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

Md Razaul Karim, Takumi Kanazawa, Yasuhiro Daigaku, Shinobu Fujimura, Giovanni Miotto, Motoni Kadowaki

Research output: Contribution to journalArticlepeer-review

98 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)553-560
Number of pages8
JournalAutophagy
Volume3
Issue number6
DOIs
StatePublished - 2007
Externally publishedYes

Keywords

  • Amino acids
  • H4-II-E cells
  • Hepatocytes
  • LC3
  • Macroautophagy
  • Proteolysis

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