Involvement of GABA transporters in atropine-treated myopic retina as revealed by iTRAQ quantitative proteomics.
Involvement of GABA transporters in atropine-treated myopic retina as revealed by iTRAQ quantitative proteomics.
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DOI：
10.1021/pr500558y
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发表时间：
2014-11-07
影响因子：
4.4
通讯作者：
Zhou, Lei
中科院分区：
文献类型：
--
作者：
Barathi, Veluchamy A.;Chaurasia, Shyam S.;Poidinger, Michael;Koh, Siew Kwan;Tian, Dechao;Ho, Candice;Iuvone, P. Michael;Beuerman, Roger W.;Zhou, Lei
阿托品是一种毒蕈碱拮抗剂，已知在几种动物模型和人类中抑制近视进展。然而，诉讼方式尚未确定。在这项研究中，我们比较了用阿托品治疗的对照和晶状体性近视（LIM）小鼠视网膜的iTRAQ定量蛋白质组学分析。近视组于出生后第10天配戴右眼（- 15D）镜片，从出生后第24天开始滴或不滴阿托品滴眼液。在出生后24、38和52天，通过光学低相干干涉测量（OLCI）和AC-Master测量轴向长度，并通过自动红外折射仪测量折射率。每组取6只眼睛的视网膜组织样本。实验重复2次，并进行液相色谱-串联质谱（LC-MS /MS）分析技术重复。使用MetaCore进行蛋白质谱分析以进行途径分析。通过对两个独立实验的重复分析，我们共鉴定出3882个FDR <1%的独特蛋白。这是迄今为止报道的数量最多的小鼠视网膜蛋白质组。与对照组相比，近视眼中有30个蛋白表达上调（近视/对照之比>全球平均比+ 1个标准差），28个蛋白表达下调（近视/对照之比<全球平均比- 1个标准差）。MetaCore通路分析揭示了近视眼中γ-氨基丁酸（GABA）水平的调节。详细的定量蛋白质组学数据分析显示，近视视网膜中GABA转运蛋白1 （GAT-1）水平升高，阿托品治疗后显著降低。免疫组织化学和Western blot分析进一步验证了这些结果。总之，本研究对阿托品处理的小鼠视网膜进行了全面的定量蛋白质组学分析，并提示gaba能信号参与了阿托品对小鼠眼睛的抗近视作用。gaba能在神经视网膜中的传递在哺乳动物轴眼生长的维持中起着关键作用。
Atropine, a muscarinic antagonist, is known to inhibit myopia progression in several animal models and humans. However, the mode of action is not established yet. In this study, we compared quantitative iTRAQ proteomic analysis in the retinas collected from control and lens-induced myopic (LIM) mouse eyes treated with atropine. The myopic group received a (−15D) spectacle lens over the right eye on postnatal day 10 with or without atropine eye drops starting on postnatal day 24. Axial length was measured by optical low coherence interferometry (OLCI), AC-Master, and refraction was measured by automated infrared photorefractor at postnatal 24, 38, and 52 days. Retinal tissue samples were pooled from six eyes for each group. The experiments were repeated twice, and technical replicates were also performed for liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. MetaCore was used to perform protein profiling for pathway analysis. We identified a total of 3882 unique proteins with <1% FDR by analyzing the samples in replicates for two independent experiments. This is the largest number of mouse retina proteome reported to date. Thirty proteins were found to be up-regulated (ratio for myopia/control > global mean ratio + 1 standard deviation), and 28 proteins were down-regulated (ratio for myopia/control < global mean ratio - 1 standard deviation) in myopic eyes as compared with control retinas. Pathway analysis using MetaCore revealed regulation of γ-aminobutyric acid (GABA) levels in the myopic eyes. Detailed analysis of the quantitative proteomics data showed that the levels of GABA transporter 1 (GAT-1) were elevated in myopic retina and significantly reduced after atropine treatment. These results were further validated with immunohistochemistry and Western blot analysis. In conclusion, this study provides a comprehensive quantitative proteomic analysis of atropine-treated mouse retina and suggests the involvement of GABAergic signaling in the antimyopic effects of atropine in mouse eyes. The GABAergic transmission in the neural retina plays a pivotal role in the maintenance of axial eye growth in mammals.
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DOI：
10.1124/jpet.108.146464
发表时间：
2009-02
影响因子：
3.5
作者：
Chebib, Mary;Hinton, Tina;Schmid, Katrina L.;Brinkworth, Darren;Qian, Haohua;Matos, Susana;Kim, Hye-Lim;Abdel-Halim, Heba;Kumar, Rohan J.;Johnston, Graham A. R.;Hanrahan, Jane R.
通讯作者：
Hanrahan, Jane R.
影响因子：
3.3
作者：
Gibbs, ME;Johnston, GAR
通讯作者：
Johnston, GAR
影响因子：
4.3
作者：
Barathi VA;Kwan JL;Tan QS;Weon SR;Seet LF;Goh LK;Vithana EN;Beuerman RW
通讯作者：
Beuerman RW
影响因子：
56.9
作者：
Fagiolini, M;Fritschy, JM;Hensch, TK
通讯作者：
Hensch, TK
影响因子：
1.4
作者：
Chia, Audrey;Li, Wen;Luu, Chi D.
通讯作者：
Luu, Chi D.