Use of the mouse aortic ring assay to study angiogenesis

Baker, Marianne; Robinson, Stephen D.; Lechertier, Tanguy; Barber, Paul R.; Tavora, Bernardo; D’Amico, Gabriela; Jones, Dylan T.; Vojnovic, B.; Hodivala‐Dilke, Kairbaan

doi:10.1038/nprot.2011.435

Cited by 410 publications

(426 citation statements)

References 48 publications

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“…Thoracic aortae from 5-7-weekold C57/BL6 mice (The Jackson Laboratory) were surgically isolated, cleaned, and dissected into 0.5-mm rings. Rings were embedded in 1 mg/ml of type I collagen in a 96-well plate as described previously (84). When embedded, the rings were divided into respective groups: vehicle (PBS or DMSO), endorepellin (200 nM), Torin 1 (40 nM), compound C (1 M), and compound C ϩ endorepellin.…”

Section: Methodsmentioning

confidence: 99%

Endorepellin-evoked Autophagy Contributes to Angiostasis

Goyal

Gubbiotti

Chery

et al. 2016

Journal of Biological Chemistry

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Endorepellin, the C-terminal domain of perlecan, is an angiostatic molecule that acts as a potent inducer of autophagy via its interaction with VEGFR2. In this study, we examined the effect of endorepellin on endothelial cells using atomic force microscopy. Soluble endorepellin caused morphological and biophysical changes such as an increase in cell surface roughness and cell height. Surprisingly, these changes were not accompanied by alterations in the endothelial cell elastic modulus. We discovered that endorepellin-induced autophagic flux led to co-localization of mammalian target of rapamycin with LC3-positive autophagosomes. Endorepellin functioned upstream of AMPactivated kinase ␣, as compound C, an inhibitor of AMP-activated kinase ␣, abrogated endorepellin-mediated activation and co-localization of Beclin 1 and LC3, thereby reducing autophagic progression. Functionally, we discovered that both endorepellin and Torin 1, a canonical autophagic inducer, blunted ex vivo angiogenesis. We conclude that autophagy is a novel mechanism by which endorepellin promotes angiostasis independent of nutrient deprivation.

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Section: Methodsmentioning

confidence: 99%

Endorepellin-evoked Autophagy Contributes to Angiostasis

Goyal

Gubbiotti

Chery

et al. 2016

Journal of Biological Chemistry

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“…Aortic rings from VimKO and WT mice were excised and embedded in collagen containing growth factors to induce sprouting (57). The assay was optimized using different growth factor supplements to ensure appropriate conditions for endothelial outgrowth of endothelial-lined tubes (Fig.…”

Section: Deletion Of Vimentin Disrupts Sprouting Angiogenesis and Attmentioning

confidence: 99%

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

Antfolk

Sjöqvist

Cheng

et al. 2017

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

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Notch signaling is a key regulator of angiogenesis, in which sprouting is regulated by an equilibrium between inhibitory Dll4-Notch signaling and promoting Jagged-Notch signaling. Whereas Fringe proteins modify Notch receptors and strengthen their activation by Dll4 ligands, other mechanisms balancing Jagged and Dll4 signaling are yet to be described. The intermediate filament protein vimentin, which has been previously shown to affect vascular integrity and regenerative signaling, is here shown to regulate ligand-specific Notch signaling. Vimentin interacts with Jagged, impedes basal recycling endocytosis of ligands, but is required for efficient receptor ligand transendocytosis and Notch activation upon receptor binding. Analyses of Notch signal activation by using chimeric ligands with swapped intracellular domains (ICDs), demonstrated that the Jagged ICD binds to vimentin and contributes to signaling strength. Vimentin also suppresses expression of Fringe proteins, whereas depletion of vimentin enhances Fringe levels to promote Dll4 signaling. In line with these data, the vasculature in vimentin knockout (VimKO) embryos and placental tissue is underdeveloped with reduced branching. Disrupted angiogenesis in aortic rings from VimKO mice and in endothelial 3D sprouting assays can be rescued by reactivating Notch signaling by recombinant Jagged ligands. Taken together, we reveal a function of vimentin and demonstrate that vimentin regulates Notch ligand signaling activities during angiogenesis.

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“…Aortic ring assay Dissection of the aorta, serum starvation, embedding and feeding of the aortic rings, imaging and quantifying microvessel sprouts was as described previously [25]. A total of 15 rings per mouse were obtained.…”

mentioning

confidence: 99%

“…The microvessel growth from the aorta rings was quantified on days 5, 6, 7 and 8 post embedding by live phase-contrast microscopy. Emerging microvessels were counted and the data was plotted as mean microvessel numbers per ring as described previously [25].…”

mentioning

confidence: 99%

Attenuation of flightless I improves wound healing and enhances angiogenesis in a murine model of type 1 diabetes

et al. 2013

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Aims/hypothesis Skin lesions and ulcerations are severe complications of diabetes that often result in leg amputations. In this study we investigated the function of the cytoskeletal protein flightless I (FLII) in diabetic wound healing. We hypothesised that overexpression of FLII would have a negative effect on diabetic wound closure and modulation of this protein using specific FLII-neutralising antibodies (FnAb) would enhance cellular proliferation, migration and angiogenesis within the diabetic wound.Methods Using a streptozotocin-induced model of diabetes we investigated the effect of altered FLII levels through Flii genetic knockdown, overexpression or treatment with FnAb on wound healing. Diabetic wounds were assessed using histology, immunohistochemistry and biochemical analysis. In vitro and in vivo assays of angiogenesis were used to assess the angiogenic response. Results FLII levels were elevated in the wounds of both diabetic mice and humans. Reduction in the level of FLII improved healing of murine diabetic wounds and promoted a robust pro-angiogenic response with significantly elevated von Willebrand factor (vWF) and vascular endothelial growth factor (VEGF)-positive endothelial cell infiltration. Diabetic mouse wounds treated intradermally with FnAb showed improved healing and a significantly increased rate of reepithelialisation. FnAb improved the angiogenic response through enhanced formation of capillary tubes and functional neovasculature. Reducing the level of FLII led to increased numbers of mature blood vessels, increased recruitment of smooth muscle actin-α-positive cells and improved tight junction formation. Conclusions/interpretation Reducing the level of FLII in a wound may be a potential therapeutic approach for the treatment of diabetic foot ulcers.

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Use of the mouse aortic ring assay to study angiogenesis

Cited by 410 publications

References 48 publications

Endorepellin-evoked Autophagy Contributes to Angiostasis

Endorepellin-evoked Autophagy Contributes to Angiostasis

Selective regulation of Notch ligands during angiogenesis is mediated by vimentin

Attenuation of flightless I improves wound healing and enhances angiogenesis in a murine model of type 1 diabetes

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