July 2019
Volume 60, Issue 9
Free
ARVO Annual Meeting Abstract  |   July 2019
TRPM2 gene deficiency promotes the growth of laser-induced choroidal neovascularization in mice.
Author Affiliations & Notes
  • KOSUKE NISHI
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • HIroki Iwanishi
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Yuta Usui
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Takayoshi Sumioka
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Masayasu Miyajima
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Yuka Okada
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Shizuya Saika
    Ophthalmology, Wakayama Medical University, Wakayama, Japan
  • Footnotes
    Commercial Relationships   KOSUKE NISHI, None; HIroki Iwanishi, None; Yuta Usui, None; Takayoshi Sumioka, None; Masayasu Miyajima, None; Yuka Okada, None; Shizuya Saika, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 1258. doi:https://doi.org/
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      KOSUKE NISHI, HIroki Iwanishi, Yuta Usui, Takayoshi Sumioka, Masayasu Miyajima, Yuka Okada, Shizuya Saika; TRPM2 gene deficiency promotes the growth of laser-induced choroidal neovascularization in mice.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1258. doi: https://doi.org/.

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

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Abstract

Purpose : To investigate the effects of gene ablation of transient receptor potential M2 (TRPM2) on experimental argon laser-induced choroidal neovascularization (CNV) in mice.

Methods : (1) Male wild-type (WT) mice (n = 6) of 6 - 8 week-old were used. Expression of TRPM2 and PECAM-1(vascular endothelial cell marker) in retino-choroidal tissue was examined by flat mount immunohistochemistry at 3, 5, and 7 days after argon laser-photocoagulation (spot diameter 80 μm, output 200mW, irradiation time 0.05 sec) in WT mice. (2) WT mice (n = 5) and C57BL/6-backgroud TRPM2-null (KO) mice (n = 5) were used to clear the effects of TRPMs gene knockout. FITC-angiography and observation under Apo Tome. 2 (Zeiss, Germany) in the flat mount samples were performed 7, 14 and 21 days after laser-photocoagulation. The size of CNV was analyzed by computer software WinROOF (Mitani, Japan). (3) Argon laser-photocoagulation (25 shots) was performed in an eye of WT (n = 10) and KO (n = 10) mice. Total RNA was extracted from retino-choroidal tissue at day 1 and 3 post-treatment. Real-time RT-PCR was ran to semi-quantify the expression level of mRNAs of MCP-1, TGFβ-1, MPO, VEGF, F4/80, and IL-6.

Results : (1) TRPM2 was expressed in all timepoints examined after laser irradiation in WT mice. No co-staining with PECAM-1 was observed. (2) CNV was significantly larger in KO mice compared to WT mice at day 14 (p < 0.05). (3) The loss of TRPM2 suppressed mRNA expression of MPO (at day 1), F4/80 and IL-6 (at day 3) in retino-choroidal tissue post laser irradiation (p<0.05).

Conclusions : TRPM2 deficiency promotes the growth of laser-induced CNV, although inflammation and IL-6 expression were suppressed in KO tissue.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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