Hato Takashi, Sandoval Ruben, Dagher Pierre C
Department of Medicine; Indiana University; Indianapolis, IN USA.
Intravital. 2015;4(2). doi: 10.1080/21659087.2015.1067352. Epub 2015 Aug 28.
Tubular cell apoptosis is a major phenotype of cell death in various forms of acute kidney injury. Quantifying apoptosis in fixed tissues is problematic because apoptosis evolves over time and dead cells are rapidly cleared by the phagocytic system. Phiphilux is a fluorescent probe that is activated specifically by caspase 3 and does not inhibit the subsequent activity of this effector caspase. It has been used successfully to quantify apoptosis in cell culture. Here we examined the feasibility of using Phiphilux to measure renal tubular apoptosis progression over time in live animals using intravital 2-photon microscopy. Our results show that Phiphilux can detect apoptosis in S2 tubules but is activated non-specifically in S1 tubules.
肾小管细胞凋亡是各种形式急性肾损伤中细胞死亡的主要表型。对固定组织中的凋亡进行定量存在问题,因为凋亡会随时间演变,且死亡细胞会被吞噬系统迅速清除。Phiphilux是一种荧光探针,它被半胱天冬酶3特异性激活,且不抑制这种效应半胱天冬酶的后续活性。它已成功用于细胞培养中凋亡的定量。在此,我们使用活体双光子显微镜检查了使用Phiphilux测量活体动物肾小管凋亡随时间进展的可行性。我们的结果表明,Phiphilux可以检测S2段肾小管中的凋亡,但在S1段肾小管中被非特异性激活。