Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells.
2020
Authors:
Vučićević, LjubicaMisirkić Marjanović, Maja
Ćirić, Darko
Martinović, Tamara
Jovanović, Maja
Isaković, Aleksandra
Marković, Ivanka
Šaponjić, Jasna
Foretz, Marc
Rabanal-Ruiz, Yoana
Korolchuk, Viktor I.
Trajković, Vladimir
Document Type:
Article (Published version)
,
© 2019, Springer Nature Switzerland AG.
Metadata
Show full item recordAbstract:
We investigated the role of autophagy, a controlled lysosomal degradation of cellular macromolecules and organelles, in glutamate excitotoxicity during nutrient deprivation in vitro. The incubation in low-glucose serum/amino acid-free cell culture medium synergized with glutamate in increasing AMP/ATP ratio and causing excitotoxic necrosis in SH-SY5Y human neuroblastoma cells. Glutamate suppressed starvation-triggered autophagy, as confirmed by diminished intracellular acidification, lower LC3 punctuation and LC3-I conversion to autophagosome-associated LC3-II, reduced expression of proautophagic beclin-1 and ATG5, increase of the selective autophagic target NBR1, and decreased number of autophagic vesicles. Similar results were observed in PC12 rat pheochromocytoma cells. Both glutamate-mediated excitotoxicity and autophagy inhibition in starved SH-SY5Y cells were reverted by NMDA antagonist memantine and mimicked by NMDA agonists D-aspartate and ibotenate. Glutamate reduced starvation-triggered phosphorylation of the energy sensor AMP-activated protein kinase (AMPK) without affecting the activity of mammalian target of rapamycin complex 1, a major negative regulator of autophagy. This was associated with reduced mRNA levels of autophagy transcriptional activators (FOXO3, ATF4) and molecules involved in autophagy initiation (ULK1, ATG13, FIP200), autophagosome nucleation/elongation (ATG14, beclin-1, ATG5), and autophagic cargo delivery to autophagosomes (SQSTM1). Glutamate-mediated transcriptional repression of autophagy was alleviated by overexpression of constitutively active AMPK. Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, as well as genetic or pharmacological autophagy induction by TFEB overexpression or lithium chloride, reduced the sensitivity of nutrient-deprived SH-SY5Y cells to glutamate excitotoxicity. These data indicate that transcriptional inhibition of AMPK-dependent cytoprotective autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.
Keywords:
Brain; Energy stress; Ischemia; Neurodegeneration; NeurotoxicitySource:
Cellular and Molecular Life Sciences, 2020, 77, 3383-3399Funding / projects:
- The role of autophagy in regulation of cancer cell death (RS-MESTD-Basic Research (BR or ON)-173053)
- Modulation of intracellular energy balance-controlling signalling pathways in therapy of cancer and neuro-immuno-endocrine disorders (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41025)
- Neurobiology of sleep in aging and disease - electroencephalographic markers and modeling in the estimation of disorder (RS-MESTD-Basic Research (BR or ON)-173022)
- Cost Action TRANSAUTOPHAGY CA15138
DOI: 10.1007/s00018-019-03356-2
ISSN: 1420-682X
PubMed: 31720741
WoS: 000495944900001
Scopus: 2-s2.0-85075142282
URI
http://link.springer.com/10.1007/s00018-019-03356-2https://radar.ibiss.bg.ac.rs/handle/123456789/3528