In mammals, receptors for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) are divided into three pharmacological classes, which are denoted GABAA, GABAB, and GABAC. GABAC receptors are defined by their insensitivity to the GABAA receptor antagonist bicuculline and the GABAB receptor agonist (-)-baclofen. GABAC receptors probably are a heterogeneous group of proteins. The most extensively studied mammalian GABAC receptors are those found in neurons of the outer retina. These receptors are GABA-gated Cl- channels comprised of p subunits, of which there are two subtypes. The physiological functions served by GABAC receptors are largely unknown; to determine the functions, it would be useful to have GABAC-selective ligands. In a previous study, we found that isoguvacine, a GABAA-selective agonist, and 3-aminopropyl-(methyl)phosphinic acid (3-APMPA), a GABAB-selective agonist, show affinity for retinal GABAC receptors. In particular, 3-APMPA is an antagonist with low micromolar potency (Kb approximately 1 microM). Here, we report the synthesis and pharmacological characterization of (1,2,5,6-tetrahydropyridine-4-yl)methylphosphinic acid (TPMPA), a hybrid of isoguvacine and 3-APMPA designed to retain affinity for GABAC receptors but not to interact with GABAA or GABAB receptors. Electrical assays show that TPMPA is a competitive antagonist of cloned human mu 1 GABAC receptors expressed in Xenopus laevis oocytes (Kb approximately 2 microM). TPMPA is > 100-fold weaker as an inhibitor of rat brain GABAA receptors expressed in oocytes (Kb approximately 320 microM) and has only weak agonist activity on GABAB receptors assayed in rat hippocampal slices (EC50 approximately 500 microM). TPMPA should be a useful pharmacological probe with which to investigate GABAC receptor function in the outer retina and in any other areas of the nervous system in which these types of receptor are present.

Cell membranes isolated from nervous tissue can be easily injected into Xenopus oocytes, thereby effectively "microtransplanting" functional neurotransmitter receptors. This technique therefore allows a direct functional characterization of the original membrane receptor/ion channel proteins and the associated molecules while still embedded in their natural lipid environment. Cell membranes will contain components from different types of cells, i.e. neurons and glial cells, expressing their own receptors, with possibly different properties. To study the receptor properties of a single cell type, we injected oocytes with membranes isolated only from glia (gliosomes) of adult mouse neocortex and we focused our work on GABA(A) receptors incorporated in the oocyte cell membrane. We found that GABA(A)-activated currents allowed a good biophysical and pharmacological characterization of glial GABA(A) receptors. Therefore, the microtransplantation of gliosomes into oocytes can represent a good model to study the electrical and pharmacological properties of adult glial cells under different physiological and pathological conditions. Moreover, since gliosomes can be isolated from frozen tissues, this approach can be extended to post-mortem human tissues.

The chemokine CX3CL1 and its receptor CX3CR1 are constitutively expressed in the nervous system. In this study, we used in vivo murine models of permanent middle cerebral artery occlusion (pMCAO) to investigate the protective potential of CX3CL1. We report that exogenous CX3CL1 reduced ischemia-induced cerebral infarct size, neurological deficits, and caspase-3 activation. CX3CL1-induced neuroprotective effects were long lasting, being observed up to 50 d after pMCAO in rats. The neuroprotective action of CX3CL1 in different models of brain injuries is mediated by its inhibitory activity on microglia and, in vitro, requires the activation of adenosine receptor 1 (A1R). We show that, in the presence of the A1R antagonist 1,3-dipropyl-8-cyclopentylxanthine and in A1R−/− mice, the neuroprotective effect of CX3CL1 on pMCAO was abolished, indicating the critical importance of the adenosine system in CX3CL1 protection also in vivo. In apparent contrast with the above reported data but in agreement with previous findings, cx3cl1−/− and cx3cr1GFP/GFP mice, respectively, deficient in CX3CL1 or CX3CR1, had less severe brain injury on pMCAO, and the administration of exogenous CX3CL1 increased brain damage in cx3cl1−/− ischemic mice. We also report that CX3CL1 induced a different phagocytic activity in wild type and cx3cl1−/− microglia in vitro during cotreatment with the medium conditioned by neurons damaged by oxygen-glucose deprivation. Together, these data suggest that acute administration of CX3CL1 reduces ischemic damage via an adenosine-dependent mechanism and that the absence of constitutive CX3CL1–CX3CR1 signaling changes the outcome of microglia-mediated effects during CX3CL1 administration to ischemic brain.

Fractalkine/CX3CL1 is a neuron-associated chemokine, which modulates microglia-induced neurotoxicity activating the specific and unique receptor CX3CR1. CX3CL1/CX3CR1 interaction modulates the release of cytokines from microglia, reducing the level of tumor necrosis factor-α, interleukin-1-β, and nitric oxide and induces the production of neurotrophic substances, both in vivo and in vitro. We have recently shown that blocking adenosine A1 receptors (A1R) with the specific antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) abolishes CX3CL1-mediated rescue of neuronal excitotoxic death and that CX3CL1 induces the release of adenosine from microglia. In this study, we show that the presence of extracellular adenosine is mandatory for the neurotrophic effect of CX3CL1 as reducing adenosine levels in hippocampal cultures, by adenosine deaminase treatment, strongly impairs CX3CL1-mediated neuroprotection. Furthermore, we confirm the predominant role of microglia in mediating the neuronal effects of CX3CL1, because the selective depletion of microglia from hippocampal cultures treated with clodronate-filled liposomes causes the complete loss of effect of CX3CL1. We also show that hippocampal neurons obtained from A1R−/− mice are not protected by CX3CL1 whereas A2AR−/− neurons are. The requirement of functional A1R for neuroprotection is not unique for CX3CL1 as A1R−/− hippocampal neurons are not rescued from Glu-induced cell death by other neurotrophins such as brain-derived neurotrophic factor and erythropoietin, which are fully active on wt neurons.

Verapamil, a Ca2+ channel blocker widely used in clinical practice, also affects the properties of frog and mouse muscle acetylcholine receptor (AChR). Here, we examine the mechanism of action of verapamil on human wild-type and slowchannel mutant muscle AChRs harboring in any subunit a valine-to-alanine mutation of 13' residue of the pore-lining M2 transmembrane segment. Verapamil, after a pre-treatment of 0.5-10 s, accelerated the decay of whole-cell or macroscopic outside-out currents within milliseconds of ACh application even at clinically attainable doses. Recordings of unitary events in the cell-attached and outside-out configurations showed that verapamil does not alter single-channel conductance, but reduces channel open probability, by prolonging the dwell time into the closed state for wild-type and all mutant AChR. The duration of channel openings decreased only for the epsilon V265A-AChR, by shortening the longest exponential component of the open-time distribution. These results provide a rationale for the therapeutic use of verapamil in the slow-channel syndrome and emphasize the major role played by epsilon subunit in controlling the functional properties of human muscle AChR, as revealed by the peculiar alterations imparted by mutations in this subunit.

Here we tested two working hypotheses on spatially selective cortical activation (“neural efficiency”) in experts: (i) compared to non-athletes, elite karate athletes are characterized by a reduced cortical activation during the judgment of karate actions; (ii) compared to non-athletes and elite karate athletes, amateur karate athletes are characterized by an intermediate cortical activation during the judgment of karate actions. Electroencephalographic (EEG) data were recorded in 16 elite karate athletes, 15 amateur athletes and 17 non-athletes. They observed a series of 120 karate videos. At the end of each video, the subjects had to judge the technical/athletic level of the exercise by a scale from 0 to 10. The mismatch between their judgment and that of the coach indexed the degree of action judgment. The EEG cortical sources were estimated by sLORETA. With reference to a pre-stimulus period, the power decrease of alpha (8–12Hz) rhythms during the video indexed the cortical activation (event-related desynchronization, ERD). Regarding the hypothesis of reduced activity in elite karate athletes, low- and high-frequency alpha ERD was less pronounced in dorsal and “mirror” pathways in the elite karate athletes than in the non-athletes. Regarding the hypothesis of intermediate cortical activity in amateur karate athletes, low- and high-frequency alpha ERD was less pronounced in dorsal pathways across the non-athletes, the amateur karate athletes, and the elite karate athletes. In conclusion, athletes’ judgment of observed sporting actions is related to less pronounced alpha ERD, as a possible index of “neural efficiency” in experts engaged in social cognition.

Objective: Previous evidence in epileptic subjects has shown that theta (about 4-7 Hz) and gamma rhythms (about 40-45 Hz) of hippocampus, amygdala, and neocortex were temporally synchronized during the listening of repeated words successfully remembered (Babiloni et al., 2009). Here we re-analyzed those electroencephalographic (EEG) data to test whether a parallel increase in amplitude of late positive event-related potentials takes place.Methods: Intracerebral electroencephalographic (EEG) activity had been recorded in five subjects with drug-resistant temporal lobe epilepsy, undergoing pre-surgical evaluation. During the recording of the intracerebral EEG activity, the subjects performed a computerized version of the Rey auditory verbal learning test (RAVLT). They heard the same list of 15 common words for five times. Each time, immediately after the listening of the list, the subjects were required to repeat as many words as they could recall.Results: We found that late positive event-related potentials (ERPs) peaking at about 350 ms post-stimulus in amygdala, hippocampus, and occipital-temporal cortex had a higher amplitude during the listening of the repeated words that were subsequently recalled than for those that were not recalled.Conclusions: Late positive ERPs reflect a functional mechanism implemented in a human brain network spanning amygdala, hippocampus, and occipital-temporal cortex which is at the basis of the memorization processes of verbal materials.Significance: This ERP component is a promising neuromarker of successful memorization of repeated words in humans. (C) 2010 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

The present electroencephalographic (EEG) study tested the working hypothesis that the amplitude of resting state cortical EEG rhythms (especially alpha, 8-12 Hz) was higher in elite athletes compared with amateur athletes and non-athletes, as a reflection of the efficiency of underlying back-ground neural synchronization mechanisms. Eyes closed resting state EEG data were recorded in 16 elite karate athletes, 20 amateur karate athletes, and 25 non-athletes. The EEG rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). EEG cortical sources were estimated by low-resolution brain electromagnetic tomography (LORETA). Statistical results showed that the amplitude of parietal and occipital alpha 1 sources was significantly higher in the elite karate athletes than in the non-athletes and karate amateur athletes. Similar results were observed in parietal and occipital delta sources as well as in occipital theta sources. Finally, a control confirmatory experiment showed that the amplitude of parietal and occipital delta and alpha 1 sources was stronger in 8 elite rhythmic gymnasts compared with 14 non-athletes. These results supported the hypothesis that cortical neural synchronization at the basis of eyes-closed resting state EEG rhythms is enhanced in elite athletes than in control subjects.

We applied the group-I metabotropic glutamate (mGlu) receptor agonist, 3,5-dihydroxyphenylglycine (DHPG), to neonatal or adult rat hippocampal slices at concentrations (10 μm) that induced a short-term depression (STD) of excitatory synaptic transmission at the Schaffer collateral/CA1 synapses. DHPG-induced STD was entirely mediated by the activation of mGlu5 receptors because it was abrogated by the mGlu5 receptor antagonist, MPEP [2-methyl-6-(phenylethynyl)pyridine], but not by the mGlu1 receptor antagonist, CPCCOEt [7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester]. Knowing that ephrin-Bs functionally interact with group-I mGlu receptors (Calò et al., 2005), we examined whether pharmacological activation of ephrin-Bs could affect DHPG-induced STD. We activated ephrin-Bs using their cognate receptor, EphB1, under the form of a preclustered EphB1/Fc chimera. Addition of clustered EphB1/Fc alone to the slices induced a small but nondecremental depression of excitatory synaptic transmission, which differed from the depression induced by 10 μmDHPG. Surprisingly, EphB1/Fc-induced synaptic depression was abolished by MPEP (but not by CPCCOEt) suggesting that it required the endogenous activation of mGlu5 receptors. In addition, coapplication of DHPG and EphB1/Fc, resulted in a large and nondecremental long-term depression. The effect of clustered EphB1/Fc was specific because it was not mimicked by unclustered EphB1/Fc or clustered EphA1/Fc. These findings raise the intriguing possibility that changes in synaptic efficacy mediated by mGlu5 receptors are under the control of the ephrin/Eph receptor system, and that the neuronal actions of ephrins can be targeted by drugs that attenuate mGlu5 receptor signaling.

Objective: The "neural efficiency" hypothesis posits that neural activity is reduced in experts. Here we tested the hypothesis that compared with non-athletes, elite athletes are characterized by a reduced cortical activation during simple voluntary movement and that this is reflected by the modulation of dominant alpha rhythms (8-12 Hz).Methods: EEG data (56 channels; EB-Neuro) were continuously recorded in the following right-handed subjects: 10 elite karate athletes and 12 non-athletes. During the EEG recordings, they performed brisk voluntary wrist extensions of the right or left hand (right movement and left movement). The EEG cortical sources were estimated by standardized low-resolution brain electromagnetic tomography (sLORETA) freeware. With reference to a baseline period, the power decrease of alpha rhythms during the motor preparation and execution indexed the cortical activation (event-related desynchronization, ERD).Results: During both preparation and execution of the right movements, the low-(about 8-10 Hz) and high-frequency alpha ERD (about 10-12 Hz) was lower in amplitude in primary motor area, in lateral and medial premotor areas in the elite karate athletes than in the non-athletes. For the left movement, only the high-frequency alpha ERD during the motor execution was lower in the elite karate athletes than in the non-athletes.Conclusions: These results confirmed that compared with non-athletes, elite athletes are characterized by a reduced cortical activation during simple voluntary movement. Significance: Cortical alpha rhythms are implicated in the "neural efficiency" of athletes' motor systems. (C) 2009 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

We previously found that the endogenous anticonvulsant adenosine, acting through A 2A and A 3 adenosine receptors (ARs), alters the stability of currents (I GABA ) generated by GABA A receptors expressed in the epileptic human mesial temporal lobe (MTLE). Here we examined whether ARs alter the stability (desensitization) of I GABA expressed in focal cortical dysplasia (FCD) and in periglioma epileptic tissues. The experiments were performed with tissues from 23 patients, using voltage-clamp recordings in Xenopus oocytes microinjected with membranes isolated from human MTLE and FCD tissues or using patch-clamp recordings of pyramidal neurons in epileptic tissue slices. On repetitive activation, the epileptic GABA A receptors revealed instability, manifested by a large I GABA rundown, which in most of the oocytes (≈70%) was obviously impaired by the new A 2A antagonists ANR82, ANR94, and ANR152. In most MTLE tissue-microtransplanted oocytes, a new A 3 receptor antagonist (ANR235) significantly improved I GABA stability. Moreover, patch-clamped pyramidal neurons from human neocortical slices of periglioma epileptic tissues exhibited altered I GABA rundown on ANR94 treatment. Our findings indicate that antagonizing A 2A and A 3 receptors increases the I GABA stability in different epileptic tissues and suggest that adenosine derivatives may offer therapeutic opportunities in various forms of human epilepsy.

“Neural efficiency” hypothesis posits that neural activity is reduced in experts. Here we tested the hypothesis that compared with non-athletes, elite athletes are characterized by a reduction of cortical activation during an engaging upright standing. EEG (56 channels; Be-plus Eb-Neuro©) and stabilogram (RGM©) data were simultaneously recorded in 10 elite karate, 10 elite fencing athletes, and 12 non-athletes during a simple bipodalic (standard Romberg) and a more engaging monopodalic upright standing. Balance was indexed by body “sway area”. The EEG data were spatially enhanced by surface Laplacian estimation. Cortical activity was indexed by task-related power decrease (TRPD) of EEG alpha power (8–12 Hz) during monopodalic referenced to bipodalic condition. The body “sway area” was larger during the monopodalic than bipodalic upright standing in all groups. Low-frequency alpha TRPD (about 8–10 Hz) was lower in amplitude in the karate and fencing athletes than in the non-athletes at left central, right central, middle parietal, and right parietal areas (p < 0.01). Similarly, the amplitude of high-frequency alpha TRPD (10–12 Hz) was lower in the karate and fencing athletes than in the non-athletes at right frontal, left central, right central, and middle parietal areas (p < 0.03). These results suggest that during monopodalic referenced to less engaging bipodalic condition, the power decrease (i.e. the desynchronization) of cortical activity at alpha rhythms is largely reduced in elite athletes than in non-athletes, in line with the “neural efficiency” hypothesis. The present study extends our understanding of the physiological mechanisms at the basis of the “neural efficiency” for engaging upright standing in elite athletes.

Objective: Non-steroidal anti-inflammatory drugs such as ibuprofen have a protective role on risk of Alzheimer's disease (AD). Here we evaluated the hypothesis that long-term ibuprofen treatment affects cortical Sources of testing electroencephalographic (EEG) rhythms in mild AD patients.Methods: Twenty-three AD patients (13 treated AD IBUPROFEN: 10 untreated AD PLACEBO) were enrolled. Resting EEG data were recorded before and 1 year after the ibuprofen/placebo treatment. EEG rhythms were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (1320 Hz), and beta 2 (20-30 Hz). LORETA was used for EEG source analysis.Results: In the AD PLACEBO group, amplitude of delta Sources was globally greater at follow-up than baseline. Instead, amplitude of delta Sources remained stable or decreased in the majority of the AD IBUPROFEN patients. Clinical (CDR) but not global cognitive Status (MMSE) reflected EEG results.Conclusions: These results suggest that in mild AD patients, a long-term ibuprofen treatment slightly slows down the progressive increment of delta rhythms as a sign of contrast against the neurodegenerative processes.Significance: They motivate future investigations with larger population and extended neuropsychological testing, to study the relationships among ibuprofen treatment, delta cortical Sources, and higher order functions. (C) 2009 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.

This study tested the two following hypotheses: (i) compared with non‐athletes, elite athletes are characterized by a reduced cortical activation during the preparation of precise visuo‐motor performance; (ii) in elite athletes, an optimal visuo‐motor performance is related to a low cortical activation. To this aim, electroencephalographic (EEG; 56 channels; Be Plus EB‐Neuro) data were recorded in 18 right‐handed elite air pistol shooters and 10 right‐handed non‐athletes. All subjects performed 120 shots. The EEG data were spatially enhanced by surface Laplacian estimation. With reference to a baseline period, power decrease/increase of alpha rhythms during the preshot period indexed the cortical activation/deactivation (event‐related desynchronization/synchronization, ERD/ERS). Regarding the hypothesis (i), low‐ (about 8–10 Hz) and high‐frequency (about 10–12 Hz) alpha ERD was lower in amplitude in the elite athletes than in the non‐athletes over the whole scalp. Regarding the hypothesis (ii), the elite athletes showed high‐frequency alpha ERS (about 10–12 Hz) larger in amplitude for high score shots (50%) than for low score shots; this was true in right parietal and left central areas. A control analysis confirmed these results with another indicator of cortical activation (beta ERD, about 20 Hz). The control analysis also showed that the amplitude reduction of alpha ERD for the high compared with low score shots was not observed in the non‐athletes. The present findings globally suggest that in elite athletes (experts), visuo‐motor performance is related to a global decrease of cortical activity, as a possible index of spatially selective cortical processes (“neural efficiency”). Hum Brain Mapp, 2009. © 2009 Wiley‐Liss, Inc.

Atrophy of hippocampus and alteration of resting eyes-closed electroencephalographic (EEG) rhythms represent important features of mild cognitive impairment (MCI) and Alzheimer's disease (AD). Here we evaluated linear and non-linear aspects of the relationship between these features in the continuum along MCI and AD conditions, as a reflection of neurodegenerative processes. Eyes-closed resting EEG data were recorded in 60 healthy elderly (Nold), 88 MCI, and 35 Alzheimer's disease (AD) patients. Hippocampal volume was measured in magnetic resonance imaging of the MCI and AD subjects. Based on the normalized hippocampal volume, selected MCI subjects could be divided into two demographically paired sub-groups: those with larger hippocampal volume (MCI + h; N = 40; mini mental state evaluation - MMSE - score = 27.5 +/- 0.26 SE) and those with smaller hippocampal volume (MCI - h; N = 40; h; MMSE = 26.5 +/- 0.34 SE); the normalized hippocampal volume was statistically greater in the MCI + h than in the MCI - h and AD subjects (p<0.0001). EEG rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). Cortical EEG generators were estimated by LORETA software. Results showed that the power of occipital, parietal, and temporal alpha 1 sources was maximum in MCI + h, intermediate in MCI - h, and low in AD patients. Furthermore, the power of these sources was linearly and non-linearly correlated with the normalized hippocampal volume. These 3 EEG sources were given as input for evaluating correlations (linear, exponential, logarithmic and power) with hippocampal volume. When subjects were considered as a unique group, there was a significant linear correlation of hippocampal volume with the magnitude of alpha 1 sources in the parietal, occipital and temporal areas. In general, the EEG sources showing significant linear correlation with hippocampal volume also supported a non-linear correlation with hippocampal volume strongly for the logarithmic one. The present results suggest that progressive atrophy of hippocampus correlates with decreased cortical alpha power, as estimated by using LORETA source modeling, in the continuum along MCI and AD conditions. (C) 2008 Elsevier Inc. All rights reserved.

OBJECTIVE:Are obese subjects characterized by a reduction of attentional cortical responses to the enlargement of food or body images? METHODS:Electroencephalographic data were recorded in 19 obese and 15 normal-weight adults during an "oddball" paradigm. The subjects were given frequent (70%) and rare (30%) stimuli depicting faces (FACE), food (FOOD), and landscapes (CONTROL), and clicked the mouse after the rare stimuli. These stimuli depicted the same frequent stimuli graphically dilated by 25% along the horizontal axis. Bioelectrical impedance indexed subjects' body fat percentage. Cortical attentional responses were probed by the difference between positive event-related potentials peaking around 400-500ms post-stimulus for the rare minus frequent stimuli (P300). Low resolution electromagnetic source tomography (LORETA) estimated P300 sources. RESULTS:In the FOOD condition, the amplitude of medial prefrontal P300 sources (Brodmann area 9) was lower in the obese than normal-weight subjects, and there was a negative correlation between the body fat percentage and the amplitude of these sources in all subjects as a single group. CONCLUSIONS:These results disclose that prefrontal attentional processes to food size are abnormal in obese subjects. SIGNIFICANCE:The present study motivates future research evaluating the effects of cognitive rehabilitation in obese subjects.