Biomedical subjects

G Biella

Publications and source records attributed to G Biella.

At least 19 recordsLinked to original sources

PubMed2005

Caveolin-1 expression and membrane cholesterol content modulate N-type calcium channel activity in NG108-15 cells.

Caveolins are the main structural proteins of glycolipid/cholesterol-rich plasmalemmal invaginations, termed caveolae. In addition, caveolin-1 isoform takes part in membrane remodelling as it binds and transports newly synthesized cholesterol from endoplasmic reticulum to the plasma membrane. Caveolin-1 is expressed in many cell types, including hippocampal neurons, where an abundant SNAP25-caveolin-1 complex is detected after induction of persistent synaptic potentiation. To ascertain whether caveolin-1 influences neuronal voltage-gated Ca2+ channel basal activity, we stably expressed caveolin-1 into transfected neuroblastoma x glioma NG108-15 hybrid cells [cav1(+) clone] that lack endogenous caveolins but express N-type Ca2+ channels upon cAMP-induced neuronal differentiation. Whole-cell patch-clamp recordings of cav1(+) cells demonstrated that N-type current density was reduced in size by approximately 70% without any significant change in the time course of activation and inactivation and voltage dependence. Moreover, the cav1(+) clone exhibited a significantly increased proportion of membrane cholesterol compared to wild-type NG108-15 cells. To gain insight into the mechanism underlying caveolin-1 lowering of N-current density, and more precisely to test whether this was indirectly caused by caveolin-1-induced enhancement of membrane cholesterol, we compared single N-type channel activities in cav1(+) clone and wild-type NG108-15 cells enriched with cholesterol after exposure to a methyl-beta-cyclodextrin-cholesterol complex. A lower Ca2+ channel activity was recorded from cell-attached patches of both cell types, thus supporting the view that the increased proportion of membrane cholesterol is ultimately responsible for the effect. This is due to a reduction in the probability of channel opening caused by a significant decrease of channel mean open time and by an increase of the frequency of null sweeps.

Animals↗

PubMed2004

Evaluation of three quinoline-carboxamide derivatives as potential radioligands for the in vivo pet imaging of neurodegeneration.

The peripheral-type benzodiazepine receptors (PBRs) are only minimally expressed in normal brain parenchyma, where they are primarily localized in glial cells. Their basal expression rises in different neurodegenerative disorders, due to the presence of infiltrating inflammatory cells and activated microglia. [11C]PK11195, a selective PBR antagonist, has been used for the in vivo PET monitoring of neurodegeneration in clinical observations. We recently developed and labeled with carbon-11 three new carboxamide derivatives: [11C]VC193M, [11C]VC195 and [11C]VC198M. Aim of this study was to evaluate these ligands for the in vivo measuring of PBRs expression in neurodegenerations and compare their kinetic behavior with that of the reference tracer [11C]PK11195. Radioligands were evaluated in a preclinical model of Huntington's disease consisting in the monolateral striatal injection of quinolinic acid (QA). Activated microglia and astrocytic gliosis was present only within the affected striatum. A concomitant increase in radioactivity accumulation was observed for all the tracers examined (P<0.01). Among the new compounds, [11C]VC195 showed higher levels of lesioned/unlesioned striatum ratios (3.28+/-0.44), in comparison with [11C]VC193M and [11C]VC198M (2.69+/-0.53 and 1.52+/-0.36, respectively), but slightly inferior to that observed for [11C]PK11195 (3.76+/-1.41).In conclusion, the results of the study indicate that [11C]VC195 is a promising candidate for in vivo PET monitoring of neurodegenerative processes but its in vivo behavior overlap that of [11C]PK11195.

Amides↗

PubMed2002

Nefopam, an analogue of orphenadrine, protects against both NMDA receptor-dependent and independent veratridine-induced neurotoxicity.

Nefopam hyghochloride is a potent analgesic compound commercialized in most Western Europe for 20 years, which possesses a profile distinct from that of opioids or anti-inflammatory drugs. Previous evidence suggested a central action of nefopam but the detailed mechanisms remain unclear. While, nefopam structure resembles that of orphenadrine, an uncompetitive NMDA receptor antagonist, here we report that differently from orphenadrine, nefopam (100 microM) failed to protect cultured cerebellar neurons from excitotoxicity following direct exposure of neurons to glutamate. Moreover, nefopam failed to displace MK-801 binding to hippocampal membranes. Nefopam effectively prevented NMDA receptor-mediated early appearance (30 min) of toxicity signs induced by the voltage sensitive sodium channel (VSSC) activator veratridine. The later phase (24 h) of neurotoxicity by veratridine occurring independently from NMDA receptor activation, was also prevented by nefopam. Nefopam effect was not mimicked by the GABA receptor agonist muscimol.

Analgesics, Non-Narcotic↗

PubMed2001

Contralateral treatment with lidocaine reduces spinal neuronal activity in mononeuropathic rats.

In anaesthetised and paralysed rats with chronic constriction of the sciatic nerve, the effects of subcutaneous contralateral lidocaine (100 microl) on the activity of lumbar (L(4)-L(5)) wide dynamic range neurons ipsilateral to the constriction have been investigated. The results show reduction of the spontaneous hyperactivity for 60 min; suppression or reduction of the responses to contralateral noxious stimulation for 60 min; lack of effect on the responses to ipsilateral noxious stimulation, except for the afterdischarge duration, reduced for 60 min. The finding that the altered neuronal activity following peripheral nerve injury associated to behavioural signs of neuropathic pain, can be reduced by contralateral treatment, may provide further suggestions to neuropathic pain mechanisms and management.

Action Potentials↗

PubMed2001

Effect of different concentrations of iontophoretic nociceptin on distinct classes of nociceptive neurons in rat spinal cord.

Iontophoretically applied nociceptin (NC) was tested at different concentrations on the activity of spinal nociceptive specific (NS) and wide dynamic range (WDR) neurons. Low NC dosages inhibited the noxious response of NS neurons, higher dosages inhibited the noxious responses of the WDR neurons but had little effect on the non-noxious response. Naloxone did not antagonize the NC effect. Thus, appropriate dosages of NC may be selective, both for neuronal classes and for sensory modalities.

Animals↗

PubMed2001

Acupuncture produces central activations in pain regions.

Acupuncture is largely used for pain control in several pathological conditions. Its effects on the central nervous system are not well defined. We investigated the effect of the application of acupuncture to 13 normal subjects (males, 21-32 years). H2(15)O bolus PET scans were read before the application of the needles (Rest, R) and after 25 min of needle insertion. Data were acquired by scanning in 3-D mode. The acupuncture application, true acupuncture (TA), was alternated to a placebo needle application (PA) in two different sequences (seven and six subjects, respectively), either R,PA,R, TA or R,TA,R,PA, a period of 15 min being left after every first TA or PA to allow for the recovery of basal conditions. Here we show that classic acupuncture activates the left Anterior Cingulus, the Insulae bilaterally, the Cerebellum bilaterally, the left Superior Frontal Gyrus, and the right Medial and Inferior Frontal Gyri. Most of the activated areas are shared with areas activated in acute and chronic pain states as described in the literature. Thus acupuncture appears to act by activating areas also involved in pain. This indicates that acupuncture could relief pain by unbalancing the equilibrium of distributed pain-related central networks.

Acupuncture Points↗

PubMed2001

Novel effect of nefopam preventing cGMP increase, oxygen radical formation and neuronal death induced by veratridine.

Nefopam hydrochloride is a potent analgesic compound that possesses a profile distinct from that of opiods or anti-inflammatory drugs. Previous evidence suggested a central action of nefopam but the detailed mechanisms remain unclear. Here we have used cultured cerebellar neurons to test the hypothesis that nefopam may modulate voltage sensitive sodium channel (VSSC) activity. Nefopam (100 microM) effectively prevented NMDA receptor-mediated early appearance (30 min) of toxicity signs induced by the VSSC activator veratridine. Delayed neurotoxicity by veratridine occurring independently from NMDA receptor activation, was also prevented by nefopam. In contrast, excitotoxicity following direct exposure of neurons to glutamate was not affected. Neuroprotection by nefopam was dose-dependent. 50% protection was obtained at 57 microM while full neuroprotection was achieved at 75 microM nefopam. Veratridine-induced sodium influx was completely abolished in nefopam-treated neurons. Intracellular cGMP and oxygen radical formation following VSSC stimulation by veratridine were also effectively prevented by nefopam. Our data are consistent with an inhibitory action of nefopam on VSSC and suggest that nefopam may modulate the release of endogenous glutamate following activation of these channels. This novel action of nefopam may be of great interest for the treatment of neurodegenerative disorders involving excessive glutamate release and neurotransmission.

Analgesics, Non-Narcotic↗

PubMed2001

Discharge threshold is enhanced for several seconds after a single interictal spike in a model of focal epileptogenesis.

Interictal spikes (ISs) are typically observed between seizures in focal epilepsies. Whether ISs are causally involved or represent protective elements in the transition toward an ictal discharge is an open question. Previous studies suggested that inhibition or disfacilitation occurs during the period elapsing between two ISs induced by local application of either bicuculline or penicillin in the piriform cortex of the in vitro isolated guinea pig brain preparation. We further investigated this issue by studying responses to afferent stimulation during the interspike period (6.3 +/- 2.5 s; mean +/- SD). Properly set stimulation intensity of the lateral olfactory tract resets ISs exclusively (and not before) 4-10 s (5.6 +/- 2.0 s; mean +/- SD) after a preceding spontaneous spike. This finding demonstrates the existence of a period of enhanced threshold to stimulus-evoked activation that coincides with the interspike interval in the absence of stimulation. Current source density analysis of depth laminar profiles demonstrated that both stimulus-evoked and spontaneous ISs were generated by the activation of an identical cortical circuit. Our study suggests that interictal spiking could play a protective role or at least provide an effective restraint against the onset of a focal ictal discharge.

Action Potentials↗

PubMed2001

Network activity evoked by neocortical stimulation in area 36 of the guinea pig perirhinal cortex.

The perirhinal cortex is a key structure involved in memory consolidation and retrieval. In spite of the extensive anatomical studies that describe the intrinsic and extrinsic associative connections of the perirhinal cortex, the activity generated within such a network has been poorly investigated. We describe here the pattern of synaptic interactions that subtend the responses evoked in area 36 of the perirhinal cortex by neocortical and local stimulation. The experiments were carried out in the in vitro isolated guinea pig brain. The synaptic perirhinal circuit was reconstructed by integrating results obtained during intracellular recordings from layer II-III neurons with simultaneous current source density analysis of laminar profiles performed with 16-channel silicon probes. Both neocortical and local stimulation of area 36 determined a brief monosynaptic excitatory potential in layer II-III neurons, followed by a biphasic synaptic inhibitory potential possibly mediated by a feed-forward inhibitory circuit at sites close to the stimulation electrode and a late excitatory postsynaptic potential (EPSP) that propagated at distance within area 36 along the rhinal sulcus. During a paired-pulse stimulation test, the inhibitory postsynaptic potential (IPSP) and the late EPSP were abolished in the second conditioned response, suggesting that they are generated by poli-synaptic circuits. Current source density analysis of the field responses demonstrated that 1) the monosynaptic activity was generated in layers II-III and 2) the sink associated to the disynaptic responses was localized within the superficial layer of area 36. We conclude that the neocortical input induces a brief monosynaptic excitation in area 36 of the perirhinal cortex, that is curtailed by a prominent inhibition and generates a recurrent excitatory associative response that travels at distance within area 36 itself. The results suggest that the perirhinal cortex network has the potentials to integrate multimodal incoming neocortical information on its way to the hippocampus.

Animals↗

PubMed2000

Evidence for spatial modules mediated by temporal synchronization of carbachol-induced gamma rhythm in medial entorhinal cortex.

Fast (gamma) oscillations in the cortex underlie the rapid temporal coordination of large-scale neuronal assemblies in the processing of sensory stimuli. Cortical gamma rhythm is modulated in vivo by cholinergic innervation from the basal forebrain and can be generated in vitro after exogenous cholinergic stimulation. Using the isolated guinea pig brain, an in vitro preparation that allows for the study of an intact cerebrum, we studied the spatial features of gamma activity evoked by the cholinomimetic carbachol (CCh) in the medial entorhinal cortex (mEC). gamma activity induced by either arterial perfusion or intraparenchymal application of CCh showed a phase reversal across mEC layer II and was reduced or abolished in a spatially localized region by focal infusions of atropine, bicuculline, and CNQX. In addition, a spatially restricted zone of gamma activity could be induced by passive diffusion of CCh from a recording pipette. Finally, gamma oscillations recorded at multiple sites across the surface of the mEC using array electrodes during arterial perfusion of CCh demonstrated a decline in synchronization (coherence) as the interelectrode distance increased. This effect was independent of the signal amplitude and was specific for gamma as opposed to theta-like activity induced by CCh in the same experiments. These results suggest that CCh-induced gamma oscillations in the mEC are mediated through direct muscarinic excitation of a highly localized reciprocal inhibitory-excitatory network located in superficial layers. We propose that functional cortical modules of highly synchronous gamma oscillations may organize incoming (cortical) and outgoing (hippocampal) information in the mEC.

Animals↗

PubMed2000

Differential effects of MK-801, a N-methyl-D-aspartate non-competitive antagonist, on the dorsal horn neuron hyperactivity and hyperexcitability in neuropathic rats.

N-methyl-D-aspartate (NMDA) involvement in altered spinal neuron activity following peripheral nerve injury has been investigated in rats with chronic constriction of the sciatic nerve. Extracellular single neuron recordings were performed, in anesthetized, paralyzed rats, from the sciatic spinal cord segments (lumbar, L5-L6) ipsilateral to the constriction, and the effect of iontophoresized MK-801, an NMDA receptor non-competitive antagonist, was tested on baseline hyperactivity and hyperresponsiveness to noxious stimulation. The results show that baseline activity was unaffected whereas the noxious evoked responses were significantly modified, there being amplitude reduction and after-discharges suppression. The different role of NMDA in the abnormal pain states related to the abnormal neuronal activities is discussed.

Animals↗

PubMed2000

Contribution of central sensitization to the pain-related abnormal activity in neuropathic rats.

The possibility of different contributions from peripheral and central sensitization to distinct neuropathic pain syndromes has been studied in rats with chronic constriction of the sciatic nerve (CCI), showing positive behavioral signs of neuropathic pain. In anesthetized, paralyzed rats extracellular recordings were performed in the spinal sciatic afferent territory (L5-L6), ipsilateral to the injured nerve, from wide dynamic range (WDR) neurons. The spontaneous activity and the responses to noxious stimuli applied to the proper area, i.e., the skin innervated by the constricted sciatic nerve, and to "inappropriate" areas, like the tail and the area of skin supplied by the contralateral sciatic and saphenous nerves, were analyzed before and after input from the constricted nerve was reversibly blocked at the ganglionic level by local anesthetic. The neurons discharged spontaneously with high frequencies, and responded to the stimulation of proper and "inappropriate" areas with high frequency discharge and prolonged afterdischarges During the ganglionic block, confirmed by the lack of responses to proper area stimulation, theWDR neuron background activity was significantly reduced; the responses to all "inappropriate" afferences were present, the frequency discharges being comparable to the preblock ones while the afterdischarges were significantly shorter. Since the efficacy of "inappropriate" inputs is related to neuronal sensitization, the persistence of these responses indicates that central neurons remain sensitized during peripheral block. In view of the relationship between the examined spontaneous and stimulated activities and neuropathic pain symptoms, the data suggest that central sensitization contributes with different drive strength to such symptoms, playing a crucial role in extraterritorial pain.

Afferent Pathways↗

PubMed2000

Olfactory inputs activate the medial entorhinal cortex via the hippocampus.

The lateral and medial regions of the entorhinal cortex differ substantially in terms of connectivity and pattern of activation. With regard to olfactory input, a detailed and extensive physiological map of the olfactory projection to the entorhinal cortex is missing, even if anatomic studies suggest that the olfactory afferents are confined to the lateral and rostral entorhinal region. We studied the contribution of the medial and lateral entorhinal areas to olfactory processing by analyzing the responses induced by lateral olfactory tract stimulation in different entorhinal subfields of the in vitro isolated guinea pig brain. The pattern of synaptic activation of the medial and lateral entorhinal regions was reconstructed either by performing simultaneous multisite recordings or by applying current source density analysis on field potential laminar profiles obtained with 16-channel silicon probes. Current source density analysis demonstrated the existence of a direct monosynaptic olfactory input into the superficial 300 microm of the most rostral part of the lateral entorhinal cortex exclusively, whereas disynaptic sinks mediated by associative fibers arising from the piriform cortex were observed at 100-350 microm depth in the entire lateral aspect of the cortex. No local field responses were recorded in the medial entorhinal region unless a large population spike was generated in the hippocampus (dentate gyrus and CA1 region) by a stimulus 3-5x the intensity necessary to obtain a maximal monosynaptic response in the piriform cortex. In these conditions, a late sink was recorded at a depth of 600-1000 microm in the medial entorhinal area (layers III-V) 10.6 +/- 0.9 (SD) msec after a population spike was simultaneously recorded in CA1. Diffuse activation of the medial entorhinal region was also obtained by repetitive low-intensity stimulation of the lateral olfactory tract at 2-8 Hz. Higher or lower stimulation frequencies did not induce hippocampal-medial entorhinal cortex activation. These results suggest that the medial and the lateral entorhinal regions have substantially different roles in processing olfactory sensory inputs.

Action Potentials↗

PubMed1999

Arterial supply of limbic structures in the guinea pig.

This study outlines the vascular territories of the cerebral arteries that originate from the Willis circle to supply limbic structures in the guinea pig brain. The entire cerebral vascular system was visualized in four preliminary experiments by performing superselective microangiographic studies with iodine contrast medium perfusion of the whole brain after in vitro isolation according to a technique described previously (de Curtis et al. [1991] Hippocampus 1:341-354). Subsequently, the perfusion territory of the different arteries that originate from the Willis circle was characterized after cannulation and perfusion of individual arteries with a gelatin solution that contained waterproof black ink. The analysis was performed by identifying the brain regions that contained the black stain on 150-microm-thick coronal sections that were cut after brain fixation with paraformaldehyde for at least 1 week. The middle cerebral artery and the rostral and caudal posterior cerebral arteries supply the limbic cortices and some related subcortical regions. In particular, large portions of the hippocampal formation are supplied by both the rostral posterior cerebral artery and the rostral branch of the caudal posterior cerebral artery, whereas the ventral temporal part of the hippocampus is served exclusively by the rostral posterior cerebral artery. The amygdala, the periamygdaloid cortex, and the piriform cortex are served by the middle cerebral artery and in part by the perforating arteries. The entorhinal, perirhinal, and postrhinal cortices are vascularized by the posterior and middle cerebral arteries, with a very broad overlap between the distal territories of these vessels. The demonstration of an extensive superimposition between the arterial supply of the entorhinal and the perirhinal regions suggests the presence of anastomotic connections that potentially are protective against ischemic events. Such an arrangement was not observed for the arteries that supply the ventral portion of the hippocampal formation and the basolateral amygdala, which showed nonoverlapping boundaries. The pathophysiological consequences of a similar vascular organization are discussed.

Animals↗

PubMed1999

Facilitation of spinal sciatic neuron responses to hindpaw thermal stimulation after formalin injection in rat tail.

A recent model of formalin injection in the tail induced a facilitation of the hindpaw withdrawal reflexes. In the present work we tried, after injecting formalin into the tail of the albino rat, to determine the spontaneous activity and response changes of lumbar sciatic wide-dynamic-range neurons to thermal stimulations of the paw at 45 degrees C and 48 degrees C (the respective thresholds for noxious and non-noxious thermal stimuli). The experiments were carried out with multiple recording electrodes placed in a comb array in the lumbar segments of the spinal cord at L4-L6 level in the sciatic projection field. A significant facilitation of the spontaneous activity was already evident 2 min after injection; at 5 min there were strong facilitations to the thermal stimuli. Stimuli at 45 degrees C, often ineffective prior to the formalin injection, became strongly excitatory. Stimuli at 48 degrees C evoked more conspicuous responses. This facilitatory effect on spontaneous and thermal responses followed a time-course comparable to that described for the excitations seen after paw formalin injection, but the duration was more prolonged, lasting more than 2 h. These data indicate a facilitatory role of the formalin effects on spinal sciatic neurons after injection in the tail. It is proposed that the mutual effects of spinal neurons in distant spinal segments could explain the facilitation and such a time-course, and that a role in the development of prolonged pain could be envisaged.

Animals↗

PubMed1999

Multifractal analysis of wide dynamic range neuron discharge profiles in normal rats and in rats with sciatic nerve constriction.

Using normal rats and rats with a chronic constriction injury of the sciatic nerve (injured, model of Bennett-Xie) we investigated the possibility of classifying, by statistical tools, the temporal sequences of neuronal discharges during different noxious and non-noxious stimuli. An analysis was made of both the distribution of the inter-spike intervals and the temporal density of spike trains, the latter being studied within the framework of stochastic universal multifractals, to allow the identification of different random processes involved in the discharge distributions through the Lévy index alpha. The statistical analysis shows that the parametrization based on the Lévy index seems able to discriminate between different noxious stimuli (mechanical pinching and thermal), both in normal and injured animals. Furthermore, comparing normal and injured animals, although the spontaneous basal and non-noxious stimuli (brushing) evoked activities presented different frequencies, these seem to have the same multifractal structure, while the corresponding statistics of the inter-spike intervals are quite different. This information might be relevant to the understanding of a code of neuronal firing and to the modelling of temporal patterns in acute and chronic noxious signals.

Algorithms↗

PubMed1998

Central effect of ketorolac involving NMDA receptors activity.

Possible central mechanisms underlying the analgesic action of Ketorolac, a non-steroidal antiinflammatory drug (NSAID) have been investigated using an iontophoretic approach. We found that the excitation induced by N-methyl-d-aspartate (NMDA) on spinal wide dynamic range (WDR) neurons was prevented, or reduced, by Ketorolac applied before or after the start of the NMDA ejection. The data suggest that Ketorolac can achieve its central analgesic effect by interfering with the NMDA receptor activity on the spinal neurons.

Animals↗

PubMed1998

Rough annealing by two-step clustering, with application to neuronal signals.

To accomplish analyses on the properties of neuronal populations it is mandatory that each unit activity is identified within the overall noise background and the other unit signals merged in the same trace. The problem, addressed as a clustering one, is particularly difficult as no assumption can be made on the prior data distribution. We propose an algorithm that achieves this goal by a two-phase agglomerative hierarchical clustering. First, an inflated estimation (overly) of the number of clusters is cast down and, by a maximum entropy principle (MEP) approach, is made to collapse towards an arrangement near natural ones. In the second step consecutive partitions are created by merging, two at time previously aggregated partitions, according to similarity criteria, in order to reveal a cluster solution. The procedure makes no assumptions about data distributions and guarantees high robustness with respect to noise. An application on real data out of multiple unit recordings from spinal cord neurons of mixed gas-anaesthetized rats is presented.

Algorithms↗