Neurophobia Among Medical Students: Is Virtual Teaching the Answer?

OBJECTIVE: Neurophobia is well recognized as dissuading medical students from neurocentric specialties and limiting the success of neurology and neurosurgery teaching at medical school. Past studies have associated neurophobia with deficiencies in medical education. We performed a cross-sectional analysis of medical students' confidence and perceived level of knowledge in recognizing the following neurosurgical and neurological emergencies: ischemic stroke, hemorrhagic stroke, status epilepticus, subarachnoid hemorrhage, increased intracranial pressure, acute hydrocephalus, spinal cord injury, cauda equina syndrome, and traumatic brain injury. In addition, we assessed the usefulness of virtual seminars in neurosurgery and neurology teaching. METHODS: Medical students from King's College London were invited to a virtual teaching session. We obtained preteaching and postteaching scores for students' subjective ability to recognize specific neurologic and neurosurgical emergencies, along with their confidence in the subject. RESULTS: Ninety-seven medical students attended the teaching session. For our sample group's subjective rating on their confidence in neurology or neurosurgery as a subject, we obtained a mean score of 3.87 and a median score of 4. Across all domains, there was a significant forward shift in the distribution curve of scores after teaching. We obtained statistically significant differences for all 9 neurologic and neurosurgical emergencies evaluated in our questionnaire (asymptotic significance <0.001). Median scores for all 9 conditions improved after the teaching session, with >50% positive ranks seen within each group. Across the teaching modalities compared, placement teaching was the highest scoring, whereas online lectures received a better rating than in-person lectures. CONCLUSIONS: In neurosurgery teaching, virtual seminars may compensate for deficiencies that exist within medical education, hence limiting the effects of neurophobia.

Five Breakthroughs: A First Approximation of Brain Evolution From Early Bilaterians to Humans.

Retracing the evolutionary steps by which human brains evolved can offer insights into the underlying mechanisms of human brain function as well as the phylogenetic origin of various features of human behavior. To this end, this article presents a model for interpreting the physical and behavioral modifications throughout major milestones in human brain evolution. This model introduces the concept of a "breakthrough" as a useful tool for interpreting suites of brain modifications and the various adaptive behaviors these modifications enabled. This offers a unique view into the ordered steps by which human brains evolved and suggests several unique hypotheses on the mechanisms of human brain function.

What Behavioral Abilities Emerged at Key Milestones in Human Brain Evolution? 13 Hypotheses on the 600-Million-Year Phylogenetic History of Human Intelligence.

This paper presents 13 hypotheses regarding the specific behavioral abilities that emerged at key milestones during the 600-million-year phylogenetic history from early bilaterians to extant humans. The behavioral, intellectual, and cognitive faculties of humans are complex and varied: we have abilities as diverse as map-based navigation, theory of mind, counterfactual learning, episodic memory, and language. But these faculties, which emerge from the complex human brain, are likely to have evolved from simpler prototypes in the simpler brains of our ancestors. Understanding the order in which behavioral abilities evolved can shed light on how and why our brains evolved. To propose these hypotheses, I review the available data from comparative psychology and evolutionary neuroscience.

The Thalamus as a Blackboard for Perception and Planning.

It has been suggested that the thalamus acts as a blackboard, on which the computations of different cortical modules are composed, coordinated, and integrated. This article asks what blackboard role the thalamus might play, and whether that role is consistent with the neuroanatomy of the thalamus. It does so in a context of Bayesian belief updating, expressed as a Free Energy Principle. We suggest that the thalamus-as-a-blackboard offers important questions for research in spatial cognition. Several prominent features of the thalamus-including its lack of olfactory relay function, its lack of internal excitatory connections, its regular and conserved shape, its inhibitory interneurons, triadic synapses, and diffuse cortical connectivity-are consistent with a blackboard role.Different thalamic nuclei may play different blackboard roles: (1) the Pulvinar, through its reciprocal connections to posterior cortical regions, coordinates perceptual inference about "what is where" from multi-sense-data. (2) The Mediodorsal (MD) nucleus, through its connections to the prefrontal cortex, and the other thalamic nuclei linked to the motor cortex, uses the same generative model for planning and learning novel spatial movements. (3) The paraventricular nucleus may compute risk-reward trade-offs. We also propose that as any new movement is practiced a few times, cortico-thalamocortical (CTC) links entrain the corresponding cortico-cortical links, through a process akin to supervised learning. Subsequently, the movement becomes a fast unconscious habit, not requiring the MD nucleus or other thalamic nuclei, and bypassing the thalamic bottleneck.

An Attempt at a Unified Theory of the Neocortical Microcircuit in Sensory Cortex.

The neocortex performs a wide range of functions, including working memory, sensory perception, and motor planning. Despite this diversity in function, evidence suggests that the neocortex is made up of repeating subunits ("macrocolumns"), each of which is largely identical in circuitry. As such, the specific computations performed by these macrocolumns are of great interest to neuroscientists and AI researchers. Leading theories of this microcircuit include models of predictive coding, hierarchical temporal memory (HTM), and Adaptive Resonance Theory (ART). However, these models have not yet explained: (1) how microcircuits learn sequences input with delay (i.e., working memory); (2) how networks of columns coordinate processing on precise timescales; or (3) how top-down attention modulates sensory processing. I provide a theory of the neocortical microcircuit that extends prior models in all three ways. Additionally, this theory provides a novel working memory circuit that extends prior models to support simultaneous multi-item storage without disrupting ongoing sensory processing. I then use this theory to explain the functional origin of a diverse set of experimental findings, such as cortical oscillations.

An Uncertainty Visual Analytics Framework for fMRI Functional Connectivity.

Analysis and interpretation of functional magnetic resonance imaging (fMRI) has been used to characterise many neuronal diseases, such as schizophrenia, bipolar disorder and Alzheimer's disease. Functional connectivity networks (FCNs) are widely used because they greatly reduce the amount of data that needs to be interpreted and they provide a common network structure that can be directly compared. However, FCNs contain a range of data uncertainties stemming from inherent limitations, e.g. during acquisition, as well as the loss of voxel-level data, and the use of thresholding in data abstraction. Additionally, human uncertainties arise during interpretation due to the complexity in understanding the data. While existing FCN visual analytics tools have begun to mitigate the human ambiguities, reducing the impact of data limitations is an open problem. In this paper, we propose a novel visual analytics framework with three linked, purpose-designed components to evoke deeper interpretation of the fMRI data: (i) an enhanced FCN abstraction; (ii) a temporal signal viewer; and (iii) the anatomical context. Each component has been specifically designed with novel visual cues and interaction to expose the impact of uncertainties on the data. We augment this with two methods designed for comparing subjects, by using a small multiples and a marker approach. We demonstrate the enhancements enabled by our framework on three case studies of common research scenarios, using clinical schizophrenia data, which highlight the value in interpreting fMRI FCN data with an awareness of the uncertainties. Finally, we discuss our framework in the context of fMRI visual analytics and the extensibility of our approach.

White matter integrity alterations in post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) is a debilitating condition which can develop after exposure to traumatic stressors. Seventy-five adults were recruited from the community, 25 diagnosed with PTSD along with 25 healthy and 25 trauma-exposed age- and gender-matched controls. Participants underwent clinical assessment and magnetic resonance imaging. A previous voxel based morphometry (VBM) study using the same subject cohort identified decreased grey matter (GM) volumes within frontal/subcortical brain regions including the hippocampus, amygdala, and anterior cingulate cortex (ACC). This study examines the microstructural integrity of white matter (WM) tracts connecting the aforementioned regions/structures. Using diffusion tensor imaging, we investigated the integrity of frontal/subcortical WM tracts between all three subject groups. Trauma exposed subjects with and without PTSD diagnosis were identified to have significant disruption in WM integrity as indexed by decreased fractional anisotropy (FA) in the uncinate fasciculus (UF), cingulum cingulate gyrus (CCG), and corpus callosum (CC), when compared with healthy non-trauma-exposed controls. Significant negative correlations were found between total Clinician Administered PTSD scale (CAPS) lifetime clinical subscores and FA values of PTSD subjects in the right UF, CCG, CC body, and right superior longitudinal fasciculus (SLF). An analysis between UF and SLF FA values and VBM determined rostral ACC GM values found a negative correlation in PTSD subjects. Findings suggest that compromised WM integrity in important tracts connecting limbic structures such as the amygdala to frontal regions including the ACC (i.e., the UF and CCG) may contribute to impairments in threat/fear processing associated with PTSD.

Fiber pathway pathology, synapse loss and decline of cortical function in schizophrenia.

A quantitative cortical model is developed, based on both computational and simulation approaches, which relates measured changes in cortical activity of gray matter with changes in the integrity of longitudinal fiber pathways. The model consists of modules of up to 5,000 neurons each, 80% excitatory and 20% inhibitory, with these having different degrees of synaptic connectiveness both within a module as well as between modules. It is shown that if the inter-modular synaptic connections are reduced to zero while maintaining the intra-modular synaptic connections constant, then activity in the modules is reduced by about 50%. This agrees with experimental observations in which cortical electrical activity in a region of interest, measured using the rate of oxidative glucose metabolism (CMRglc(ox)), is reduced by about 50% when the cortical region is isolated, either by surgical means or by transient cold block. There is also a 50% decrease in measured cortical activity following inactivation of the nucleus of Meynert and the intra-laminar nuclei of the thalamus, which arise either following appropriate lesions or in sleep. This occurs in the model if the inter-modular synaptic connections require input from these nuclei in order to function. In schizophrenia there is a 24% decrease in functional anisotropy of longitudinal fasciculi accompanied by a 7% decrease in cortical activity (CMRglc(ox)).The cortical model predicts this, namely for a 24% decrease in the functioning of the inter-modular connections, either through the complete loss of 24% of axons subserving the connections or due to such a decrease in the efficacy of all the inter-modular connections, there will be about a 7% decrease in the activity of the modules. This work suggests that deterioration of longitudinal fasciculi in schizophrenia explains the loss of activity in the gray matter.

A model of neuregulin control of NMDA receptors on synaptic spines.

Neuregulin (Nrg) through its receptor ErbB4 modulates the activity of the N-Methyl-D-Aspartate (NMDA) receptor (NMDAR) at synapses. As modification of this pathway has been implicated in schizophrenia, it is of great interest to define it in precise quantitative terms. Kinetic models of the epidermal growth factor (EGF)/ErbB receptor signalling pathway describing activation, desensitization, and tyrosine phosphorylation of EGFR/ErbB followed by binding and activation of Src family kinases that is subsequently followed by phosphorylation of target proteins are available. We have adapted these to give a kinetic description of NMDAR modulation by Nrg that recapitulates the observed kinetics of autophosphorylation of the ErbB dimer as well as the modulation of the NMDAR by Src kinase, according to whether the kinases are activated or deactivated. This quantitative description of the Nrg/NMDAR pathway provides a model for experimental elucidation of what goes awry in animal models of schizophrenia.

A model of NMDA receptor control of F-actin treadmilling in synaptic spines and their growth.

Synaptic spines grow as a consequence of the formation of F-actin filaments at the spine head. The dynamics of F-actin in the spine head upon excitation of N-methy-D-aspartate (NMDA) receptors has recently been investigated experimentally, but there is no quantitative account of how these dynamic changes occur upon activation of these receptors; this we now supply. Dynamics of F-actin at the apex of lamellipodia have been investigated in detail, giving rise to the treadmilling theory of F-actin dynamics, involving catalysis by profilin, for which quantitative models are now available. Here, we adapt such a model to describe the dynamics of F-actin in the synaptic-spine head and show that it gives quantitative descriptions of this treadmilling phenomena which are well fitted by Monte Carlo simulations. Next, the means by which excitation of NMDA receptors enhances the activity of profilin through activity of the Rho small GTPase RhoA and the specific kinase ROCK is discussed. This is then used to model the NMDA receptor excitatory enhancement of profilin and so the treadmilling process of F-actin dynamics in spine growth. Such modelling provides a quantitative description of the synaptic-spine dynamics of the filamentous to globular actin ratio that is observed experimentally.

P2X7 regenerative-loop potentiation of glutamate synaptic transmission by microglia and astrocytes.

P2X7 purinergic receptors have been implicated in chronic neuropathic and neuroinflammatory pain as well as in depression. These receptors are predominantly found in the central nervous system on microglial cells and on glutamatergic nerve terminals. Here, we develop hypotheses concerning mechanisms by which transient high-frequency impulse firing in glutamatergic terminals, such as occurs in nociceptor terminals accompanying neuropathic/neuroinflammatory pain, can lead to long-lasting changes in neural network function that is mediated by surrounding glial cells. The hypothesis consists of two parts. In the first, glutamate released by low-frequency (2Hz) terminal action potentials is insufficient to generate postsynaptic action potentials, but these are generated by brief high-frequency input bursts. Glutamate released by these bursts is partly removed by transporters on the enveloping astrocyte processes and also excites AMPA receptors on these processes, which then release ATP. This ATP is partly metabolised to adenosine, which acts on presynaptic A1 receptors to inhibit glutamate release. The remaining ATP acts on the presynaptic P2X7 receptors to facilitate glutamate release by both the high-frequency burst of action potentials as well as by a continuous low-frequency (2Hz) action potential firing that occurs in the absence of a neuropathic/neuroinflammatory insult. The positive feedback of terminal glutamate release, triggering astrocyte ATP release and leading to further glutamate release through activation of P2X7 receptors, is then sufficient to allow the normal low-frequency (2Hz) action potentials to now elicit postsynaptic action potentials after the insult is removed. In the second part of this model, the high concentration of ATP derived from astrocytes at the terminal attracts microglia by chemotaxis. The P2X7 receptors on these microglia are then engaged, resulting in microglia secreting the cytokine TNFalpha. This acts on postsynaptic TNF-R1 receptors to increase the number of AMPA receptors there, thus enhancing the efficacy of synaptic transmission. The TNFalpha also acts on presynaptic TNF-R1 to increase the amount of glutamate released by each nerve terminal impulse. Experimental tests can be made of this hypothesis that P2X7 receptors on the presynaptic terminal and those on the microglia synergistically act to ensure feedback pathways that reset to a high level the efficacy of synaptic transmission, thus ensuring chronic neuropathic/neuroinflammatory pain even when the initial insult has subsided.

Glutamate induces directed chemotaxis of microglia.

Microglia in the brain possess dynamic processes that continually sample the surrounding parenchyma and respond to local insults by rapidly converging on the site of an injury. One of the chemotaxic agents responsible for this response is ATP. Here we show that the transmitter glutamate is another such chemotaxic agent. Microglia exposed to glutamate increase their cell membrane ruffling and migrate to a source of glutamate in cell culture and in spinal cord slices. This chemotaxis is meditated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and metabotropic glutamate receptors on the microglia. Chemotaxis is dependent on redistribution of actin filaments in the cells and on tubulin following receptor activation. Thus glutamate, which is released at synapses as well as from damaged cells, can mediate rapid chemotaxic responses from microglial cells.

A quantitative model of cortical spreading depression due to purinergic and gap-junction transmission in astrocyte networks.

Spreading depression (SD), a propagating wave of electrical silence in the cortex and archicortex, involves depolarization of neurons and astrocytes for approximately 1 min, due principally to a large increase in extracellular K+. SD is accompanied by large increases in extracellular ATP and is blocked by glutamate N-methyl-D-aspartate receptor antagonists. As a principal means of transmission between astrocytes is through their release of ATP, we have investigated if a model in which SD is driven by the effects of astrocyte waves of ATP interacting with waves of glutamate release from neurons and astrocytes can give a quantitative account of experimental observations on SD. We show that the characteristics of SD and the accompanying extracellular ionic changes can be accommodated by such a model-whether astrocyte transmission is principally through the release of ATP, as in archicortex (hippocampus) and spinal cord, or via gap junctions, as in the neocortex. Furthermore, these models give quantitative accounts of the effects on the characteristics of SD of agents toxic for astrocytes and of gap-junction blockers. Finally, an additional series of critical tests of the model is suggested.

Purinergic junctional transmission and propagation of calcium waves in cultured spinal cord microglial networks.

In order to elucidate the mechanisms of purinergic transmission of calcium (Ca(2 + )) waves between microglial cells, we have employed micro-photolithographic methods to form discrete patterns of microglia that allow quantitative measurements of Ca(2 + ) wave propagation. Microglia were confined to lanes 20-100 [Formula: see text] wide and Ca(2 + ) waves propagated from a point of mechanical stimulation, with a diminution in amplitude, for about 120 [Formula: see text]. The number of cells participating in propagation also decreased over this distance. Ca(2 + ) waves could propagate across a cell-free lane from one microglia lane to another if this distance of separation was less than about 60 [Formula: see text], indicating that propagation involved diffusion of a chemical transmitter. This transmitter was identified as ATP since all Ca(2 + ) wave propagation was blocked by the purinoceptor antagonist suramin, which blocks P2Y(2) and P2Y(12) at relatively low concentrations. Antibodies to P2Y(12) showed these at very high density compared with P2Y(2), indicating a role for P2Y(12) receptors. These observations were quantitatively accounted for by a model in which the main determinants are the diffusion of ATP released from a stimulated microglial cell and differences in the dissociation constant of the purinoceptors on the microglial cells.

Glutamate-stimulated ATP release from spinal cord astrocytes is potentiated by substance P.

ATP has recently emerged as a key molecule mediating pathological pain. The aim of this study was to examine whether spinal cord astrocytes could be a source of ATP in response to the nociceptive neurotransmitters glutamate and substance P. Glutamate stimulated ATP release from these astrocytes and this release was greatly potentiated by substance P, even though substance P alone did not elicit ATP release. Substance P also potentiated glutamate-induced inward currents, but did not cause such currents alone. When glutamate was applied alone it acted exclusively through alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionate receptors to stimulate Ca(2+) influx-dependent ATP release. However, when substance P was co-applied with glutamate, ATP release could be elicited by activation of NMDA and metabotropic glutamate receptors. Activation of neurokinin receptor subtypes, protein kinase C and phospholipases A(2), C and D were needed for substance P to bring about its effects. These results suggest that astrocytes may be a major source of ATP in the spinal cord on activation of nerve fibres that release substance P and glutamate.

Purinergic junctional transmission and propagation of calcium waves in spinal cord astrocyte networks.

Micro-photolithographic methods have been employed to form discrete patterns of spinal cord astrocytes that allow quantitative measurements of Ca(2+) wave propagation. Astrocytes were confined to lanes 20-100 microm wide and Ca(2+) waves propagated from a point of mechanical stimulation or of application of adenosine triphosphate; all Ca(2+) wave propagation was blocked by simultaneous application of purinergic P2Y(1) and P2Y(2) antagonists. Stimulation of an astrocyte at one end of a lane, followed by further stimulation of this astrocyte, gave rise to Ca(2+) transients in the same astrocytes; however, if the second stimulation was applied to an astrocyte at the other end of the lane, then this gave rise to a different but overlapping set of astrocytes generating a Ca(2+) signal. Both the amplitude and velocity of the Ca(2+) wave decreased over 270 microm from the point of initiation, and thereafter remained, on average, constant with random variations for at least a further 350 microm. Also, the percentage of astrocytes that gave a Ca(2+) transient decreased with distance along lanes. All the above observations were quantitatively predicted by our recent theoretical model of purinergic junctional transmission, as was the Ca(2+) wave propagation along and between parallel lanes of astrocytes different distances apart. These observations show that a model in which the main determinants are the diffusion of adenosine triphosphates regeneratively released from a stimulated astrocyte, together with differences in the properties and density of the purinergic P2Y receptors on astrocytes, is adequate to predict a wide range of Ca(2+) wave transmission and propagation phenomena.

Purine release from spinal cord microglia after elevation of calcium by glutamate.

The propagation of Ca2+ waves in a network of microglial cells, after its initiation by glutamate, is mediated by purinergic transmission. In this study, we investigated the mechanisms by which glutamate releases ATP from cultured spinal cord microglia. The 4-fold increase in ATP release from microglia in response to glutamate (0.5 mM) was blocked by alpha-aminohydroxy-5-methyl-isoxazole-4-proprionate (AMPA)/kainate receptor antagonist 6-cyano-7-nitroguinoxaline-2,3-dione and specific AMPA receptor antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466) but not by N-methyl-d-aspartic acid or metabotropic glutamate receptor antagonists. Glutamate acting on AMPA receptors evoked an ATP release that was blocked by antagonizing the rise in intracellular Ca2+ as a result of its release from internal stores as well as by antagonizing protein kinase C with chelerythrine. Glutamate-stimulated ATP release was significantly antagonized by the cystic fibrosis transmembrane conductance regulator (CFTR) blockers flufenamic acid and glibenclamide. A role for the CFTR was further confirmed using microglia from CFTR knockout mice, which released significantly less ATP than microglia from control wild-type mice in response to glutamate. Use of 6-methoxy-1-(3-sulfopropyl)quinolinium fluorescence assay revealed functional CFTR in microglia. These observations suggest that glutamate acted on microglial AMPA receptors to stimulate release of Ca2+ from intracellular stores as well as a Ca2+-dependent isoform of protein kinase C, which then acts to trigger release of ATP with the CFTR acting as a regulator of the ATP release process, perhaps through another channel or transporter.

Secretion of ATP from Schwann cells in response to uridine triphosphate.

The mechanisms by which uridine triphosphate (UTP) stimulates ATP release from Schwann cells cultured from the sciatic nerve were investigated using online bioluminescence techniques. UTP, a P2Y(2) and P2Y(4) receptor agonist, stimulated ATP release from Schwann cells in a dose-dependent manner with an ED(50) of 0.24 microm. UTP-stimulated ATP release occurs through P2Y(2) receptors as it was blocked by suramin which inhibits P2Y(2) but not P2Y(4) receptors. Furthermore, positive immunostaining of P2Y(2) receptors on Schwann cells was revealed and GTP, an equipotent agonist with UTP at rat P2Y(4) receptors, did not significantly stimulate ATP release. UTP-stimulated ATP release involved second messenger pathways as it was attenuated by the phospholipase C inhibitor U73122, the protein kinase C inhibitor chelerytherine chloride, the IP(3) formation inhibitor lithium chloride, the cell membrane-permeable Ca(2+) chelator BAPTA-AM and the endoplasmic reticulum Ca(2+)-dependent ATPase inhibitor thapsigargin. Evidence that ATP may be stored in vesicles that must be transported to the cell membrane for exocytosis was found as release was significantly reduced by the Golgi-complex inhibitor brefeldin A, microtubule disruption with nocodazole, F-actin disruption with cytochalasin D and the specific exocytosis inhibitor botulinum toxin A. ATP release from Schwann cells also involves anion transport as it was significantly reduced by cystic fibrosis transmembrane conductance regulator inhibitor glibencamide and anion transporter inhibitor furosemide. We suggest that UTP-stimulated ATP release is mediated by activation of P2Y(2) receptors that initiate an IP(3)-Ca(2+) cascade and protein kinase C which promote exocytosis of ATP from vesicles as well as anion transport of ATP across the cell membrane.

Mechanisms of secretion of ATP from cortical astrocytes triggered by uridine triphosphate.

The mechanisms involved in autocrine ATP release from cultured astrocytes isolated from the rat cortex were investigated using an online bioluminescence technique. Astrocytes released ATP in response to application of 10 microM uridine triphosphate, which was blocked by the non-specific purinergic receptor antagonist suramin. Intracellular pathways of the uridine triphosphate-stimulated ATP release were seen to involve inositol triphosphate and calcium with the assistance of the Golgi-complex and cytoskeleton as the release was inhibited by phospholipase C antagonist lithium, endoplasmic reticulum calcium-dependent ATPase inhibitor thapsigargin, F-actin interruptor cytochalasin D and Golgi-complex interruptor brefeldin A. The uridine triphosphate-stimulated ATP release was also potently blocked by exocytosis inhibitor botulinum toxin A and anion transporter blockers furosemide and glibenclamide. These results suggest that calcium-dependent exocytosis and transportation via anion transporters are the predominant secretion mechanisms for uridine triphosphate-stimulated ATP release from cortical astrocytes.

ATP secretion from nerve trunks and Schwann cells mediated by glutamate.

ATP release from rat sciatic nerves and from cultured Schwann cells isolated from the nerves was investigated using an online bioluminescence technique. ATP was released in relatively large amounts from rat sciatic nerve trunks during electrical stimulation. This release was blocked by the sodium channel inhibitor tetrodotoxin and the non-NMDA glutamate receptor blocker 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Schwann cells isolated from the nerve trunks did not release ATP when electrically stimulated but did in response to glutamate in a concentration-dependent manner. Glutamate-stimulated ATP release was inhibited by specific non-competitive AMPA receptor antagonist GYKI 52466 and competitive non-NMDA receptor antagonist CNQX. Glutamate-stimulated ATP release was decreased by inhibition of anion transporter inhibitors by furosemide, cystic fibrosis transmembrane conductance regulator by glibenclamide and exocytosis by botulinum toxin A, indicating that anion transporters and exocytosis provide the main secretion mechanisms for ATP release from the Schwann cells.