Riluzole promotes survival of rat motoneurons in vitro by stimulating trophic activity produced by spinal astrocyte monolayers (original) (raw)
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Effect of prolonged riluzole exposure on cultured motoneurons in a mouse model of ALS
Journal of Neurophysiology, 2012
Riluzole is the only FDA-approved drug to treat amyotrophic lateral sclerosis, but its long-term effects on motoneurons are unknown. Therefore, we treated primary mouse spinal cord cultures with 2 μM riluzole for 4–9 days and then used whole cell patch clamp to record the passive and active properties of both wild-type and SOD1G93A motoneurons. At this concentration, riluzole blocks >50% of the sodium component of a persistent inward current that plays a major role in determining motoneuron excitability. Prolonged riluzole treatment significantly decreased the amplitude of the persistent inward current. This effect was specific for SOD1G93A motoneurons, where the amplitude decreased by 55.4%. In addition, prolonged treatment hyperpolarized the resting membrane potential as well as the voltage onset and voltage maximum of the persistent inward current (∼2–3 mV in each case). These effects appeared to offset one another and resulted in no change in the firing properties. In a subse...
Neuroscience, 2011
Kainate-mediated excitotoxicity of organotypic spinal cord cultures is an in vitro model advantageous to investigate basic mechanisms of acute spinal injury and its pharmacological neuroprotection. Using such cultures, the putative neuroprotective agent riluzole applied at 5 M (plasma therapeutic concentration) was studied for its ability to prevent neurotoxicity evoked by 1 h administration of kainate. We monitored real-time release of glutamate, release of lactate dehydrogenase (LDH) (cell damage marker), occurrence of cell pyknosis, the number of surviving neurons and motoneurons, and cell culture metabolic activity. Co-applied riluzole strongly blocked the kainate-evoked early rise in extracellular glutamate (via calcium dependent or independent processes) and suppressed LDH release (limited to <20% of total). Although there were no significant cell losses within the first h after kainate washout, pyknosis, fewer neurons and motoneurons were observed 24 h later. MTT assay demonstrated that surviving cells were metabolically competent. Co-application of kainate and tetrodotoxin also failed to protect spinal cord slices 24 h later. When riluzole application begun at kainate washout and continued for 24 h, significant neuroprotection was observed for neurons in the central and dorsal regions, while ventral horn cells (including motoneurons) were not protected. Our data suggest that riluzole neuroprotection against excitotoxicity was feasible, although it paradoxically required delayed drug administration, and was not extended to the ventral horn. We propose that riluzole was acting on yet-unidentified processes downstream of glutamate release and receptor activation. Deciphering their identity and role in cell death mechanisms may be an important goal to develop neuroprotection.
Riluzole promotes cell survival and neurite outgrowth in rat sensory neurones in vitro
European Journal of Neuroscience, 2006
This study explored the effects of riluzole administration on cell survival and neurite growth in adult and neonatal rat dorsal root ganglion (DRG) neurones in vitro. Neuronal survival was assessed by comparing numbers of remaining neurones in vehicle-and riluzole-treated cultures. A single dose of 0.1 lm riluzole was sufficient to promote neuronal survival in neonatal DRG cultures, whereas repeated riluzole administration was necessary in adult cultures. However, a single administration of riluzole was sufficient to induce neuritogenesis, promote neurite branching and enhance neurite outgrowth in both neonatal and adult DRG cultures. The effects of a single dose of riluzole on adult DRG neurones after peripheral nerve or dorsal root injury were also studied in vitro at 48 h. For both types of injury, riluzole enhanced neurite outgrowth in terms of number, length and branch pattern significantly more on the injured side as compared with the contralateral side. No effect was seen on cell survival. The results suggest that, in addition to its cell survival effects, riluzole has novel growth-promoting effects on sensory neurones in vitro and that riluzole may offer a new way to promote sensory afferent regeneration following peripheral injury.
Neuroscience, 2010
Riluzole is clinically approved for the treatment of motoneuron disease. We have previously shown that this drug is neuroprotective for both sensory neurons and motoneurons and promotes neurite outgrowth [Bergerot A, Shortland PJ, Anand P, Hunt SP, Carlstedt T (2004) Exp Neurol 187(2):359 -366; Shortland PJ, Leinster VH, White W, Robson LG (2006) Eur J Neurosci 24:3343-3353]. This study explored the effects of exogenous administration of 0.1 M riluzole on the neurite growth of specific subpopulations of adult rat dorsal root ganglion (DRG) neurons in vitro. Neuronal branching and neurite length were measured in calcitonin gene related peptide (CGRP), Griffonia simplicifolia Isolectin B4 (IB4), N52 and parvalbumin positive neuronal subpopulations. Riluzole was found to enhance neurite branching in both CGRP and IB4 positive neurons compared to vehicle treated cultures. However, neurite length was only significantly increased in CGRP positive neurons in riluzole treated cultures.
Experimental Neurology, 2006
The wobbler mouse is one of the most useful models of motoneuron degeneration, characterized by selective motoneuronal death in the cervical spinal cord. We carried out two parallel studies in wobbler mice, comparing the anti-glutamatergic drug riluzole and the AMPA receptor antagonist RPR119990. Mice were treated with 40 mg/kg/day of riluzole or with 3 mg/kg/day of RPR119990 from the 4th to the 12th week of age. Here, we show that chronic treatment with riluzole improves motor behavior, prevents biceps muscle atrophy and decreases the amount of motoneuron loss in treated wobbler mice. Chronic treatment with the AMPA antagonist RPR119990 is ineffective in improving motor impairment, in reducing motoneuronal loss and muscular atrophy in treated mice. These results, together with the unchanged immunostaining for the AMPA receptor subunit GluR2 in wobbler mice, suggest that AMPA receptor-mediated injury is unlikely to be involved in neurodegeneration in wobbler disease, and that the protective effect of riluzole in wobbler mice seems to be independent of its anti-glutamatergic activity, as suggested in other models of neurodegeneration. Immunostaining of cervical spinal cord sections shows that in riluzole-treated wobbler mice BDNF expression is significantly increased in motoneurons with no changes in the high-affinity receptor Trk-B. Our data confirm that riluzole has beneficial effects in wobbler mice, and suggest that these effects could be associated to the increased levels of the neurotrophic and neuroprotective factor BDNF.
Chronic inhibitory effect of riluzole on trophic factor production
Experimental neurology, 2015
Riluzole is the only FDA approved drug for the treatment of amyotrophic lateral sclerosis (ALS). However, the drug affords moderate protection to ALS patients, extending life for a few months by a mechanism that remains controversial. In the presence of riluzole, astrocytes increase the production of factors protective to motor neurons. The stimulation of trophic factor production by motor neuron associated cells may contribute to riluzole's protective effect in ALS. Here, we investigated the effects of media conditioned by astrocytes and Schwann cells acutely or chronically incubated with riluzole on trophic factor-deprived motor neuron survival. While acute riluzole incubation induced CT-1 secretion by astrocytes and Schwann cells, chronic treatment stimulated a significant decrease in trophic factor production compared to untreated cultures. Accordingly, conditioned media from astrocytes and Schwann cells acutely treated with riluzole protected motor neurons from trophic fact...
Experimental inhibition of reactive gliotic-like changes in astrocytic cultures
Experimental Neurology, 1988
Dibutylcyclic AMP (DiBucAMP) can induce astroblast-containing cultures to form cells which resemble reactive astrocytes observed in vivo. In the present study, myelin-or axolemmal-enriched fractions were assessed for the ability to inhibit (Di-BucAMP)-stimulated reactive-like changes in astrocytic cultures. Addition of exogenous myelin-or axolemmal-enriched fractions to DiBcAMPexposed cultures prevented drug-induced elevation of lactic dehydrogenase (LDH) 2 days after initial addition of the drug and moderated DiBucAMP-induced decreases in the enzyme's activity normally observed 5 days later. The proportion of reactive colonies (i.e., those in which more than 50% of cells are stellate-shape) was significantly lower in cultures exposed to the drug plus myelin or axolemma vs those exposed to DiBucAMP alone. The inhibitory factors in axolemmal fractions were heat sensitive (at IOOC), whereas those in myelin were not. Both fractions were inactivated by trypsin. Whole brain homogenates had no effect on diBucAMP-stimulated changes in culture. o 1988 Academic Press. Inc.
Neurochemistry International, 1994
Basic fibroblast growth factor (bFGF) increases neuronal survival and growth in cell cultures and stimulates functional recovery from brain lesion. In addition, bFGF is able to induce glial cell proliferation and differentiation. Recently, L-deprenyl has been shown to potentiate astrocyte reaction to a mechanical lesion and to possess a trophic-like activity in several experimental models. In the present paper, we have therefore investigated if the enhancing effect of L-deprenyl on astrocyte reactivity is accompanied by increased levels of bFGF. The effect of L-deprenyl (0.25 mg/kg/day) on bFGF immunoreactivity (IR) after the insertion of an injection cannula in rat neostriatum have been investigated. It has been found that subchronic L-deprenyl treatment potentiates both the lesion-induced increase of glial fibrillary acidic protein (GFAP) and bFGF IRs (P < 0.01). These data suggest that a possible mechanism for L-deprenyl-induced neuroprotection may be the activation of astrocytes associated with increased secretion of trophic factors that promote neuronal survival and growth. This "astrocyte-kinetic" action of L-deprenyl could represent a new therapeutical approach to increase trophic support of lesioned neurons.
Riluzole elevates GLT-1 activity and levels in striatal astrocytes
Neurochemistry International, 2012
Drugs which upregulate astrocyte glutamate transport may be useful neuroprotective compounds by preventing excitotoxicity. We set up a new system to identify potential neuroprotective drugs which act through GLT-1. Primary mouse striatal astrocytes grown in the presence of the growthfactor supplement G5 express high levels of the functional glutamate transporter, GLT-1 (also known as EAAT2) as assessed by Western blotting and 3 H-glutamate uptake assay, and levels decline following growth factor withdrawal. The GLT-1 transcriptional enhancer dexamethasone (0.1 or 1 μM) was able to prevent loss of GLT-1 levels and activity following growth factor withdrawal. In contrast, ceftriaxone, a compound previously reported to enhance GLT-1 expression, failed to regulate GLT-1 in this system. The neuroprotective compound riluzole (100 μM) upregulated GLT-1 levels and activity, through a mechanism that was not dependent on blockade of voltage-sensitive ion channels, since zonasimide (1 mM) did not regulate GLT-1. Finally, CDP-choline (10 μM -1 mM), a compound which promotes association of GLT-1/ EAAT2 with lipid rafts was unable to prevent GLT-1 loss under these conditions. This observation extends the known pharmacological actions of riluzole, and suggests that this compound may exert its neuroprotective effects through an astrocyte-dependent mechanism.