Rocchi Altea, He Congcong
Department of Cell and Molecular Biology, Northwestern University.
Department of Cell and Molecular Biology, Northwestern University;
J Vis Exp. 2017 Feb 3(120):55099. doi: 10.3791/55099.
Autophagy is a lysosomal degradation pathway essential for cell homeostasis, function and differentiation. Under stress conditions, autophagy is induced and targets various cargos, such as bulk cytosol, damaged organelles and misfolded proteins, for degradation in lysosomes. Resulting nutrient molecules are recycled back to the cytosol for new protein synthesis and ATP production. Upregulation of autophagy has beneficial effects against the pathogenesis of many diseases, and pharmacological and physiological strategies to activate autophagy have been reported. Aerobic exercise is recently identified as an efficient autophagy inducer in multiple organs in mice, including muscle, liver, heart and brain. Here we show procedures to induce autophagy in vivo by either forced treadmill exercise or voluntary wheel running. We also demonstrate microscopic and biochemical methods to quantitatively analyze autophagy levels in mouse tissues, using the marker proteins LC3 and p62 that are transported to and degraded in lysosomes along with autophagosomes.
自噬是一种对细胞稳态、功能和分化至关重要的溶酶体降解途径。在应激条件下,自噬被诱导并靶向各种货物,如大量胞质溶胶、受损细胞器和错误折叠的蛋白质,以便在溶酶体中进行降解。产生的营养分子被循环回胞质溶胶用于新的蛋白质合成和ATP生成。自噬的上调对许多疾病的发病机制具有有益作用,并且已经报道了激活自噬的药理学和生理学策略。有氧运动最近被确定为小鼠多个器官(包括肌肉、肝脏、心脏和大脑)中的一种有效的自噬诱导剂。在这里,我们展示了通过强制跑步机运动或自愿轮转跑步在体内诱导自噬的程序。我们还展示了使用标记蛋白LC3和p62对小鼠组织中的自噬水平进行定量分析的显微镜和生化方法,这些标记蛋白会与自噬体一起转运到溶酶体中并在其中降解。