Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase
Ischemic episodes are a leading cause of death worldwide with limited therapeutic interventions. The current study explored mitochondrial phosphate-activated glutaminase (GLS1) activity modulation by PKCβII through GC-MS untargeted metabolomics approach. Mitochondria were used to elucidate the endog...
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doaj-81bda0b7600e4f678dee1f88ead3c7862021-08-26T13:51:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-01228504850410.3390/ijms22168504Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated GlutaminaseMałgorzata Beręsewicz-Haller0Olga Krupska1Paweł Bochomulski2Danuta Dudzik3Anita Chęcińska4Wojciech Hilgier5Coral Barbas6Krzysztof Zablocki7Barbara Zablocka8Molecular Biology Unit, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandMolecular Biology Unit, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandMolecular Biology Unit, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandCentre for Metabolomics and Bioanalysis (CEMBIO), Department of Chemistry and Biochemistry, Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, 28003 Madrid, SpainMolecular Biology Unit, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandDepartment of Neurotoxicology, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandCentre for Metabolomics and Bioanalysis (CEMBIO), Department of Chemistry and Biochemistry, Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, 28003 Madrid, SpainLaboratory of Cellular Metabolism, Nencki Institute of Experimental Biology, PAS, 02-106 Warsaw, PolandMolecular Biology Unit, Mossakowski Medical Research Institute, PAS, 02-106 Warsaw, PolandIschemic episodes are a leading cause of death worldwide with limited therapeutic interventions. The current study explored mitochondrial phosphate-activated glutaminase (GLS1) activity modulation by PKCβII through GC-MS untargeted metabolomics approach. Mitochondria were used to elucidate the endogenous resistance of hippocampal CA2-4 and dentate gyrus (DG) to transient ischemia and reperfusion in a model of ischemic episode in gerbils. In the present investigation, male gerbils were subjected to bilateral carotids occlusion for 5 min followed by reperfusion (IR). Gerbils were randomly divided into three groups as vehicle-treated sham control, vehicle-treated IR and PKCβII specific inhibitor peptide βIIV5-3-treated IR. Vehicle or βIIV5-3 (3 mg/kg, i.v.) were administered at the moment of reperfusion. The gerbils hippocampal tissue were isolated at various time of reperfusion and cell lysates or mitochondria were isolated from CA1 and CA2-4,DG hippocampal regions. Recombinant proteins PKCβII and GLS1 were used in in vitro phosphorylation reaction and organotypic hippocampal cultures (OHC) transiently exposed to NMDA (25 μM) to evaluate the inhibition of GLS1 on neuronal viability. PKCβII co-precipitates with GAC (GLS1 isoform) in CA2-4,DG mitochondria and phosphorylates GLS1 in vitro. Cell death was dose dependently increased when GLS1 was inhibited by BPTA while inhibition of mitochondrial pyruvate carrier (MPC) attenuated cell death in NMDA-challenged OHC. Fumarate and malate were increased after IR 1h in CA2-4,DG and this was reversed by βIIV5-3 what correlated with GLS1 activity increases and earlier showed elevation of neuronal death (Krupska et al., 2017). The present study illustrates that CA2-4,DG resistance to ischemic episode at least partially rely on glutamine and glutamate utilization in mitochondria as a source of carbon to tricarboxylic acid cycle. This phenomenon depends on modulation of GLS1 activity by PKCβII and remodeling of MPC: all these do not occur in ischemia-vulnerable CA1.https://www.mdpi.com/1422-0067/22/16/8504cerebral ischemiaendogenous neuroprotectionmitochondriaglutamate metabolismmetabolomicsprotein kinase C |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Małgorzata Beręsewicz-Haller Olga Krupska Paweł Bochomulski Danuta Dudzik Anita Chęcińska Wojciech Hilgier Coral Barbas Krzysztof Zablocki Barbara Zablocka |
spellingShingle |
Małgorzata Beręsewicz-Haller Olga Krupska Paweł Bochomulski Danuta Dudzik Anita Chęcińska Wojciech Hilgier Coral Barbas Krzysztof Zablocki Barbara Zablocka Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase International Journal of Molecular Sciences cerebral ischemia endogenous neuroprotection mitochondria glutamate metabolism metabolomics protein kinase C |
author_facet |
Małgorzata Beręsewicz-Haller Olga Krupska Paweł Bochomulski Danuta Dudzik Anita Chęcińska Wojciech Hilgier Coral Barbas Krzysztof Zablocki Barbara Zablocka |
author_sort |
Małgorzata Beręsewicz-Haller |
title |
Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase |
title_short |
Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase |
title_full |
Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase |
title_fullStr |
Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase |
title_full_unstemmed |
Mitochondrial Metabolism behind Region-Specific Resistance to Ischemia-Reperfusion Injury in Gerbil Hippocampus. Role of PKCβII and Phosphate-Activated Glutaminase |
title_sort |
mitochondrial metabolism behind region-specific resistance to ischemia-reperfusion injury in gerbil hippocampus. role of pkcβii and phosphate-activated glutaminase |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-08-01 |
description |
Ischemic episodes are a leading cause of death worldwide with limited therapeutic interventions. The current study explored mitochondrial phosphate-activated glutaminase (GLS1) activity modulation by PKCβII through GC-MS untargeted metabolomics approach. Mitochondria were used to elucidate the endogenous resistance of hippocampal CA2-4 and dentate gyrus (DG) to transient ischemia and reperfusion in a model of ischemic episode in gerbils. In the present investigation, male gerbils were subjected to bilateral carotids occlusion for 5 min followed by reperfusion (IR). Gerbils were randomly divided into three groups as vehicle-treated sham control, vehicle-treated IR and PKCβII specific inhibitor peptide βIIV5-3-treated IR. Vehicle or βIIV5-3 (3 mg/kg, i.v.) were administered at the moment of reperfusion. The gerbils hippocampal tissue were isolated at various time of reperfusion and cell lysates or mitochondria were isolated from CA1 and CA2-4,DG hippocampal regions. Recombinant proteins PKCβII and GLS1 were used in in vitro phosphorylation reaction and organotypic hippocampal cultures (OHC) transiently exposed to NMDA (25 μM) to evaluate the inhibition of GLS1 on neuronal viability. PKCβII co-precipitates with GAC (GLS1 isoform) in CA2-4,DG mitochondria and phosphorylates GLS1 in vitro. Cell death was dose dependently increased when GLS1 was inhibited by BPTA while inhibition of mitochondrial pyruvate carrier (MPC) attenuated cell death in NMDA-challenged OHC. Fumarate and malate were increased after IR 1h in CA2-4,DG and this was reversed by βIIV5-3 what correlated with GLS1 activity increases and earlier showed elevation of neuronal death (Krupska et al., 2017). The present study illustrates that CA2-4,DG resistance to ischemic episode at least partially rely on glutamine and glutamate utilization in mitochondria as a source of carbon to tricarboxylic acid cycle. This phenomenon depends on modulation of GLS1 activity by PKCβII and remodeling of MPC: all these do not occur in ischemia-vulnerable CA1. |
topic |
cerebral ischemia endogenous neuroprotection mitochondria glutamate metabolism metabolomics protein kinase C |
url |
https://www.mdpi.com/1422-0067/22/16/8504 |
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