Qian Cheng, Xin Ying, Cheng Qi, Wang Hui, Zack Donald, Blackshaw Seth, Hattar Samer, Feng-Quan Zhou, Qian Jiang
Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine, Baltimore, United States.
Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, United States.
Res Sq. 2023 Aug 17:rs.3.rs-3193738. doi: 10.21203/rs.3.rs-3193738/v1.
The progressive death of mature neurons often results in neurodegenerative diseases. While the previous studies have mostly focused on identifying intrinsic mechanisms controlling neuronal survival, the extracellular environment also plays a critical role in regulating cell viability. Here we explore how intercellular communication contributes to the survival of retinal ganglion cells (RGCs) following the optic nerve crush (ONC). Although the direct effect of the ONC is restricted to the RGCs, we observed transcriptomic responses in other retinal cells to the injury based on the single-cell RNA-seq, with astrocytes and Müller glia having the most interactions with RGCs. By comparing the RGC subclasses showing distinct resilience to ONC-induced cell death, we found that the high-survival RGCs tend to have more ligand-receptor interactions with other retinal cells, suggesting that these RGCs are intrinsically programmed to foster more communication with their surroundings. Furthermore, we identified top 47 interactions that are stronger in the high-survival RGCs, likely representing neuroprotective interactions. We performed functional assays on one of the receptors, μ opioid receptor (Oprm1), a receptor known to play roles in regulating pain, reward, and addictive behavior. Although Oprm1 is preferentially expressed in intrinsically photosensitive retinal ganglion cells (ipRGCs), its neuroprotective effect could be transferred to multiple RGC subclasses by specific overexpressing Oprm1 in pan-RGCs in ONC, excitotoxicity, and glaucoma models. Lastly, manipulating Oprm1 activity improved visual functions and altered pupillary light response in mice. Our study provides an atlas of cell-cell interactions in both intact and post-ONC retina and an effective strategy to predict molecular mechanisms in neuroprotection, underlying the principal role played by extracellular environment in supporting neuron survival.
成熟神经元的渐进性死亡通常会导致神经退行性疾病。虽然先前的研究大多集中在确定控制神经元存活的内在机制,但细胞外环境在调节细胞活力方面也起着关键作用。在这里,我们探讨细胞间通讯如何在视神经挤压(ONC)后促进视网膜神经节细胞(RGC)的存活。尽管ONC的直接影响仅限于RGC,但基于单细胞RNA测序,我们观察到其他视网膜细胞对损伤的转录组反应,其中星形胶质细胞和穆勒胶质细胞与RGC的相互作用最多。通过比较对ONC诱导的细胞死亡表现出不同恢复力的RGC亚类,我们发现高存活率的RGC往往与其他视网膜细胞有更多的配体-受体相互作用,这表明这些RGC在本质上被编程为与周围环境进行更多的通讯。此外,我们确定了在高存活率的RGC中更强的前47种相互作用,可能代表神经保护相互作用。我们对其中一种受体μ阿片受体(Oprm1)进行了功能测定,该受体已知在调节疼痛、奖赏和成瘾行为中起作用。虽然Oprm1优先表达于内在光敏性视网膜神经节细胞(ipRGC)中,但其神经保护作用可以通过在ONC、兴奋性毒性和青光眼模型的全RGC中特异性过表达Oprm1转移到多个RGC亚类。最后,操纵Oprm1活性改善了小鼠的视觉功能并改变了瞳孔对光反应。我们的研究提供了完整和ONC后视网膜中细胞-细胞相互作用的图谱,以及一种预测神经保护分子机制的有效策略,强调了细胞外环境在支持神经元存活中的主要作用。