Neurobiology, Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
Institute of Biomedical Sciences, George Washington University, Washington, District of Columbia, United States of America.
PLoS Genet. 2020 Dec 23;16(12):e1009259. doi: 10.1371/journal.pgen.1009259. eCollection 2020 Dec.
Rab-GTPases and associated effectors mediate cargo transport through the endomembrane system of eukaryotic cells, regulating key processes such as membrane turnover, signal transduction, protein recycling and degradation. Using developmental transcriptome data, we identified Rabgef1 (encoding the protein RabGEF1 or Rabex-5) as the only gene associated with Rab GTPases that exhibited strong concordance with retinal photoreceptor differentiation. Loss of Rabgef1 in mice (Rabgef1-/-) resulted in defects specifically of photoreceptor morphology and almost complete loss of both rod and cone function as early as eye opening; however, aberrant outer segment formation could only partly account for visual function deficits. RabGEF1 protein in retinal photoreceptors interacts with Rabaptin-5, and RabGEF1 absence leads to reduction of early endosomes consistent with studies in other mammalian cells and tissues. Electron microscopy analyses reveal abnormal accumulation of macromolecular aggregates in autophagosome-like vacuoles and enhanced immunostaining for LC3A/B and p62 in Rabgef1-/- photoreceptors, consistent with compromised autophagy. Transcriptome analysis of the developing Rabgef1-/- retina reveals altered expression of 2469 genes related to multiple pathways including phototransduction, mitochondria, oxidative stress and endocytosis, suggesting an early trajectory of photoreceptor cell death. Our results implicate an essential role of the RabGEF1-modulated endocytic and autophagic pathways in photoreceptor differentiation and homeostasis. We propose that RabGEF1 and associated components are potential candidates for syndromic traits that include a retinopathy phenotype.
Rab-GTPases 和相关效应物介导真核细胞的内质网系统中的货物运输,调节关键过程,如膜周转、信号转导、蛋白质回收和降解。使用发育转录组数据,我们鉴定出 Rabgef1(编码蛋白 RabGEF1 或 Rabex-5)是唯一与 Rab GTPases 相关的基因,与视网膜光感受器分化具有很强的一致性。在小鼠中敲除 Rabgef1(Rabgef1-/-)会导致光感受器形态缺陷,并且早在眼睛张开时就几乎完全丧失了杆状和锥状功能;然而,异常的外节形成只能部分解释视觉功能缺陷。视网膜光感受器中的 RabGEF1 蛋白与 Rabaptin-5 相互作用,RabGEF1 的缺失导致早期内体的减少,这与其他哺乳动物细胞和组织的研究一致。电子显微镜分析显示,在自噬体样空泡中大分子聚集体异常积累,并且 Rabgef1-/-光感受器中的 LC3A/B 和 p62 的免疫染色增强,与自噬受损一致。对发育中的 Rabgef1-/-视网膜的转录组分析显示,与光转导、线粒体、氧化应激和内吞作用等多种途径相关的 2469 个基因的表达发生改变,表明光感受器细胞死亡的早期轨迹。我们的结果表明,RabGEF1 调节的内吞和自噬途径在光感受器分化和稳态中起着重要作用。我们提出 RabGEF1 和相关成分是包括视网膜病变表型在内的综合征特征的潜在候选者。