Tracking the fate of glomerular epithelial cells in vivo using serial multiphoton imaging in new mouse models with fluorescent lineage tags.

Tracking the fate of glomerular epithelial cells in vivo using serial multiphoton imaging in new mouse models with fluorescent lineage tags.
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在带有荧光谱系标签的新小鼠模型中,使用串行多人成像在体内跟踪肾小球上皮细胞的命运。

DOI:
10.1038/nm.3405
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发表时间:
2013-12
期刊:
影响因子:
82.9
通讯作者:
--
中科院分区:
医学1区
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--
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足细胞对维持健康的肾小球滤器至关重要,但由于技术限制,在完整的肾脏中对足细胞进行研究一直是困难的。在这里,我们报道了连续几天对同一肾小球的多光子显微镜(MPM)的发展,以显示体内足细胞和壁上皮细胞(PEC)的运动。在Podocin-GFP小鼠中,足细胞在单侧输尿管结扎(UUO)后形成零星的多细胞簇,并迁移到顶鲍曼囊中。以CFP、GFP、YFP或RFP的细胞特异性表达为特征的Podocin-五彩纸屑小鼠中单个细胞的跟踪表明,多个足细胞同时迁移。在PEPCK-GFP小鼠中,序列MPM发现了PEC到足细胞的迁移和纳米管连接。我们的数据支持肾小球环境和细胞组成的高度动态而不是静态的性质。这一新方法的未来应用有望促进我们对肾小球损伤和再生机制的理解。
Podocytes are critical in the maintenance of a healthy glomerular filter, however they have been difficult to study in the intact kidney due to technical limitations. Here we report the development of serial multiphoton microscopy (MPM) of the same glomeruli over several days to visualize the motility of podocytes and parietal epithelial cells (PEC) in vivo. In Podocin-GFP mice podocytes formed sporadic multi-cellular clusters after unilateral ureteral ligation (UUO) and migrated into the parietal Bowman’s capsule. The tracking of single cells in Podocin-confetti mice featuring cell-specific expression of CFP, GFP, YFP, or RFP revealed the simultaneous migration of multiple podocytes. In PEPCK-GFP mice serial MPM found PEC-to-podocyte migration and nanotubule connections. Our data support the highly dynamic rather than static nature of the glomerular environment and cellular composition. Future application of this new approach promises to advance our understanding of the mechanisms of glomerular injury and regeneration.
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