Ly Alice, Merl-Pham Juliane, Priller Markus, Gruhn Fabian, Senninger Nicole, Ueffing Marius, Hauck Stefanie M
Research Unit Protein Science, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH) , Neuherberg, D-85764 Germany.
Research Unit Analytical Pathology, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH) , Neuherberg, D-85764 Germany.
J Proteome Res. 2016 Apr 1;15(4):1350-9. doi: 10.1021/acs.jproteome.6b00111. Epub 2016 Mar 14.
The rd10 mouse is a model of retinitis pigmentosa characterized by the dysfunction of a rod-photoreceptor-specific phosphodiesterase. Compared to the rd1 mouse, retinal degeneration in the rd10 mouse begins later in age with a milder phenotype, making it ideal for investigating cell death and neuroprotective mechanisms. Alterations in the rd10 retina proteome at pre-, peak-, and postdegenerative time points were examined using a modified high-recovery filter-aided sample preparation (FASP) method in combination with label-free quantitative mass spectrometry, generating a proteomic data set on almost 3000 proteins. Our data confirmed a period of protein expression similar to age-matched wild-type mice predegeneration, with decreases in proteins associated with phototransduction and increases in signaling proteins at peak- and postdegenerative stages. A total of 57 proteins were differentially expressed in the rd10 retinae during peak-degeneration, compared to those in wild-type mice after stringent FDR correction (q < 0.05). Network analysis separated these proteins into one cluster of down-regulated photoreceptor proteins and one of up-regulated signaling proteins centered around GFAP, STAT3, and STAT1. This is the first study to identify alterations in STAT1 in the rd10 mouse, which were confirmed with gene expression and immunoblotting experiments, underpinning the efficacy of our approach. This unique proteomic data set on protein dynamics during retinal degeneration could serve as an information source for vision research in the future.
rd10小鼠是一种视网膜色素变性模型，其特征在于视杆光感受器特异性磷酸二酯酶功能障碍。与rd1小鼠相比，rd10小鼠的视网膜变性在年龄较大时开始，表型较轻，这使其成为研究细胞死亡和神经保护机制的理想模型。使用改良的高回收率滤膜辅助样品制备（FASP）方法结合无标记定量质谱法，检测了rd10视网膜蛋白质组在变性前、高峰期和变性后的时间点的变化，生成了一个包含近3000种蛋白质的蛋白质组数据集。我们的数据证实，在变性前，蛋白质表达期与年龄匹配的野生型小鼠相似，在变性高峰期和变性后，与光转导相关的蛋白质减少，信号蛋白增加。与野生型小鼠相比，在严格的FDR校正后（q < 0.05），共有57种蛋白质在rd10视网膜变性高峰期差异表达。网络分析将这些蛋白质分为一组下调的光感受器蛋白质和一组以上调的信号蛋白为中心的蛋白质，这些信号蛋白以GFAP、STAT3和STAT1为中心。这是第一项鉴定rd10小鼠中STAT1变化的研究，基因表达和免疫印迹实验证实了这一点，支持了我们方法的有效性。这个关于视网膜变性期间蛋白质动态变化的独特蛋白质组数据集可以作为未来视觉研究的信息来源。
