June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Annexin V inhibits pathological retinal angiogenesis but not physiological retinal vascular development
Author Affiliations & Notes
  • Satoshi Morooka
    Ophthalmology and Visual Sciences, Kyoto University, Kyoto, Japan
  • Tomoaki Murakami
    Ophthalmology and Visual Sciences, Kyoto University, Kyoto, Japan
  • Masayuki Nukada
    Ophthalmology and Visual Sciences, Kyoto University, Kyoto, Japan
    Shiga Medical Center for Adults, Shiga, Japan
  • Nagahisa Yoshimura
    Ophthalmology and Visual Sciences, Kyoto University, Kyoto, Japan
  • Footnotes
    Commercial Relationships Satoshi Morooka, None; Tomoaki Murakami, None; Masayuki Nukada, None; Nagahisa Yoshimura, Canon (C), Canon (F), Nidek (C), Topcon (F), PCT/JP2011/073160 (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 5609. doi:
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    • Get Citation

      Satoshi Morooka, Tomoaki Murakami, Masayuki Nukada, Nagahisa Yoshimura; Annexin V inhibits pathological retinal angiogenesis but not physiological retinal vascular development. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5609.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Although Annexin V is expressed in perivascular cells, its effects on retinal angiogenesis remain ill-defined. Here we investigated how Annexin V regulates retinal angiogenesis.

Methods: Annexin V or neutralizing antibodies against Annexin V was intravitreally injected to retinopathy of prematurity (ROP) mice (C57BL6/J) on P12, P14, and P16, and eyes were harvested on P17. They were intraperitoneally administrated to neonatal mice on P2, and eyes were harvested on P4. To evaluate the affinity of Annexin V to the retinal vasculature, 1 hour after Cy3-conjugated Annexin V were intravitreally administrated on P17 in ROP mice or P4 in neonatal mice, eyes were fixed with 4% paraformaldehyde and immunostained with isolectin B4. To evaluate its toxicity, human umbilical venous endothelial cells (HUVECs) were cultured and treated with Annexin V, followed by LIVE/DEAD assay.

Results: The areas of neovascular tufts were significantly decreased in retinas treated with Annexin V (p<0.05), whereas the tufts were increased in retinas administrated with neutralizing antibody against Annexin V, with statistical significance (p<0.01). Interestingly, Cy3-conjugated Annexin V was attached specifically to smaller neovascular tufts, but not intraretinal vascular beds. However, the radius of vascularized area of the retinas treated with Annexin V or the neutralizing antibodies was not different from the control. Additionally, Cy3-conjugated Annexin V were not adherent to retinal vasculature including tip cells and stalk cells. In vitro experiments demonstrated that annexin V did not have influences on the survival or cell death in HUVECs.

Conclusions: These data suggest that Annexin V are adherent to smaller neovascular tufts specifically, and inhibits pathological, but not physiological angiogenesis.

Keywords: 700 retinal neovascularization  
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