Quantification of oxygen-induced retinopathy in the mouse: a model of vessel loss, vessel regrowth and pathological angiogenesis

Connor, Kip M.; Krah, Nathan M.; Dennison, R. J.; Aderman, Christopher M.; Chen, Jing; Guerin, Karen I.; Sapieha, Przemysław; Stahl, Andreas; Willett, Keirnan; Smith, Lois E.H.

doi:10.1038/nprot.2009.187

Cited by 608 publications

(709 citation statements)

References 26 publications

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“…OIR mouse model, 75% oxygen from 7th post‐natal day to 12th post‐natal day (P7–P12) to induce vaso‐obliteration and room air until P17 to attain maximal preretinal neovascularization was induced as previously described 18. In contrast with control normal mouse, where a paucity of microglia accumulation within the perivascular niche was observed, our data revealed significantly higher numbers of IBA + microglia localized in close proximity to endothelial sprouting pathological tufts during maximal neovascularization at P17 in the retina (Fig.…”

Section: Resultsmentioning

confidence: 99%

Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis

Yin

et al. 2017

J Cellular Molecular Medi

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Retinal microglia cells contribute to vascular angiogenesis and vasculopathy induced by relative hypoxia. However, its concrete molecular mechanisms in shaping retinal angiogenesis have not been elucidated. Basigin, being involved in tumour neovasculogenesis, is explored to exert positive effects on retinal angiogenesis induced by microglia. Therefore, we set out to investigate the expression of basigin using a well‐characterized mouse model of oxygen‐induced retinopathy, which recapitulated hypoxia‐induced aberrant neovessel growth. Our results elucidate that basigin is overexpressed in microglia, which accumulating in retinal angiogenic sprouts. In vitro, conditioned media from microglia BV2 under hypoxia treatment increase migration and tube formation of retinal capillary endothelia cells, compared with media from normoxic condition. The angiogenic capacity of BV2 is inhibited after basigin knockdown by small interfering RNAs. A new molecular mechanism for high angiogenic capacity, whereby microglia cells release basigin via up‐regulation of PI3K‐AKT and IGF‐1 pathway to induce angiogenesis is unveiled. Collectively, our results demonstrate that basigin from hypoxic microglia plays a pivotal pro‐angiogenic role, providing new insights into microglia‐promoting retinal angiogenesis.

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Section: Resultsmentioning

confidence: 99%

Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis

Yin

et al. 2017

J Cellular Molecular Medi

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“…The vascular area was quantified as a percentage of vascularized area compared with total retinal area. Whole-mount retinal images were obtained at 310 magnification using the mosaic function; merge images were imported into Adobe Photoshop for further analysis followed by the established protocol (33). For each group, six retinas were analyzed.…”

Section: Quantification Of Retinal Vascular Developmentmentioning

confidence: 99%

RORα modulates semaphorin 3E transcription and neurovascular interaction in pathological retinal angiogenesis

et al. 2017

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Pathological proliferation of retinal blood vessels commonly causes vision impairment in proliferative retinopathies, including retinopathy of prematurity. Dysregulated crosstalk between the vasculature and retinal neurons is increasingly recognized as a major factor contributing to the pathogenesis of vascular diseases. Class 3 semaphorins (SEMA3s), a group of neuron-secreted axonal and vascular guidance factors, suppress pathological vascular growth in retinopathy. However, the upstream transcriptional regulators that mediate the function of SEMA3s in vascular growth are poorly understood. Here we showed that retinoic acid receptor-related orphan receptor a (RORa), a nuclear receptor and transcription factor, is a novel transcriptional regulator of SEMA3E-mediated neurovascular coupling in a mouse model of oxygen-induced proliferative retinopathy. We found that genetic deficiency of RORa substantially induced Sema3e expression in retinopathy. Both RORa and SEMA3E were expressed in retinal ganglion cells. RORa directly bound to a specific ROR response element on the promoter of Sema3e and negatively regulated Sema3e promoter-driven luciferase expression. Suppression of Sema3e using adeno-associated virus 2 carrying short hairpin RNA targeting Sema3e promoted disoriented pathological neovascularization and partially abolished the inhibitory vascular effects of RORa deficiency in retinopathy. Our findings suggest that RORa is a novel transcriptional regulator of SEMA3E-mediated neurovascular coupling in pathological retinal angiogenesis.-Sun, Y., Liu, C.-H., Wang, Z., Meng, S. S., Burnim, S. B., SanGiovanni, J. P., Kamenecka, T. M., Solt, L. A., Chen, J. RORa modulates semaphorin 3E transcription and neurovascular interaction in pathological retinal angiogenesis. FASEB J. 31, 4492-4502 (2017). www.fasebj.org

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“…Image analysis was performed using a Carl Zeiss Imager Z1 and the imageprocessing software: AxioVision (Carl Zeiss), Photoshop CS (Adobe). Quantification of neovascularization was performed as previously described [13].…”

Section: Zebrafish Lines Morpholinos and Vessel Quantificationmentioning

confidence: 99%

Brag2 differentially regulates β1- and β3-integrin-dependent adhesion in endothelial cells and is involved in developmental and pathological angiogenesis

et al. 2014

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b1-Integrins are essential for angiogenesis. The mechanisms regulating integrin function in endothelial cells (EC) and their contribution to angiogenesis remain elusive. Brag2 is a guanine nucleotide exchange factor for the small Arf-GTPases Arf5 and Arf6. The role of Brag2 in EC and angiogenesis and the underlying molecular mechanisms remain unclear. siRNA-mediated Brag2-silencing reduced EC angiogenic sprouting and migration. Brag2-siRNA transfection differentially affected a5b1-and aVb3-integrin function: specifically, Brag2-silencing increased focal/fibrillar adhesions and adhesion on b1-integrin ligands (fibronectin and collagen), while reducing the adhesion on the aVb3-integrin ligand, vitronectin. Consistent with these results, Brag2-silencing enhanced surface expression of a5b1-integrin, while reducing surface expression of aVb3-integrin. Mechanistically, Brag2-mediated aVb3-integrin-recycling and b1-integrin endocytosis and specifically of the active/matrix-bound a5b1-integrin present in fibrillar/focal adhesions (FA), suggesting that Brag2 contributes to the disassembly of FA via b1-integrin endocytosis. Arf5 and Arf6 are promoting downstream of Brag2 angiogenic sprouting, b1-integrin endocytosis and the regulation of FA. In vivo silencing of the Brag2-orthologues in zebrafish embryos using morpholinos perturbed vascular development. Furthermore, in vivo intravitreal injection of plasmids containing Brag2-shRNA reduced pathological ischemia-induced retinal and choroidal neovascularization. These data reveal that Brag2 is essential for developmental and pathological angiogenesis by promoting EC sprouting through regulation of adhesion by mediating b1-integrin internalization and link for the first time the process of b1-integrin endocytosis with angiogenesis.

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Quantification of oxygen-induced retinopathy in the mouse: a model of vessel loss, vessel regrowth and pathological angiogenesis

Cited by 608 publications

References 26 publications

Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis

Up‐regulated basigin‐2 in microglia induced by hypoxia promotes retinal angiogenesis

RORα modulates semaphorin 3E transcription and neurovascular interaction in pathological retinal angiogenesis

Brag2 differentially regulates β1- and β3-integrin-dependent adhesion in endothelial cells and is involved in developmental and pathological angiogenesis

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