mice

Rhee

Pirity

Lackan

et al. 2006

Genesis

Self Cite

148

155

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“…For immunohistochemistry, embryos and individual organs were harvested and processed as described previously (Fraser et al, 2005). They were then transferred to Tissue-Tek OCT embedding medium (Sakura Finetek, Torrance, CA) for 3 hr, placed into embedding moulds, and snap-frozen in isopentane chilled in a liquid nitrogen bath.…”

Section: In Situ Hybridization and Immunochemistrymentioning

confidence: 99%

Tg(Afp‐GFP) expression marks primitive and definitive endoderm lineages during mouse development

Kwon

Fraser

Eakin

et al. 2006

Developmental Dynamics

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100

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“…However, this approach does not elucidate dynamic interactions between endothelial cells and other cells of developing organs. Advances in time-lapse imaging have improved understanding of vasculogenesis, flow-induced vascular remodeling, the genetic programming of angiogenesis, and many other facets of vascular development in the mouse and chick yolk sac and during establishment of the body plan in zebrafish (8)(9)(10)(11)(12)(13)(14)(15)(16)(17). However, culturing internal organs throughout critical and prolonged periods of development has been difficult.…”

mentioning

confidence: 99%

Four-dimensional analysis of vascularization during primary development of an organ, the gonad

Coveney

Cool

Oliver

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

150

165

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Time-lapse microscopy has advanced our understanding of yolk sac and early embryonic vascularization. However, it has been difficult to assess endothelial interactions during epithelial morphogenesis of internal organs. To address this issue we have developed the first time-lapse system to study vascularization of a mammalian organ in four dimensions. We show that vascularization of XX and XY gonads is a highly dynamic, sexually dimorphic process. The XX gonad recruits vasculature by a typical angiogenic process. In contrast, the XY gonad recruits and patterns vasculature by a novel remodeling mechanism beginning with breakdown of an existing mesonephric vessel. Subsequently, in XY organs individual endothelial cells migrate and reaggregate in the coelomic domain to form the major testicular artery. Migrating endothelial cells respect domain boundaries well before they are morphologically evident, subdividing the gonad into 10 avascular regions where testis cords form. This model of vascular development in an internal organ has a direct impact on the current dogma of vascular integration during organ development and presents important parallels with mechanisms of tumor vascularization.organogenesis ͉ ovary ͉ testis I nsights into vascular development and patterning in internal organs have historically relied on xenograft models and static analysis (1-7). However, this approach does not elucidate dynamic interactions between endothelial cells and other cells of developing organs. Advances in time-lapse imaging have improved understanding of vasculogenesis, flow-induced vascular remodeling, the genetic programming of angiogenesis, and many other facets of vascular development in the mouse and chick yolk sac and during establishment of the body plan in zebrafish (8-17). However, culturing internal organs throughout critical and prolonged periods of development has been difficult. The technical challenges of organ culture, compounded by simultaneous live imaging, is a major hurdle in understanding how endothelial cells remodel and integrate in an internal organ undergoing morphogenesis.For more than a decade the urogenital ridge (UGR) explant model has provided a unique system to analyze morphogenesis of an internal organ (18,19). The fact that the UGR can be successfully explanted to culture at the critical sex-determination stage [11.5 days postcoitum (dpc)], using conditions that maintain the normal morphological structure of the gonad, has been the driving force behind its broad adoption in the sexdetermination field. However, with respect to internal organ vascularization, the UGR has received little attention as a model system. In the UGR explant model the gonad retains contact with the mesonephros, the source of the endothelium, and vascularization ex vivo occurs similarly to vascularization in vivo (20). During the period of vascularization, development of the testis and ovary diverge morphologically. Whereas the XX gonad shows few morphological changes during this period, the XY gonad undergoes dramatic epi...

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Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice

Cited by 82 publications

References 45 publications

In vivo imaging and differential localization of lipid‐modified GFP‐variant fusions in embryonic stem cells and mice

In vivo imaging and differential localization of lipid‐modified GFP‐variant fusions in embryonic stem cells and mice

Tg(Afp‐GFP) expression marks primitive and definitive endoderm lineages during mouse development

Four-dimensional analysis of vascularization during primary development of an organ, the gonad

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