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Implication of VEGF and aquaporin 4 mediating Müller cell swelling to diabetic retinal edema

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Abstract

Purpose

Aquaporin 4 (AQP4), a water channel protein, is known to be expressed in retinal Müller cells. The purpose of this study was to determine the effects of VEGF and AQP4 channels on the volumetric changes in Müller cells.

Methods

Retinas from diabetic rats and a cultured Müller cell line, TR-MUL5, were used in this study. Intravitreal injections of VEGF or PBS were performed on either streptozotocin (STZ)-induced diabetic or normoglycemic rats. Retinal sections were immunostained for anti-glial fibrillary acidic protein (GFAP), anti-AQP4, and anti-VEGF. VEGF protein levels from collected retinas were determined by western blot analysis. Volumetric changes and nitric oxide (NO) levels in cultured Müller cells were determined using flow cytometry (FACS), in the presence or absence of VEGF and TGN-020, a selective AQP4 inhibitor.

Results

In the diabetic rat retina, VEGF immunoreactivity was concentrated in the internal retinal layers, and AQP4 immunoreactivity was higher than controls. The expressions of AQP4 were colocalized with GFAP. Protein levels of VEGF in the hyperglycemic rat retina were significantly higher than controls. FACS analyses showed that exposure to VEGF enlarged Müller cells, while exposure to TGN-020 suppressed the enlargement. Intracellular levels of NO were increased after exposure to VEGF, which was suppressed following the addition of TGN-020.

Conclusion

The observed Müller cell swelling is mediated by VEGF and AQP4.

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Authors and Affiliations

  1. Department of Ophthalmology, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki, Osaka, 569-8686, Japan

    Teruyo Kida, Hidehiro Oku, Taeko Horie, Masanori Fukumoto, Yoshitaka Okuda, Seita Morishita & Tsunehiko Ikeda

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  1. Teruyo Kida
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Correspondence to Teruyo Kida.

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Funding

Grant-in-Aid for Scientific Research (C) (Researcher No. 90610105) from the Japan Society for the Promotion of Science (Tokyo, Japan), and the Osaka Eye Bank Association Fund (Osaka, Japan) provided financial support. The sponsor had no role in the design or conduct of this research.

Conflict of interest

All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

Ethical approval (Animal experiments)

All applicable guidelines of ARVO Statement for the care and use of animals were followed. Our experimental protocols conformed to the Animal Research: Reporting In Vivo Experiments (ARRIVE) guidelines.

All procedures performed in studies involving animals were in accordance with the ethical standards of the Osaka Medical College Committee on the Use and Care of Animals (Approval number: 27114) at which the studies were conducted.

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Kida, T., Oku, H., Horie, T. et al. Implication of VEGF and aquaporin 4 mediating Müller cell swelling to diabetic retinal edema. Graefes Arch Clin Exp Ophthalmol 255, 1149–1157 (2017). https://doi.org/10.1007/s00417-017-3631-z

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