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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.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effects of gene ablation of transient receptor potential M2 (TRPM2) on experimental argon laser-induced choroidal neovascularization (CNV) in mice.
(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.
(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).
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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