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Protease nexin-1 regulates retinal vascular development

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Abstract

We recently identified protease nexin-1 (PN-1) or serpinE2, as a possibly underestimated player in maintaining angiogenic balance. Here, we used the well-characterized postnatal vascular development of newborn mouse retina to further investigate the role and the mechanism of action of PN-1 in physiological angiogenesis. The development of retinal vasculature was analysed by endothelial cell staining with isolectin B4. PN-1-deficient (PN-1−/−) retina displayed increased vascularization in the postnatal period, with elevated capillary thickness and density, compared to their wild-type littermate (WT). Moreover, PN-1−/− retina presented more veins/arteries than WT retina. The kinetics of retinal vasculature development, retinal VEGF expression and overall retinal structure were similar in WT and PN-1−/− mice, but we observed a hyperproliferation of vascular cells in PN-1−/− retina. Expression of PN-1 was analysed by immunoblotting and X-Gal staining of retinas from mice expressing beta-galactosidase under a PN-1 promoter. PN-1 was highly expressed in the first week following birth and then progressively decreased to a low level in adult retina where it localized on the retinal arteries. PCR arrays performed on mouse retinal RNA identified two angiogenesis-related factors, midkine and Smad5, that were overexpressed in PN-1−/− newborn mice and this was confirmed by RT-PCR. Both the higher vascularization and the overexpression of midkine and Smad5 mRNA were also observed in gastrocnemius muscle of PN-1−/− mice, suggesting that PN-1 interferes with these pathways. Together, our results demonstrate that PN-1 strongly limits physiological angiogenesis and suggest that modulation of PN-1 expression could represent a new way to regulate angiogenesis.

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Acknowledgments

The authors thank Dr. Mary Osborne-Pellegrin for critical reading of the manuscript. This work was supported by the Institut National de la Santé et de la Recherche Médicale, University Paris 7 Denis Diderot and HemoFlu ANR-13-BSV3-0011. Sonia Selbonne and Deborah Francois were recipients of fellowships from the Fondation pour la Recherche Médicale (FRM).

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Author notes

  1. William Raoul

    Present address: Université François-Rabelais de Tours, CNRS, GICC UMR 7292, Tours, France

Authors and Affiliations

  1. LVTS, INSERM, U1148, Paris, France

    Sonia Selbonne, Deborah Francois, Yacine Boulaftali, Martine Jandrot-Perrus, Marie-Christine Bouton & Véronique Arocas

  2. Univ Paris Diderot, Sorbonne Paris Cité, Paris, France

    Sonia Selbonne, Deborah Francois, Yacine Boulaftali, Martine Jandrot-Perrus, Marie-Christine Bouton & Véronique Arocas

  3. UMR_S 968, Institut de la Vision, Paris, France

    William Raoul & Florian Sennlaub

  4. Univ Paris 06, UPMC, Paris, France

    William Raoul & Florian Sennlaub

  5. Centre Hospitalier National d’Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, 75012, Paris, France

    William Raoul & Florian Sennlaub

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  1. Sonia Selbonne
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Correspondence to Véronique Arocas.

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Selbonne, S., Francois, D., Raoul, W. et al. Protease nexin-1 regulates retinal vascular development. Cell. Mol. Life Sci. 72, 3999–4011 (2015). https://doi.org/10.1007/s00018-015-1972-5

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  • DOI: https://doi.org/10.1007/s00018-015-1972-5

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