Astrocyte mGlu2/3-mediated cAMP potentiation is calcium sensitive: studies in murine neuronal and astrocyte cultures

Moldrich, Randall X; Apricó, Karina; Diwakarla, Shanti; O'Shea, Ross D; Beart, Philip M

doi:10.1016/s0028-3908(02)00111-9

Cited by 41 publications

(41 citation statements)

References 53 publications

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“…Cells from the cerebral neocortex were cultured from Swiss white embryonic mice (day 15 to 16) 19 and seeded at 0.12ϫ10 6 …”

Section: Neuronal Cell Culturesmentioning

confidence: 99%

“…Viability was alternatively quantified by the ability of surviving cells to reduce 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to formazan. 19 …”

Section: Cytotoxicity Studiesmentioning

confidence: 99%

“…19 Where indicated, tPA or DSPA (500 nmol/L) was added alone or in combination to cells 5 minutes before addition of 70 mol/L NMDA. Cellular fluorescence was measured in relative fluorescence units using the Fluoroskan Ascent fluorometer.…”

Section: Calcium Mobilization Studiesmentioning

confidence: 99%

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Vampire Bat Salivary Plasminogen Activator (Desmoteplase) Inhibits Tissue-Type Plasminogen Activator-Induced Potentiation of Excitotoxic Injury

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Moldrich

Beart

et al. 2005

Stroke

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Background and Purpose-In contrast to tissue-type plasminogen activator (tPA), vampire bat (Desmodus rotundus) salivary plasminogen activator (desmoteplase [DSPA]) does not promote excitotoxic injury when injected directly into the brain. We have compared the excitotoxic effects of intravenously delivered tPA and DSPA and determined whether DSPA can antagonize the neurotoxic and calcium enhancing effects of tPA. Methods-The brain striatal region of wild-type c57 Black 6 mice was stereotaxically injected with N-methyl-D-Aspartate (NMDA); 24 hour later, mice received an intravenous injection of tPA or DSPA (10 mg/kg) and lesion size was assessed after 24 hours. Cell death and calcium mobilization studies were performed using cultures of primary murine cortical neurons. Results-NMDA-mediated injury was increased after intravenous administration of tPA, whereas no additional toxicity was seen after administration of DSPA. Unlike DSPA, tPA enhanced NMDA-induced cell death and the NMDAmediated increase in intracellular calcium levels in vitro. Moreover, the enhancing effects of tPA were blocked by DSPA. Conclusions-Intravenous administration of tPA promotes excitotoxic injury, raising the possibility that leakage of tPA from the vasculature into the parenchyma contributes to brain damage. The lack of such toxicity by DSPA further encourages its use as a thrombolytic agent in the treatment of ischemic stroke.

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“…Cells from the cerebral neocortex were cultured from Swiss white embryonic mice (day 15 to 16) 19 and seeded at 0.12ϫ10 6 …”

Section: Neuronal Cell Culturesmentioning

confidence: 99%

“…Viability was alternatively quantified by the ability of surviving cells to reduce 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to formazan. 19 …”

Section: Cytotoxicity Studiesmentioning

confidence: 99%

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Vampire Bat Salivary Plasminogen Activator (Desmoteplase) Inhibits Tissue-Type Plasminogen Activator-Induced Potentiation of Excitotoxic Injury

Reddrop

Moldrich

Beart

et al. 2005

Stroke

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“…Cells were visualised under bright field microscopy and TUNELpositive staining was observed only in neurones with shrunken, pyknotic morphology. Intracellular calcium studies Intracellular calcium was determined by using Fluo-3AM calcium-binding dye (Molecular Probes) as described in detail by Moldrich et al (2002). Briefly, cells were loaded with 10 μM Fluo-3AM in HEPES-buffered saline (HBS).…”

Section: Drug Exposure and Cellular Viability Assaysmentioning

confidence: 99%

Establishment of primary cultures of rat olfactory bulb under serum-free conditions for studies of cellular injury

2005

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Olfactory dysfunction has been implicated in various neurodegenerative diseases including Parkinson's and Alzheimer's disease but, despite intense interest in the neurobiology of the olfactory bulb (OB), studies of neurodegenerative mechanisms have not been attempted in primary OB cultures. This study was aimed at developing a primary OB culture under serum-free conditions in order to investigate injury and excitotoxicity in vitro. Olfactory bulbs from rat pups were rapidly trypsinised and mechanically dissociated and the resultant single cell suspension was centrifuged through a high bovine serum albumin concentration gradient to reduce cellular debris before being seeded in multi-well culture plates. Cells were plated in neurobasal medium containing 0.5 mM glutamine, 25 mM K(+), 2% B27 and 10% fetal calf serum (FCS) for 24 h and, after 1 day in vitro (div1), were maintained without FCS. At div8, neurones exhibited extensive neuritic networks, were present as a monolayer and were mainly bipolar and immunopositive for gamma-aminobutyric acid indicating that they were intrinsic OB neurones. At div8, neurones (positive for microtubule-associated protein-2, 73%) predominated over astrocytes (positive for glial fibrillary acidic protein, 27%). Cellular injury produced by staurosporine, hydrogen peroxide and kainate, when assessed by morphological and biochemical procedures, was shown to be concentration-dependent and significantly reduced the numbers of neurones and astrocytes. Further analyses of kainate-induced injury revealed the presence of TUNEL-positive cells (indicative of apoptosis) and increases in intracellular free calcium, both of which were antagonised by CNQX. Thus, the serum-free culture developed here is amenable to morphological and high throughput neurochemical analyses of mechanisms contributing to the injury of OB neurones in vitro.

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“…The group II mGlu receptors function primarily as autoreceptors modulating GABAergic and monoaminergic neurotransmitter release directly (Cartmell and Schoepp, 2000) and via various modulatory actions on astrocytic mGlu3 receptors (Bruno et al, 1998;Moldrich et al, 2002). They also play a major role in modulating the release of L-glutamate, and they are highly expressed in the hippocampus, amygdala, and prefrontal cortex (Ohishi et al, 1993a,b;Schoepp et al, 2003).…”

mentioning

confidence: 99%

Pharmacological and Pharmacokinetic Properties of a Structurally Novel, Potent, and Selective Metabotropic Glutamate 2/3 Receptor Agonist: In Vitro Characterization of Agonist (–)-(1R,4S,5S,6S)-4-Amino-2-sulfonylbicyclo[3.1.0]-hexane-4,6-dicarboxylic Acid (LY404039)

Rorick-Kehn

Johnson²,

Burkey³

et al. 2007

J Pharmacol Exp Ther

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Group II metabotropic glutamate (mGlu) receptor agonists, including (1S,2S,5R,6S)-2-aminobicyclo[3.1.0]hexane-2,6-dicarboxylate monohydrate (LY354740) and (Ϫ)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate (LY379268), have demonstrated efficacy in animal models of anxiety and schizophrenia, and LY354740 decreased anxiety in human subjects. Herein, we report the in vitro pharmacological profile and pharmacokinetic properties of another potent, selective, and structurally novel mGlu2/3 receptor agonist, (Ϫ)-(1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid (LY404039) and provide comparisons with LY354740. Similar to LY354740, LY404039 is a nanomolar potent agonist at recombinant human mGlu2 and mGlu3 receptors (K i ϭ 149 and 92, respectively) and in rat neurons expressing native mGlu2/3 receptors (K i ϭ 88). LY404039 is highly selective for mGlu2/3 receptors, showing more than 100-fold selectivity for these receptors, versus ionotropic glutamate receptors, glutamate transporters, and other receptors targeted by known anxiolytic and antipsychotic medications. Functionally, LY404039 potently inhibited forskolin-stimulated cAMP formation in cells expressing human mGlu2 and mGlu3 receptors. Electrophysiological studies indicated that LY404039 suppressed electrically evoked excitatory activity in the striatum, and serotonininduced L-glutamate release in the prefrontal cortex; effects reversed by LY341495. These characteristics suggest LY404039 modulates glutamatergic activity in limbic and forebrain areas relevant to psychiatric disorders; and that, similar to LY354740, it works through a mechanism that may be devoid of negative side effects associated with current antipsychotics and anxiolytics. Interestingly, despite the slightly lower potency (ϳ2-5-fold) of LY404039 versus LY354740 in binding, functional, and electrophysiological assays, LY404039 demonstrated higher plasma exposure and better oral bioavailability in pharmacokinetic experiments. Collectively, the current data indicate that LY404039 may be valuable in the treatment of neuropsychiatric disorders, including anxiety and psychosis.Glutamate receptors are made up of two receptor families: ionotropic glutamate (iGlu) and metabotropic glutamate (mGlu) receptors. iGlu receptors are ligand-gated ion channels that mediate fast synaptic transmission and include NMDA, ␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, and kainate receptor subtypes. Metabotropic glutamate receptors are G protein-linked receptors that mediate multiple second messenger systems (Pin and Duvoisin, 1995; Cart-Article, publication date, and citation information can be found at

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Astrocyte mGlu2/3-mediated cAMP potentiation is calcium sensitive: studies in murine neuronal and astrocyte cultures

Cited by 41 publications

References 53 publications

Vampire Bat Salivary Plasminogen Activator (Desmoteplase) Inhibits Tissue-Type Plasminogen Activator-Induced Potentiation of Excitotoxic Injury

Vampire Bat Salivary Plasminogen Activator (Desmoteplase) Inhibits Tissue-Type Plasminogen Activator-Induced Potentiation of Excitotoxic Injury

Establishment of primary cultures of rat olfactory bulb under serum-free conditions for studies of cellular injury

Pharmacological and Pharmacokinetic Properties of a Structurally Novel, Potent, and Selective Metabotropic Glutamate 2/3 Receptor Agonist: In Vitro Characterization of Agonist (–)-(1R,4S,5S,6S)-4-Amino-2-sulfonylbicyclo[3.1.0]-hexane-4,6-dicarboxylic Acid (LY404039)

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