背景:先前的研究表明,在new的晶状体再生过程中存在巨噬细胞,但是他们在这个过程中的作用还有待阐明。
方法:在这里,我们使用new,PleurodelesWaltl,在晶状体再生过程中追踪巨噬细胞。此外,我们使用两种new物种评估晶状体再生过程中基因表达的早期变化,诺特眼为病毒体和胸膜虫。最后,我们在这两个物种的晶状体再生过程中使用了氯膦酸盐脂质体来消耗巨噬细胞,并测试了巨噬细胞恢复后后续继发性损伤的影响.
结果:巨噬细胞耗尽,晶状体再生,诱导瘢痕样组织的形成,导致炎症,减少虹膜色素上皮细胞(iPEC)增殖,和增加眼睛的凋亡率。这些表型中的一些在100天的最后观察期内持续存在,并且可以通过外源性FGF2施用来减毒。为背侧虹膜建立了编码急性炎症效应物的独特转录物图谱。new眼的再损伤减轻了巨噬细胞消耗的影响,包括疤痕样组织的分辨率,并重新开始再生过程。
结论:一起,我们的研究结果强调了巨噬细胞通过调节纤维化反应促进new眼再生环境的重要性,调节整体炎症环境,维持iPECs早期增殖和晚期凋亡的适当平衡。
BACKGROUND: Previous studies have suggested that macrophages are present during lens regeneration in newts, but their role in the process is yet to be elucidated.
METHODS: Here we generated a transgenic reporter line using the newt, Pleurodeles waltl, that traces macrophages during lens regeneration. Furthermore, we assessed early changes in gene expression during lens regeneration using two newt species, Notophthalmus viridescens and Pleurodeles waltl. Finally, we used clodronate liposomes to deplete macrophages during lens regeneration in both species and tested the effect of a subsequent secondary injury after macrophage recovery.
RESULTS: Macrophage depletion abrogated lens regeneration, induced the formation of scar-like tissue, led to inflammation, decreased iris pigment epithelial cell (iPEC) proliferation, and increased rates of apoptosis in the eye. Some of these phenotypes persisted throughout the last observation period of 100 days and could be attenuated by exogenous FGF2 administration. A distinct transcript profile encoding acute inflammatory effectors was established for the dorsal iris. Reinjury of the newt eye alleviated the effects of macrophage depletion, including the resolution of scar-like tissue, and re-initiated the regeneration process.
CONCLUSIONS: Together, our findings highlight the importance of macrophages for facilitating a pro-regenerative environment in the newt eye by regulating fibrotic responses, modulating the overall inflammatory landscape, and maintaining the proper balance of early proliferation and late apoptosis of the iPECs.