Global ischemia induces lysosomal-mediated degradation of mTOR and activation of autophagy in hippocampal neurons destined to die

Cell Death Differ. 2017 Feb;24(2):317-329. doi: 10.1038/cdd.2016.140. Epub 2016 Dec 9.

Abstract

The mammalian target of rapamycin (mTOR) is a key regulator of cell growth, autophagy, translation, and survival. Dysregulation of mTOR signaling is associated with cancer, diabetes, and autism. However, a role for mTOR signaling in neuronal death is not well delineated. Here we show that global ischemia triggers a transient increase in mTOR phosphorylation at S2448, whereas decreasing p-mTOR and functional activity in selectively vulnerable hippocampal CA1 neurons. The decrease in mTOR coincides with an increase in biochemical markers of autophagy, pS317-ULK-1, pS14-Beclin-1, and LC3-II, a decrease in the cargo adaptor p62, and an increase in autophagic flux, a functional readout of autophagy. This is significant in that autophagy, a catabolic process downstream of mTORC1, promotes the formation of autophagosomes that capture and target cytoplasmic components to lysosomes. Inhibitors of the lysosomal (but not proteasomal) pathway rescued the ischemia-induced decrease in mTOR, consistent with degradation of mTOR via the autophagy/lysosomal pathway. Administration of the mTORC1 inhibitor rapamycin or acute knockdown of mTOR promotes autophagy and attenuates ischemia-induced neuronal death, indicating an inverse causal relation between mTOR, autophagy, and neuronal death. Our findings identify a novel and previously unappreciated mechanism by which mTOR self-regulates its own levels in hippocampal neurons in a clinically relevant model of ischemic stroke.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Acetylcysteine / analogs & derivatives
  • Acetylcysteine / pharmacology
  • Adenine / analogs & derivatives
  • Adenine / pharmacology
  • Animals
  • Autophagy* / drug effects
  • Autophagy-Related Protein-1 Homolog / metabolism
  • Beclin-1 / metabolism
  • Cells, Cultured
  • Hippocampus / cytology
  • Ischemia / metabolism
  • Ischemia / pathology
  • Leupeptins / pharmacology
  • Lysosomes / drug effects
  • Lysosomes / metabolism*
  • Male
  • Microtubule-Associated Proteins / metabolism
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism
  • Phosphorylation / drug effects
  • RNA Interference
  • Rats
  • Sirolimus / pharmacology
  • TOR Serine-Threonine Kinases / antagonists & inhibitors
  • TOR Serine-Threonine Kinases / genetics
  • TOR Serine-Threonine Kinases / metabolism*

Substances

  • Beclin-1
  • Leupeptins
  • Microtubule-Associated Proteins
  • lactacystin
  • 3-methyladenine
  • Autophagy-Related Protein-1 Homolog
  • TOR Serine-Threonine Kinases
  • AMP-Activated Protein Kinases
  • leupeptin
  • Adenine
  • Sirolimus
  • Acetylcysteine