Ratiometric and discriminative visualization of autophagic processes with a novel dual-responded lysosome-specific fluorescent probe.

IF 11.3 1区 医学 Q1 Medicine Biomaterials Research Pub Date : 2023-07-06 DOI:10.1186/s40824-023-00409-3
Fan Zheng, Yeshuo Ma, Jipeng Ding, Shuai Huang, Shengwang Zhang, Xueyan Huang, Bin Feng, Hongliang Zeng, Fei Chen, Wenbin Zeng
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引用次数: 1

Abstract

Background: Autophagy is a critical self-eating pathway involved in numerous physiological and pathological processes. Lysosomal degradation of dysfunctional organelles and invading microorganisms is central to the autophagy mechanism and essential for combating disease-related conditions. Therefore, monitoring fluctuations in the lysosomal microenvironment is vital for tracking the dynamic process of autophagy. Although much effort has been put into designing probes for measuring lysosomal viscosity or pH separately, there is a need to validate the concurrent imaging of the two elements to enhance the understanding of the dynamic progression of autophagy.

Methods: Probe HFI was synthesized in three steps and was developed to visualize changes in viscosity and pH within lysosomes for real-time autophagy tracking. Then, the spectrometric determination was carried out. Next, the probe was applied to image autophagy in cells under nutrient-deprivation or external stress. Additionally, the performance of HFI to monitor autophagy was employed to evaluate acetaminophen-induced liver injury.

Results: We constructed a ratiometric dual-responsive probe, HFI, with a large Stokes shift over 200 nm, dual-wavelength emission, and small background interference. The ratiometric fluorescent signal (R = I 610/I 460) of HFI had an excellent correlation with both viscosity and pH. More importantly, high viscosity and low pH had a synergistic promotion effect on the emission intensity of HFI, which enabled it to specially lit lysosomes without disturbing the inherent microenvironment. We then successfully used HFI to monitor intracellular autophagy induced by starvation or drugs in real-time. Interestingly, HFI also enabled us to visualize the occurrence of autophagy in the liver tissue of a DILI model, as well as the reversible effect of hepatoprotective drugs on this event.

Conclusions: In this study, we developed the first ratiometric dual-responsive fluorescent probe, HFI, for real-time revealing autophagic details. It could image lysosomes with minimal perturbation to their inherent pH, allowing us to track changes in lysosomal viscosity and pH in living cells. Ultimately, HFI has great potential to serve as a useful indicator for autophagic changes in viscosity and pH in complex biological samples and can also be used to assess drug safety.

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一种新型双反应溶酶体特异性荧光探针对自噬过程的比例和判别可视化。
背景:自噬是一种重要的自食途径,参与许多生理和病理过程。功能失调细胞器和入侵微生物的溶酶体降解是自噬机制的核心,也是对抗疾病相关病症的必要条件。因此,监测溶酶体微环境的波动对于跟踪自噬的动态过程至关重要。尽管在设计分别测量溶酶体粘度或pH的探针方面已经付出了很多努力,但仍需要验证这两个元素的同时成像,以增强对自噬动态进展的理解。方法:探针HFI分三步合成,用于可视化溶酶体内粘度和pH的变化,实时跟踪自噬。然后进行光谱测定。接下来,应用探针成像细胞在营养剥夺或外部应激下的自噬。此外,采用HFI监测自噬的性能来评估对乙酰氨基酚诱导的肝损伤。结果:我们构建了一个比例双响应探针,HFI,在200 nm范围内具有大的Stokes位移,双波长发射,小背景干扰。HFI的比值荧光信号(R = I 610/I 460)与黏度和pH值均具有良好的相关性。更重要的是,高黏度和低pH值对HFI的发射强度有协同促进作用,使其能够在不干扰固有微环境的情况下特异性地照亮溶酶体。然后,我们成功地利用HFI实时监测饥饿或药物诱导的细胞内自噬。有趣的是,HFI还使我们能够可视化DILI模型肝组织中自噬的发生,以及肝保护药物对这一事件的可逆作用。结论:在这项研究中,我们开发了第一个比例双响应荧光探针HFI,用于实时显示自噬的细节。它可以在对溶酶体固有pH值进行最小扰动的情况下对溶酶体进行成像,使我们能够跟踪活细胞中溶酶体粘度和pH值的变化。最终,HFI具有很大的潜力,可作为复杂生物样品中粘度和pH自噬变化的有用指标,也可用于评估药物安全性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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