6-methyl-2-(phenylethynyl)pyridine and Hypoxia

Glioma is a primary brain tumor with high frequency and dismal prognosis. As there is no permanent cure available, identifying new therapy or mediator to augment the effectiveness of existing therapy is urgently needed. In the current study we tested the effect of group I metabotropic glutamate receptors (mGluRs): mGluR1 and mGluR5 on the viability of glioma cell lines. We analyzed cell viability using lactate dehydrogenase (LDH) release assay and evaluated apoptosis by propidium iodide (PI) staining. We used qPCR to evaluate change in mitochondrial gene expression and Western blot to evaluate the phosphorylation of Akt and ERK. Inhibition of mGluR5 by a selective antagonist MPEP under hypoxia promoted cell death, and induced expression of mitochondrial oxidative function related genes, with concurrent lowering of AKT phosphorylation level in glioma cell lines. Akt activation reversed mGluR5 inhibition on hypoxia-induced glioma cell death. These results suggest mGluR5 as a potential therapeutic target for hypoxic tumors such as malignant glioma.

Topics: Apoptosis; Brain Neoplasms; Cell Death; Cell Line, Tumor; Cell Survival; Excitatory Amino Acid Antagonists; Glioma; Humans; Hypoxia; Phosphorylation; Pyridines; Receptor, Metabotropic Glutamate 5; Signal Transduction

The influence of the selective antagonism of metabotropic glutamate receptor subtype 5 (mGluR5) by MPEP (2-methyl-6-(phenylethynyl)-pyridine) on some behaviors was tested in control groups of rats and in rats exposed to short-term hypoxia once or to repeated episodes of hypoxia.. We used the following methods: the open field test, the passive avoidance test and the object recognition test. Experimental hypoxia was produced by placing rats in a glass chamber flushed with a mixture of 2% O2 in N2.. MPEP applied intravenously (IV) at the dose of 1 mg kg-1 significantly enhanced locomotor and exploratory activity, impaired acquisition, but improved consolidation and retrieval in the passive avoidance situation and did not alter rats' activity in the object recognition test. The single short-term hypoxia significantly inhibited motility of rats and profoundly impaired acquisition, consolidation and retrieval processes, but the positive effect of MPEP on retrieval was preserved. Hypoxia also did not influence the activity of rats in the object recognition object. The repeated episodes of short-term hypoxia were induced for five consecutive days and it also inhibited motility of rats, but did not change consolidation and retrieval processes. The episodes of hypoxia significantly diminished the beneficial effect of MPEP on consolidation and retrieval, and also the enhancement of locomotor and exploratory activity. MPEP, used in rats subjected to the single or the repeated episodes of short-term hypoxia, did not change recognition memory.. MPEP used before the single episode of hypoxia only, had beneficial effect on retrieval.

Topics: Animals; Avoidance Learning; Behavior, Animal; Excitatory Amino Acid Antagonists; Exploratory Behavior; Hypoxia; Injections, Intravenous; Locomotion; Male; Memory; Pyridines; Rats; Rats, Wistar; Reaction Time; Receptor, Metabotropic Glutamate 5; Receptors, Metabotropic Glutamate; Recognition, Psychology; Time Factors

Group I metabotropic glutamate (mGlu) receptors (i.e. mGlu1 and mGlu5) coupled to phospholipase C have been widely investigated for their possible role in excitotoxic and post-ischemic neuronal death. Recently, phospholipase C has been shown to directly stimulate the activity of poly(ADP-ribose) polymerase (PARP), a nuclear enzyme involved in DNA repair that has been proposed to play a key role in necrotic cell death. In this study, we investigated whether the stimulation of group I mGlu receptors leads to an increase in PARP activity, as detected by flow cytometry, immunodot blot and immunocytochemistry, both in baby hamster kidney cells transfected with mGlu1a or mGlu5a receptors and in cultured cortical cells. Our results show that the group I mGlu receptor agonist DHPG elicited a significant increase in PARP activity that was completely abolished by the administration of the mGlu1 antagonist 3-MATIDA and partially prevented, in cortical neurons, by the mGlu5 antagonist MPEP. To evaluate whether this pathway is involved in post-ischemic neuronal death, we used a sublethal model of oxygen-glucose deprivation in mixed cortical cell cultures. DHPG exacerbated neuronal death, and this effect was significantly prevented by the application of the PARP inhibitor DPQ. This novel pathway may contribute to the effects of mGlu1 receptors in the mechanisms leading to post-ischemic neuronal death.

Topics: Animals; Animals, Newborn; Blotting, Western; Cells, Cultured; Cerebral Cortex; Cricetinae; Drug Interactions; Enzyme Activation; Enzyme Inhibitors; Excitatory Amino Acid Antagonists; Fluorescent Antibody Technique; Gene Expression Regulation, Enzymologic; Glial Fibrillary Acidic Protein; Glucose; Hydrogen Peroxide; Hypoxia; Isoquinolines; Methoxyhydroxyphenylglycol; Mice; Neuroglia; Neurons; Piperidines; Poly(ADP-ribose) Polymerases; Pyridines; Receptors, Metabotropic Glutamate; Thiophenes; Time Factors; Transfection