Cytosolic LC3 ratio as a sensitive index of macroautophagy in isolated rat hepatocytes and H4-II-E cells

被引：108
作者：
Karim, Md. Razaul
Kanazawa, Takumi
Daigaku, Yasuhiro
Fujimura, Shinobu
Miotto, Giovanni
Kadowaki, Motoni
机构：
[1] Niigata Univ, Fac Agr, Dept Appl Biol Chem, Niigata 9502181, Japan
[2] Univ Padua, Dept Biol Chem, Padua, Italy
来源：
AUTOPHAGY | 2007年 / 3卷 / 06期
关键词：
amino acids; hepatocytes; H4-II-E cells; LC3; macroautophagy; proteolysis;
D O I：
10.4161/auto.4615
中图分类号：
Q2 [细胞生物学];
学科分类号：
071009 ; 090102 ;
摘要：
Macroautophagy, an intracellular bulk degradation process in eukaryotes, is sensitive to nutrient supply and deprivation. Microtubule-associated protein 1 light chain 3 (LC3), a mammalian homologue of yeast Atg8, plays an indispensable role in macroautophagy formation and is a suitable marker for this process. Through analysis of the subcellular distribution of LC3, we determined that the cytosolic fraction contained not only a precursor form (LC3-I), but also an apparent active form (LC3-IIs). Both cytosolic LC3-I and LC3-IIs were more responsive to amino acids than those of total homogenate. Moreover, changes in the LC3-IIs/I ratio reflected those in the total proteolytic flux remarkably in both fresh rat hepatocytes and H4-II-E cell lines. Thus, in addition to a sensitive index of macroautophagy, calculating the cytosolic LC3 ratio became an easy and quick quantitative method for monitoring its regulation in hepatocytes and H4-II-E cells.
引用
收藏
页码：553 / 560
页数：8
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