Serveur d'exploration sur le lymphœdème

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A transgenic Xenopus laevis reporter model to study lymphangiogenesis

Identifieur interne : 005E52 ( Ncbi/Curation ); précédent : 005E51; suivant : 005E53

A transgenic Xenopus laevis reporter model to study lymphangiogenesis

Auteurs : Annelii Ny [Belgique] ; Wouter Vandevelde [Belgique] ; Philipp Hohensinner [Belgique] ; Manu Beerens [Belgique] ; Ilse Geudens [Belgique] ; Antonio Diez-Juan [Belgique, Espagne] ; Katleen Brepoels [Belgique] ; Stéphane Plaisance [Belgique] ; Paul A. Krieg [États-Unis] ; Tobias Langenberg [Belgique] ; Stefan Vinckier [Belgique] ; Aernout Luttun [Belgique] ; Peter Carmeliet [Belgique] ; Mieke Dewerchin [Belgique]

Source :

RBID : PMC:3773334

Abstract

Summary

The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the Xenopus tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent Xenopus reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic Tg(Flk1:eGFP) Xenopus laevis reporter lines expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality, Tg(Flk1:eGFP) reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The Tg(Flk1:eGFP) tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening.


Url:
DOI: 10.1242/bio.20134739
PubMed: 24143274
PubMed Central: 3773334

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PMC:3773334

Le document en format XML

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<name sortKey="Krieg, Paul A" sort="Krieg, Paul A" uniqKey="Krieg P" first="Paul A." last="Krieg">Paul A. Krieg</name>
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<country xml:lang="fr">États-Unis</country>
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<name sortKey="Langenberg, Tobias" sort="Langenberg, Tobias" uniqKey="Langenberg T" first="Tobias" last="Langenberg">Tobias Langenberg</name>
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<name sortKey="Vinckier, Stefan" sort="Vinckier, Stefan" uniqKey="Vinckier S" first="Stefan" last="Vinckier">Stefan Vinckier</name>
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<name sortKey="Luttun, Aernout" sort="Luttun, Aernout" uniqKey="Luttun A" first="Aernout" last="Luttun">Aernout Luttun</name>
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<name sortKey="Dewerchin, Mieke" sort="Dewerchin, Mieke" uniqKey="Dewerchin M" first="Mieke" last="Dewerchin">Mieke Dewerchin</name>
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<title xml:lang="en" level="a" type="main">A transgenic
<italic>Xenopus laevis</italic>
reporter model to study lymphangiogenesis</title>
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<name sortKey="Beerens, Manu" sort="Beerens, Manu" uniqKey="Beerens M" first="Manu" last="Beerens">Manu Beerens</name>
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<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
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<country xml:lang="fr">Belgique</country>
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<name sortKey="Geudens, Ilse" sort="Geudens, Ilse" uniqKey="Geudens I" first="Ilse" last="Geudens">Ilse Geudens</name>
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<country>Belgium</country>
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<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<name sortKey="Diez Juan, Antonio" sort="Diez Juan, Antonio" uniqKey="Diez Juan A" first="Antonio" last="Diez-Juan">Antonio Diez-Juan</name>
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,
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<country>Belgium</country>
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<country xml:lang="fr">Belgique</country>
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<affiliation wicri:level="1">
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,
<country>Belgium</country>
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<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<affiliation wicri:level="1">
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<addr-line>Present address: Instituto de Investigación Sanitaria INCLIVA, 46010 Valencia, Spain</addr-line>
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<country xml:lang="fr">Espagne</country>
<wicri:regionArea>Present address: Instituto de Investigación Sanitaria INCLIVA, 46010 Valencia</wicri:regionArea>
<wicri:noRegion>46010 Valencia</wicri:noRegion>
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<name sortKey="Brepoels, Katleen" sort="Brepoels, Katleen" uniqKey="Brepoels K" first="Katleen" last="Brepoels">Katleen Brepoels</name>
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,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
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<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<addr-line>KU Leuven, 3000 Leuven</addr-line>
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<country>Belgium</country>
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</author>
<author>
<name sortKey="Plaisance, Stephane" sort="Plaisance, Stephane" uniqKey="Plaisance S" first="Stéphane" last="Plaisance">Stéphane Plaisance</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff2">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Krieg, Paul A" sort="Krieg, Paul A" uniqKey="Krieg P" first="Paul A." last="Krieg">Paul A. Krieg</name>
<affiliation wicri:level="1">
<nlm:aff id="aff4">
<institution>Department of Cell Biology and Anatomy, University of Arizona College of Medicine</institution>
,
<addr-line>Tucson, AZ 85724-5044</addr-line>
,
<country>USA</country>
</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Langenberg, Tobias" sort="Langenberg, Tobias" uniqKey="Langenberg T" first="Tobias" last="Langenberg">Tobias Langenberg</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff2">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Vinckier, Stefan" sort="Vinckier, Stefan" uniqKey="Vinckier S" first="Stefan" last="Vinckier">Stefan Vinckier</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff2">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Luttun, Aernout" sort="Luttun, Aernout" uniqKey="Luttun A" first="Aernout" last="Luttun">Aernout Luttun</name>
<affiliation wicri:level="1">
<nlm:aff id="aff3">
<institution>Center for Molecular and Vascular Biology</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Carmeliet, Peter" sort="Carmeliet, Peter" uniqKey="Carmeliet P" first="Peter" last="Carmeliet">Peter Carmeliet</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff2">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Dewerchin, Mieke" sort="Dewerchin, Mieke" uniqKey="Dewerchin M" first="Mieke" last="Dewerchin">Mieke Dewerchin</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>VIB, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff2">
<institution>Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center</institution>
,
<addr-line>KU Leuven, 3000 Leuven</addr-line>
,
<country>Belgium</country>
</nlm:aff>
<country xml:lang="fr">Belgique</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series>
<title level="j">Biology Open</title>
<idno type="eISSN">2046-6390</idno>
<imprint>
<date when="2013">2013</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc>
<textClass></textClass>
</profileDesc>
</teiHeader>
<front>
<div type="abstract" xml:lang="en">
<title>Summary</title>
<p>The importance of the blood- and lymph vessels in the transport of essential fluids, gases, macromolecules and cells in vertebrates warrants optimal insight into the regulatory mechanisms underlying their development. Mouse and zebrafish models of lymphatic development are instrumental for gene discovery and gene characterization but are challenging for certain aspects, e.g. no direct accessibility of embryonic stages, or non-straightforward visualization of early lymphatic sprouting, respectively. We previously demonstrated that the
<italic>Xenopus</italic>
tadpole is a valuable model to study the processes of lymphatic development. However, a fluorescent
<italic>Xenopus</italic>
reporter directly visualizing the lymph vessels was lacking. Here, we created transgenic
<italic>Tg(Flk1:eGFP) Xenopus laevis</italic>
reporter lines expressing green fluorescent protein (GFP) in blood- and lymph vessels driven by the Flk1 (VEGFR-2) promoter. We also established a high-resolution fluorescent dye labeling technique selectively and persistently visualizing lymphatic endothelial cells, even in conditions of impaired lymph vessel formation or drainage function upon silencing of lymphangiogenic factors. Next, we applied the model to dynamically document blood and lymphatic sprouting and patterning of the initially avascular tadpole fin. Furthermore, quantifiable models of spontaneous or induced lymphatic sprouting into the tadpole fin were developed for dynamic analysis of loss-of-function and gain-of-function phenotypes using pharmacologic or genetic manipulation. Together with angiography and lymphangiography to assess functionality,
<italic>Tg(Flk1:eGFP)</italic>
reporter tadpoles readily allowed detailed lymphatic phenotyping of live tadpoles by fluorescence microscopy. The
<italic>Tg(Flk1:eGFP)</italic>
tadpoles represent a versatile model for functional lymph/angiogenomics and drug screening.</p>
</div>
</front>
<back>
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