Laboratory of Experimental Optometry, Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, Kowloon, Hong Kong.
Laboratory of Experimental Optometry, Centre for Myopia Research, School of Optometry, The Hong Kong Polytechnic University, Kowloon, Hong Kong; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, China.
J Proteomics. 2018 Jun 15;181:1-15. doi: 10.1016/j.jprot.2018.03.023. Epub 2018 Mar 20.
Myopia is generally regarded as a failure of normal emmetropization process, however, its underlying molecular mechanisms are unclear. To investigate the retinal protein profile changes during emmetropization, we studied differential protein expressions of ocular growth in young guinea pigs at 3 and 21 days old respectively, when significant axial elongation was detected (P < 0.001, n = 10). Independent pooled retinal samples of both eyes were subjected to SWATH mass spectrometry (MS) followed by bioinformatics analysis using cloud-based platforms. A comprehensive retina SWATH ion-library consisting of 3138 (22,871) unique proteins (peptides) at 1% FDR was constructed. 40 proteins were found to be significantly up-regulated and 8 proteins down-regulated during emmetropization (≥log2 of 0.43 with ≥2 peptides matched per protein; P < 0.05). Using pathway analysis, the most significant pathway identifiable was 'phototransduction' (P = 1.412e-4). Expression patterns of 7 proteins identified in this pathway were further validated and confirmed (P < 0.05) with high-resolution Multiple Reaction Monitoring (MRM-HR) MS. Combining discovery and targeted proteomics approaches, this study for the first time comprehensively profiled protein changes in the guinea pig retina during normal emmetropization-associated eye growth. The findings of this study are also relevant to the myopia development, which is the result of failed emmetropization.
Myopia is considered as a failure of emmetropization. However, the underlying biochemical mechanism of emmetropization, a visually guided process in which eye grows towards the optimal optical state of clear vision during early development, is not well understood. Retina is known as the key tissue to regulate this active eye growth. we studied eye growth of young guinea pigs and harvested their retinal tissues. A comprehensive SWATH ion library with identification of a total 3138 unique proteins were established, in which 48 proteins exhibited significant differential expressions between 3 and 21 days old. After MRM-HR confirmation, 'phototransduction' were found as the most active pathway during emmetropic eye growth. This study is the first in discovering key retinal protein players and pathways which are presumably orchestrated by biological mechanism(s) underlying emmetropization.
近视通常被认为是正视化过程的失败，然而，其潜在的分子机制尚不清楚。为了研究正视化过程中视网膜蛋白谱的变化，我们分别研究了 3 天和 21 天龄的幼豚鼠眼球生长的差异蛋白表达，此时明显的轴向伸长被检测到（P<0.001，n=10）。将双眼独立的 pooled 视网膜样本进行 SWATH 质谱（MS）分析，并使用基于云的平台进行生物信息学分析。构建了一个由 3138（22871）个独特蛋白（肽）组成的全面视网膜 SWATH 离子文库，假阳性率为 1%。发现 40 种蛋白在正视化过程中显著上调，8 种蛋白下调（≥log2 的 0.43，每种蛋白有≥2 个肽匹配；P<0.05）。通过通路分析，可识别的最显著通路是“光转导”（P=1.412e-4）。该通路中鉴定的 7 种蛋白的表达模式进一步用高分辨率多重反应监测（MRM-HR）MS 进行验证和确认（P<0.05）。本研究首次采用发现和靶向蛋白质组学方法，全面分析了正常正视相关眼生长过程中豚鼠视网膜的蛋白质变化。该研究的发现也与近视的发展有关，近视是正视化失败的结果。
近视被认为是正视化的失败。然而，作为一个视觉引导的过程，眼球在早期发育过程中朝着清晰视觉的最佳光学状态生长，其背后的生化机制尚不清楚。视网膜被认为是调节这种主动眼球生长的关键组织。我们研究了幼豚鼠的眼球生长，并采集了它们的视网膜组织。建立了一个具有 3138 个独特蛋白鉴定的全面 SWATH 离子文库，其中 3 天和 21 天龄之间有 48 种蛋白表现出显著的差异表达。经 MRM-HR 确认后，发现“光转导”是正视性眼球生长中最活跃的通路。这项研究首次发现了可能由正视化背后的生物学机制协调的关键视网膜蛋白和通路。
