MK-801 is cytotoxic to microglia in vitro and its cytotoxicity is attenuated by glutamate, other excitotoxic agents and atropine: Possible presence of glutamate receptor and muscarinic receptor on microglia

doi:10.1016/S0006-8993(01)02114-X

Brain Research

Volume 897, Issues 1–2, 6 April 2001, Pages 204-206

https://doi.org/10.1016/S0006-8993(01)02114-X Get rights and content

Abstract

We examined the cytotoxicity of MK-801 on cultured microglia and demonstrated its cytotoxicity. Cytotoxicity of MK-801 was reduced by the addition of l-glutamate, kainate and NMDA. The action of MK-801 was due to the direct action of microglia. It suggested the existence of glutamate receptor in microglia. Cytotoxicity of MK-801 was reduced by the addition of atropine sulfate which suggested the presence of muscarinic receptor in microglia.

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The role of non-synaptic extracellular glutamate
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Citation Excerpt :
Although kainate administration can induce microglial activation and tumor necrosis factor-α release (Zhu et al., 2010), the existence of kainate receptors in microglia has been questioned (Noda et al., 2000). The stimulation of NMDA receptors induces microglia activation in the brain (Acarin et al., 1996) and the inhibition of NMDA receptors (MK-801) decreases the in vitro toxicity of GLU (Hirayama and Kuriyama, 2001), however available evidence suggesting a microglial modulation by NMDA receptors needs to be contrasted (Streit et al., 1992; Thomas and Kuhn, 2005). Metabotropic GLU receptors have been identified in microglia (Biber et al., 1999), and their stimulation can induce microglia activation (Group I and Group II) (Farso et al., 2009; Kaushal and Schlichter, 2008), tumor necrosis factor-α release (Group II) (Kaushal and Schlichter, 2008), and adenilate cyclase inhibition (Group III) (Taylor et al., 2003).
Although there are some mechanisms which allow the direct crossing of substances between the cytoplasm of adjacent cells (gap junctions), most substances use the extracellular space to diffuse between brain cells. The present work reviews the behavior and functions of extracellular glutamate (GLU). There are two extracellular pools of glutamate (GLU) in the brain, a synaptic pool whose functions in the excitatory neurotransmission has been widely studied and an extrasynaptic GLU pool although less known nonetheless is gaining attention among a growing number of researchers. Evidence accumulated over the last years shows a number of mechanisms capable of releasing glial GLU to the extracellular medium, thus modulating neurons, microglia and oligodendrocytes, and regulating the immune response, cerebral blood flow, neuronal synchronization and other brain functions. This new scenario is expanding present knowledge regarding the role of GLU in the brain under different physiological and pathological conditions.
This article is part of a Special Issue entitled ‘Extrasynaptic ionotropic receptors’.
Combinations of ketamine and atropine are neuroprotective and reduce neuroinflammation after a toxic status epilepticus in mice
2012, Toxicology and Applied Pharmacology
Citation Excerpt :
This was tested in our paradigm with KET/AS treatments (see below). Although AS may contribute to the anti-inflammatory effect by acting on glial cells (Hirayama and Kuriyama, 2001), it did not prevent soman-induced brain inflammation as judged by the results obtained in our SOMAN CONTROL group. The mRNA levels of pro-inflammatory mediators detected in the SOMAN CONTROL group are in accordance to former studies (Dhote et al., 2007; Williams et al., 2003) showing an early and long-lasting mRNA induction of cytokines and adhesion molecules.
Epileptic seizures and status epilepticus (SE) induced by the poisoning with organophosphorus nerve agents (OP), like soman, are accompanied by neuroinflammation whose role in seizure-related brain damage (SRBD) is not clear. Antagonists of the NMDA glutamate ionotropic receptors are currently among the few compounds able to arrest seizures and provide neuroprotection even during refractory status epilepticus (RSE). Racemic ketamine (KET), in combination with atropine sulfate (AS), was previously shown to counteract seizures and SRBD in soman-poisoned guinea-pigs. In a mouse model of severe soman-induced SE, we assessed the potentials of KET/AS combinations as a treatment for SE/RSE-induced SRBD and neuroinflammation. When starting 30 min after soman challenge, a protocol involving six injections of a sub-anesthetic dose of KET (25 mg/kg) was evaluated on body weight loss, brain damage, and neuroinflammation whereas during RSE, anesthetic protocols were considered (KET 100 mg/kg). After confirming that during RSE, KET injection was to be repeated despite some iatrogenic deaths, we used these proof-of-concept protocols to study the changes in mRNA and related protein contents of some inflammatory cytokines, chemokines and adhesion molecules in cortex and hippocampus 48 h post-challenge. In both cases, the KET/AS combinations showed important neuroprotective effects, suppressed neutrophil granulocyte infiltration and partially suppressed glial activation. KET/AS could also reduce the increase in mRNA and related pro-inflammatory proteins provoked by the poisoning. In conclusion, the present study confirms that KET/AS treatment has a strong potential for SE/RSE management following OP poisoning. The mechanisms involved in the reduction of central neuroinflammation remain to be studied.
Activation of microglia by neuronal activity: Results from a new in vitro paradigm based on neuronal-silicon interfacing technology
2010, Brain, Behavior, and Immunity
Citation Excerpt :
In models of excitotoxic cell death, phagocytic microglia contribute to neuronal degeneration and cell loss (Chang et al., 2006; Cho et al., 2008; Kim et al., 2009). Since pilocarpine and kainic acid act at neurotransmitter receptors which microglia themselves can express (Hirayama and Kuriyama, 2001; Pocock and Kettenmann, 2007), it is difficult to separate their direct effects on microglia from the effects of surrounding neuronal and astrocytic activity. However, evidence from chemical seizure models is corroborated by the fact that electrically induced status epilepticus produces similar effects on microglia (Aronica et al., 2001; Bonde et al., 2006; Holtman et al., 2009).
Cognition and behavior primarily arise from the communication that occurs between brain cells. By using photoconductive stimulation to trigger localized regions of neuronal action potentials and astrocyte Ca²⁺ waves in dissociated rat hippocampal cultures, we can directly study microglia behavior in response to physiological and pathological levels of activity. Connections between neurons can be modified by microglia, which regulate gap junctions and synapses through secretion of proteins such as cytokines, proteases and neurotrophic factors. Activated microglia participate in bidirectional communication with the excitable tissues that they support. Through feedback from the many ion channels and surface receptors they express, microglia are informed of neuronal and astrocytic activity that may indicate disruption in the homeostasis of the CNS. Such disturbances alert microglia to locations of such activity and promote their transformation into a reactive state, in which they perform adaptive functions that can be either neuroprotective, neurotoxic, or neuromodulatory. Under physiological conditions, normal brain activity has the effect of suppressing microglia inflammatory responses. This report summarizes available data about the interaction of microglia and brain activity and presents a new in vitro paradigm to study the mechanisms involved. We propose that photoconductive stimulation is a powerful tool for studying the cellular and molecular mechanisms underlying the dynamic interactions between neurons, astrocytes and microglia.
Changes in neurotransmitter levels and proinflammatory cytokine mRNA expressions in the mice olfactory bulb following nanoparticle exposure
2008, Toxicology and Applied Pharmacology
Recently, there have been increasing reports that nano-sized component of particulate matter can reach the brain and may be associated with neurodegenerative diseases. Previously, our laboratory has studied the effect of intranasal instillation of nano-sized carbon black (CB) (14 nm and 95 nm) on brain cytokine and chemokine mRNA expressions and found that 14-nm CB increased IL-1β, TNF-α, CCL2 and CCL3 mRNA expressions in the olfactory bulb, not in the hippocampus of mice. To investigate the effect of a single administration of nanoparticles on neurotransmitters and proinflammatory cytokines in a mouse olfactory bulb, we performed in vivo microdialysis and real-time PCR methods. Ten-week-old male BALB/c mice were implanted with guide cannula in the right olfactory bulb and, 1 week later, were instilled vehicle or CB (14 nm, 250 μg) intranasally. Six hours after the nanoparticle instillation, the mice were intraperitoneally injected with normal saline or 50 μg of bacteria cell wall component lipoteichoic acid (LTA), which may potentiate CB-induced neurologic effect. Extracellular glutamate and glycine levels were significantly increased in the olfactory bulb of CB-instilled mice when compared with vehicle-instilled control mice. Moreover, we found that LTA further increased glutamate and glycine levels. However, no alteration of taurine and GABA levels was observed in the olfactory bulb of the same mice. We also detected immunological changes in the olfactory bulb 11 h after vehicle or CB instillation and found that IL-1β mRNA expression was significantly increased in CB- and LTA-treated mice when compared with control group. However, TNF-α mRNA expression was increased significantly in CB- and saline-treated mice when compared with control group. These findings suggest that nanoparticle CB may modulate the extracellular amino acid neurotransmitter levels and proinflammatory cytokine IL-1 β mRNA expressions synergistically with LTA in the mice olfactory bulb.
Memantine protects against LPS-induced neuroinflammation, restores behaviorally-induced gene expression and spatial learning in the rat
2006, Neuroscience
Citation Excerpt :
The microglia that remained in the LPS-infused animals treated with memantine presented a morphology characteristic of a less advanced activation state (Fig. 2D). It has been reported that the treatment with the selective NMDAR antagonist, MK-801, is cytotoxic to microglia in vitro (Hirayama and Kuriyama, 2001). Thus, the possibility that memantine may have reduced the number of activated microglia by reducing the total number of resident microglia was examined.
Neuroinflammation is reliably associated with the pathogenesis of a number of neurodegenerative diseases, and can be detected by the presence of activated microglia. Neuroinflammation can be induced by chronic lipopolysaccharide (LPS) infusion into the 4th ventricle of the rat resulting in region-selective microglia activation and impaired hippocampal-dependent memory. Furthermore, this treatment results in altered behaviorally-induced expression of the immediate early gene Arc, indicating altered network activity. LPS is known to activate microglia directly, leading to increased glutamate release, and in enhanced N-methyl-d-aspartate (NMDA) -dependent signaling. Taken together, the foregoing suggests that decreasing NMDA receptor activation during early stages of chronic neuroinflammation should reduce a) microglia activation, b) overexpression of Arc, and c) spatial memory deficits. Memantine, a low to moderate affinity open channel uncompetitive NMDA receptor antagonist, at low doses was used here to test these hypotheses. Rats were chronically infused into the 4th ventricle for 28 days with LPS alone, vehicle alone (via osmotic minipump) or LPS and memantine (10 mg/kg/day memantine s.c.). The results reported here demonstrate that memantine reduces OX6-immunolabeling for activated microglia, spares resident microglia, returns Arc (activity-regulated cytoskeletal associated protein, protein) -expressing neuronal populations to control levels (as revealed by Arc immunolabeling and fluorescence in situ hybridization), and ameliorates the spatial memory impairments produced by LPS alone. These data indicate that memantine therapy at low doses, recreating plasma levels similar to those of therapeutic doses in human, acts in part through its ability to reduce the effects of neuroinflammation, resulting in normal gene expression patterns and spatial learning.
Combined, these findings suggest that low, therapeutically relevant doses of memantine delivered early in the development of neuroinflammation-influenced diseases may confer neural and cognitive protection.
THE EFFECTS OF MEMANTINE AND MK801 ON NMDA RECEPTOR SWITCHING 2B AND 2A SUBUNITS IN HIPPOCAMPAL CELL CULTURE
2023, Archivos de Neurociencias

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Short communicationMK-801 is cytotoxic to microglia in vitro and its cytotoxicity is attenuated by glutamate, other excitotoxic agents and atropine: Possible presence of glutamate receptor and muscarinic receptor on microglia

Abstract

Brain Res.

Neuropharmacology

J. Immunol. Methods

Neurosci. Lett.

J. Neuroimmunol.

Brain Res.

Expression and signaling of group 1 metabotropic glutamate receptors in astrocytes and microglia

J. Neurochem.

Integrated evaluation of central nervous system lesions: stains for neurons, astrocytes, and microglia reveal the spatial and temporal features of MK-801-induced neuronal necrosis in the rat cerebral cortex

Toxicol. Pathol.

Short communication
MK-801 is cytotoxic to microglia in vitro and its cytotoxicity is attenuated by glutamate, other excitotoxic agents and atropine: Possible presence of glutamate receptor and muscarinic receptor on microglia