Department of Development and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY, 10461, USA.
Institute for Aging Studies, Albert Einstein College of Medicine, Bronx, NY, 10461, USA.
Nat Commun. 2020 Jan 31;11(1):645. doi: 10.1038/s41467-019-14164-4.
Autophagy malfunctioning occurs in multiple human disorders, making attractive the idea of chemically modulating it with therapeutic purposes. However, for many types of autophagy, a clear understanding of tissue-specific differences in their activity and regulation is missing because of lack of methods to monitor these processes in vivo. Chaperone-mediated autophagy (CMA) is a selective type of autophagy that until now has only been studied in vitro and not in the tissue context at single cell resolution. Here, we develop a transgenic reporter mouse that allows dynamic measurement of CMA activity in vivo using image-based procedures. We identify previously unknown spatial and temporal differences in CMA activity in multiple organs and in response to stress. We illustrate the versatility of this model for monitoring CMA in live animals, organotypic cultures and cell cultures from these mice, and provide practical examples of multiorgan response to drugs that modulate CMA.
自噬功能障碍发生在多种人类疾病中，这使得用化学方法调节自噬以达到治疗目的的想法具有吸引力。然而，由于缺乏监测这些过程在体内的方法，对于许多类型的自噬，它们在活性和调节方面的组织特异性差异仍不清楚。伴侣介导的自噬（CMA）是一种选择性的自噬，到目前为止，它只在体外进行过研究，而不是在单细胞分辨率的组织环境中进行研究。在这里，我们开发了一种转基因报告小鼠，允许使用基于图像的程序在体内动态测量 CMA 活性。我们在多个器官中发现了 CMA 活性以前未知的空间和时间差异，并对其做出了应激反应。我们说明了该模型在监测活体动物、器官型培养物和来自这些小鼠的细胞培养物中的 CMA 的多功能性，并提供了多器官对调节 CMA 的药物反应的实际示例。
