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L Sivilotti

Publications and source records attributed to L Sivilotti.

At least 19 recordsLinked to original sources
PubMed1997

A single serine residue confers tetrodotoxin insensitivity on the rat sensory-neuron-specific sodium channel SNS.

Sensory neurons express a sodium channel (SNS) that is highly resistant to block by tetrodotoxin (IC50 = 60 microM). SNS is 65% homologous to the cardiac sodium channel, in which a single hydrophilic residue in the SS2 segment is critical for tetrodotoxin resistance. By site-directed mutagenesis, we have substituted phenylalanine for serine at the equivalent position in SNS: this mutated (S356F) SNS channel is functionally similar to wild-type SNS when expressed in Xenopus oocytes, but is potently blocked by tetrodotoxin and saxitoxin with IC50s of 2.8 nM and 8.2 nM, respectively. These data provide clues to the rational design of selective blockers of SNS with potential as analgesic drugs.

Animals↗
PubMed1997

Structure and distribution of a broadly expressed atypical sodium channel.

A cDNA clone isolated from a rat dorsal root ganglion library encodes a 195 kDa voltage-gated sodium channel-like protein (SCL-11) with homology to the mouse (87%) and human (72%) atypical Na+ channels and rat partial clone NaG (98%). Two dominant mRNAs of 4.5 and 7 kb are expressed. The transcripts are present in lung, Schwann cells, pituitary and tissues containing smooth muscle cells. No functional channels could be detected on oocyte injection with cRNA, consistent with the absence of structural features necessary for voltage-gated sodium channel activity.

Amino Acid Sequence↗
PubMed1996

A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons.

Dorsal root ganglion sensory neurons associated with C-fibres, many of which are activated by tissue-damage, express an unusual voltage-gated sodium channel that is resistant to tetrodotoxin. We report here that we have identified a 1,957 amino-acid sodium channel in these cells that shows 65% identity with the rat cardiac tetrodotoxin-insensitive sodium channel, and is not expressed in other peripheral and central neurons, glia or non-neuronal tissues. In situ hybridization shows that the channel is expressed only by small-diameter sensory neurons in neonatal and adult dorsal root and trigeminal ganglia. The channel is resistant to tetrodotoxin when expressed in Xenopus oocytes. The electrophysiological and pharmacological properties of the expressed channel are similar to those described for the small-diameter sensory neuron tetrodotoxin-resistant sodium channels. As some noxious input into the spinal cord is resistant to tetrodotoxin, block of expression or function of such a C-fibre-restricted sodium channel may have a selective analgesic effect.

Amino Acid Sequence↗
PubMed1995

A P2X purinoceptor expressed by a subset of sensory neurons.

ATP is known to depolarize sensory neurons, and may play a role in nociceptor activation when released from damaged tissue. Here we report the molecular cloning and characterization of a new member of the P2X receptor family, P2X3, expressed by these cells. The channel transcript was present in a subset of rat dorsal-root-ganglion sensory neurons, some of which express nociceptor-associated markers; it was absent in other tissues that were tested, including sympathetic, enteric and central nervous system neurons. Moreover, when expressed in Xenopus oocytes, the channel showed an ATP-dependent cation flux. P2X3 is the only ligand-gated channel known to be expressed exclusively by a subset of sensory neurons. The remarkable selectivity of expression of the channel coupled with its sensory neuron-like pharmacology suggests that this channel may transduce ATP-evoked nociceptor activation.

Amino Acid Sequence↗
PubMed1994

The contribution of GABAA and glycine receptors to central sensitization: disinhibition and touch-evoked allodynia in the spinal cord.

1. Pain hypersensitivity is characterized by an increase in the response to noxious stimuli (hyperalgesia) and a reduction in threshold such that innocuous stimuli begin to elicit pain (allodynia). These sensitivity changes can be produced by an increase in excitability of dorsal horn neurons; the phenomenon of central sensitization. We have now examined whether a reduction in local segmental inhibitory mechanisms produces similar changes. The model system used for studying touch-evoked allodynia has been the recruitment of a low-threshold mechanoreceptor input to the nociceptive flexion withdrawal reflex in the decerebrate-spinal rat. 2. Hamstring flexor alpha motoneurons are characterized by high-threshold cutaneous receptive fields. Mechanical stimuli (pinch or firm pressure) evoke a brisk firing response in these cells, whereas low-intensity stimuli (light touch or brush) produce little or no effect, as expected for the output neurons of the nociceptive flexion withdrawal reflex. 3. Primary afferent C fiber conditioning inputs have previously been shown to produce prolonged increases in the excitability of the flexion reflex, as measured by the augmentation of the response to high-intensity peripheral stimuli. We have now examined whether these conditioning inputs and segmental disinhibition modify the responsiveness of the reflex to low-threshold inputs. 4. Brief (20 s), low-frequency (1 Hz), C fiber conditioning stimuli to the sural nerve increased the response of the hamstring flexor motor neurons to low-intensity cutaneous touch stimuli, reduced the cutaneous mechanical threshold, and increased the response to A beta inputs from the sural nerve. 5. Intrathecal injections of subconvulsant doses of the glycine receptor antagonist, strychnine (7 nmol) or the gamma-aminobutyric acid-A (GABAA) receptor antagonist, bicuculline (8 nmol) produced similar but longer lasting changes. The GABAB antagonist P-(3-aminopropyl)-P-diethoxymethyl-phosphonic acid (CGP 35348) had no significant effects. 6. The nociceptive flexion withdrawal reflex is under the control, therefore, of segmental inhibitory mechanisms mediated by glycine and GABAA receptors. Removal of this inhibition enables the reflex to be activated by low-intensity cutaneous stimuli. Given the similarities between the stimulus-response profiles of the nociceptive flexion reflex and the production of pain in man, these findings indicate that a decrease in the efficacy of spinal inhibitory circuits may contribute to the touch-evoked allodynia that occurs in pain hypersensitivity states, where A beta inputs begin to produce pain.

Afferent Pathways↗
PubMed1992

An intracellular study of the effects of GABA on frog tectal neurones in vitro.

The effects of gamma-aminobutyric acid (GABA) on neurones of the amphibian optic tectum were studied with current- and voltage-clamp recording from an isolated preparation of the midbrain of the frog Rana temporaria. Bath-applied GABA (1 mM) enhanced depolarizing synaptic potentials evoked in layer 6 tectal neurones by orthodromic stimulation of the optic tract. GABA also facilitated Na(+)- and Ca(2+)-dependent action potentials elicited by intracellular injection of depolarizing current. These actions of GABA were associated with comparatively small changes in membrane potential and their reversal potential was dependent on the Cl- equilibrium potential. Changes in input resistance observed during application of GABA were small and in part accountable for by the rectifying properties of the cell membrane. Tetrodotoxin (TTX; 1 microM) did not block the action of GABA on these neurones. These results show that externally applied GABA was able to raise directly the intrinsic excitability of frog tectal neurones and to enhance excitatory synaptic transmission elicited by stimulation of optic nerve fibres.

Action Potentials↗
PubMed1990

The effect of GABA on the frog optic tectum is sensitive to ammonium and to penicillin.

Excitatory postsynaptic potentials (termed U1 and U2) were extracellularly recorded from the frog optic tectum in vitro following electrical stimulation of the contralateral optic nerve. Gamma-Aminobutyric acid (GABA) and glycine elicited a large and sustained enhancement of these synaptic waves. In the presence of the Cl- transport inhibitor ammonium (NH+4) the effects of GABA or glycine were progressively reduced to about 50% of their initial action without changes in the control synaptic waves. In 50% Cl- media the depression of GABA and glycine responses by NH+4 was more intense. Other Cl- transport inhibitors such as bumetanide, piretanide and 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonate (SITS) were inactive against responses to GABA or glycine. Penicillin, a Cl- channel blocker, antagonized the action of GABA and glycine, while increasing the amplitude of the U2 waveform. The present results provide pharmacological evidence in support of the Cl- dependence of the unusual action of GABA or glycine in facilitating excitatory synaptic transmission in the optic tectum.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗
PubMed1990

An electrophysiological study of the action of N-methyl-D-aspartate on excitatory synaptic transmission in the optic tectum of the frog in vitro.

Excitatory synaptic field potentials, induced by stimulating optic nerve fibers, were recorded from in vitro preparations of the optic tectum of the frog. Bath-applied N-methyl-D-aspartate (NMDA), glutamate or quisqualate elicited transient enhancement in these field potentials, followed by a sustained depression reversible on washout. Responses to glutamate or quisqualate and the amplitude of control synaptic potentials, were not affected by the NMDA receptor antagonists aminophosphonovalerate (APV), 3(2-carboxypiperazin-4-yl)propyl-1-phosphonate (CPP), ketamine, magnesium ions or dizocipiline (MK 801) which, on the other hand, blocked the effects of NMDA. The antagonist dinitroquinoxaline-2,3-dione (DNQX), which is preferential for non-NMDA receptors, blocked the action of glutamate and synaptic transmission. In the presence of strychnine, glycine reversed the block of NMDA-mediated responses caused by magnesium. It is suggested that in the optic tectum of the frog, glutamate is the excitatory transmitter of at least one class of optic nerve fibers and that it acts through non-NMDA receptors. Although this area of the brain contains a well-developed NMDA receptor system, its function in physiological synaptic transmission remains to be elucidated.

Animals↗
PubMed1989

Pharmacology of a novel effect of gamma-aminobutyric acid on the frog optic tectum in vitro.

Excitatory postsynaptic field potentials (termed U1 and U2 waves) were extracellularly recorded from superficial layers of the frog optic tectum in vitro following stimulation of the contralateral optic nerve. gamma-Aminobutyric acid (GABA) largely enhanced the U1 and U2 waves, an effect which was progressively diminished by lowering the extracellular Cl- content. Muscimol or 3-aminopropanesulphonic acid (3-APS) was more potent than GABA in enhancing the U1 and U2 waveforms, whereas trans-4-aminocrotonic acid (TACA) was equipotent, its cis isomer (CACA) being less active. The action of GABA, TACA and CACA was relatively insensitive to bicuculline (100 microM). Pentobarbitone increased the U1 and U2 waves and antagonized the action of GABA. Midazolam did not change the GABA-induced responses. Picrotoxin antagonized both pentobarbitone and GABA effects. It is suggested that in the frog optic tectum GABA can activate an atypical receptor mechanism blocked by picrotoxin but comparatively insensitive to bicuculline.

Animals↗
PubMed1989

Antagonism of the actions of glutamate by pentobarbitone or midazolam in the frog optic tectum in vitro.

Excitatory synaptic transmission, induced by electrical stimulation of optic nerve fibres on relay neurones, was recorded from in vitro preparations of the optic tectum of the frog. Bath-applied glutamate (the putative excitatory transmitter of the optic nerve) produced transient enhancement of tectal field potentials, followed by a depression, presumably caused by sustained neuronal depolarization. Pentobarbitone potently antagonized the depressant effect of glutamate, producing an approximate 50% reduction in the response of the tectum to glutamate at 25 microM. Midazolam also decreased the effect of glutamate with an IC50 value of 5 nM. Since, in the optic tectum of the frog, neither pentobarbitone nor midazolam enhance responses to bath-applied GABA, it is suggested that this area of the brain is a useful preparation in which to investigate the interaction of barbiturates and benzodiazepines with glutamate receptor mechanisms, without concurrent interactions with GABAergic processes.

Animals↗
PubMed1988

Nipecotic acid enhances synaptic transmission in the frog optic tectum by an action independent of GABA uptake inhibition.

In the frog optic tectum maintained in vitro at 7 degrees C to depress gamma-aminobutyric acid (GABA) transport systems the GABA uptake inhibitor nipecotic acid reversibly enhanced field potentials evoked by optic nerve stimulation. This effect was dose dependent and picrotoxin sensitive. Responses to low concentrations of GABA agonists were not affected by nipecotic acid, whereas responses to the maximal muscimol concentration were depressed. It is suggested that in addition to its well known inhibitory action on GABA uptake systems, nipecotic acid can evoke GABA mimetic effects whose precise origin is presently unclear.

Animals↗
PubMed1988

Complex effects of baclofen on synaptic transmission of the frog optic tectum in vitro.

In the frog optic tectum in vitro, low concentrations of the GABAB agonist (-)-baclofen reversibly depressed field potentials evoked by optic nerve stimulation. This effect, which was dose-dependent and stereo-specific, differed from the potent facilitatory action of bath-applied GABA. A transient enhancement in field potential amplitude often preceded the more consistently observed depression. Both phases of the baclofen response were insensitive to the GABAA antagonist picrotoxin. Since the effects of baclofen and GABA were different, it is suggested that GABAB receptors were unlikely to mediate the action of GABA on synaptic transmission in the optic tectum.

Action Potentials↗
PubMed1986

Biphasic effects of glycine on synaptic responses of the frog optic tectum in vitro.

Electrophysiological recordings from the in vitro amphibian optic tectum were used to study the action of bath-applied glycine (Gly) on synaptic transmission between optic tract fibres and tectal neurones. Gly enhanced synaptic field potentials (ED50 = 0.3 mM) and, when applied in concentrations greater than or equal to 1 mM subsequently blocked them. All these effects of Gly were antagonized by 0.2-1 microM strychnine. Recordings from optic tract afferent fibres indicated that Gly increased the amplitude of small submaximal compound action potentials while depressing those which were near maximal. It is suggested that Gly exerted a novel modulatory effect on synaptic transmission via a combination of pre- and postsynaptic sites of action.

Action Potentials↗
PubMed1986

Field potentials from the frog optic tectum in vitro.

A new method for maintaining the isolated frog optic tectum superfused in vitro at a low temperature is described. The preparation remains viable for several hours with electrophysiological responses similar to those of the in vivo tectum. This superfusion technique will allow controlled application of putative neurotransmitters and other substances in known concentrations and therefore makes the in vitro frog optic tectum a useful model system for neuropharmacological investigations into central optic pathways.

Animals↗
PubMed1985

The effect of nicotine and cytisine on 3H-acetylcholine release from cortical slices of guinea-pig brain.

Nicotine 1.8 X 10(-5)-1.8 X 10(-4) mol/l enhanced the spontaneous 3H-efflux from guinea-pig cortical slices preloaded with 3H-choline and perfused in the presence of hemicholinium (HC-3). The facilitation of tritium outflow was prevented by tetrodotoxin 5 X 10(-7) mol/l and by D-tubocurarine 4.5 X 10(-6) mol/l. Nicotine 1.8 X 10(-6)-1.8 X 10(-4) mol/l, and the agonist cytisine 5 X 10(-7)-5 X 10(-5) mol/l increased, in a concentration-dependent way, 3H-efflux from electrically-stimulated slices (0.2 Hz). The concentration-response curves of both drugs were parallelly shifted to the right by D-tubocurarine 4.5 X 10(-6) mol/l. The EC50 values (i.e. the concentrations required to cause a 50% increase in the S2/S1 ratio) changed for nicotine from 5.58 X 10(-5) to 4.34 X 10(-4) and for cytisine from 6.3 X 10(-6) to 2.75 X 10(-4) mol/l in the absence and in the presence of the antagonist, respectively. In the range of 0.2-2 Hz the magnitude of the effect of nicotine was inversely related to the rate of stimulation. The response to nicotine was subject to rapidly developing tachyphylaxis; it was resistant to atropine. It is concluded that nicotine and cytisine facilitate 3H-efflux from the cholinergic nerve endings of guinea-pig cerebral cortex. This effect involves sodium-dependent mechanisms and is due to an interaction of the drugs with receptors showing affinity for D-tubocurarine.

Acetylcholine↗
PubMed1985

Pharmacological characterization of D-aminophosphonovaleric acid antagonism of amino acid and synaptically evoked excitations on frog motoneurones in vitro: an intracellular study.

The effect of D-aminophosphonovaleric acid (D-APV) on the depolarizations induced by N-methyl-D-aspartate (NMDA), glutamate, aspartate or quisqualate was studied with intracellular recordings from frog motoneurones in vitro. D-APV (0.5-10 microM) produced a slight hyperpolarization of the motoneuronal membrane without significant changes in input conductance. In control and tetrodotoxin-containing solutions the depolarizations induced by NMDA were strongly reduced by D-APV while quisqualate depolarizations were unaffected. Responses to glutamate and aspartate were antagonized to an intermediate level. The relatively small conductance increases evoked by excitatory amino acids were unaltered in solutions containing D-APV. The amplitude of monosynaptic excitatory postsynaptic potentials (e.p.s.ps) was strongly depressed by D-APV. The amplitude of polysynaptic e.p.s.ps was little changed but their decay time was reduced. It is suggested that D-APV is a powerful and selective NMDA receptor antagonist and that an endogenous amino acid acting via NMDA receptors may be the transmitter of monosynaptic e.p.s.ps on frog motoneurones.

2-Amino-5-phosphonovalerate↗