Global brain ischemia in rats is associated with mitochondrial release and downregulation of Mfn2 in the cerebral cortex, but not the hippocampus

Klacanova,Katarina; Kovalska,Maria; Chomova,Maria; Pilchova,Ivana; Tatarkova,Zuzana; Kaplan,Peter; Racay,Peter

doi:10.3892/ijmm.2019.4168

Global brain ischemia in rats is associated with mitochondrial release and downregulation of Mfn2 in the cerebral cortex, but not the hippocampus

Authors:
- Katarina Klacanova
- Maria Kovalska
- Maria Chomova
- Ivana Pilchova
- Zuzana Tatarkova
- Peter Kaplan
- Peter Racay
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Affiliations: Biomedical Center and Institute of Medical Biochemistry, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, SK‑03601 Martin, Slovak Republic, Department of Histology and Embryology, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, SK‑03601 Martin, Slovak Republic, Institute of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, Comenius University in Bratislava, SK‑81108 Bratislava, Slovak Republic, Department of Medical Biochemistry, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, SK‑03601 Martin, Slovak Republic
Published online on: April 16, 2019 https://doi.org/10.3892/ijmm.2019.4168
Pages: 2420-2428
Copyright: © Klacanova et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Mitochondria are crucial for neuronal cell survival and death through their functions in ATP production and the intrinsic pathway of apoptosis. Mitochondrial dysfunction is considered to play a central role in several serious human diseases, including neurodegenerative diseases, such as Parkinson's and Alzheimer's disease and ischemic neurodegeneration. The aim of the present study was to investigate the impact of transient global brain ischemia on the expression of selected proteins involved in mitochondrial dynamics and mitochondria‑associated membranes. The main foci of interest were the proteins mitofusin 2 (Mfn2), dynamin‑related protein 1 (DRP1), voltage‑dependent anion‑selective channel 1 (VDAC1) and glucose‑regulated protein 75 (GRP75). Western blot analysis of total cell extracts and mitochondria isolated from either the cerebral cortex or hippocampus of experimental animals was performed. In addition, Mfn2 was localized intracellularly by laser scanning confocal microscopy. It was demonstrated that 15‑min ischemia, or 15‑min ischemia followed by 1, 3, 24 or 72 h of reperfusion, was associated with a marked decrease of the Mfn2 protein in mitochondria isolated from the cerebral cortex, but not in hippocampal mitochondria. Moreover, a translocation of the Mfn2 protein to the cytoplasm was documented immediately after global brain ischemia in the neurons of the cerebral cortex by laser scanning confocal microscopy. Mfn2 translocation was followed by decreased expression of Mfn2 during reperfusion. Markedly elevated levels of the VDAC1 protein were also documented in total cell extracts isolated from the hippocampus of rats after 15 min of global brain ischemia followed by 3 h of reperfusion, and from the cerebral cortex of rats after 15 min of global brain ischemia followed by 72 h of reperfusion. The mitochondrial Mfn2 release observed during the early stages of reperfusion may thus represent an important mechanism of mitochondrial dysfunction associated with neuronal dysfunction or death induced by global brain ischemia.

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