Properties of antidromically identified neurons in the enkephalinergic magnocellular dorsal nucleus of the guinea pig hypothalamus

In the guinea pig, immunocytochemical and neuroanatomical studies have demonstrated that enkephalin-containing neurons in the hypothalamic magnocellular dorsal nucleus (MDN) terminate in the lateral septum (LS). In the present investigation, 114 MDN neurons, studied with extracellular recording techniques, were identified by antidromic activation from the LS. Latencies of responses from ipsilateral and contralateral LS were 13.5 and 18.78 ms, respectively, corresponding to an axonal conduction velocity of 0.1 m/s. By using the reciprocal collision test, evidence is presented for bilateral projection of individual MDN neurons to the LS. Fifty-one (44.73%) MDN-LS neurons discharged in a slow irregular pattern. Interspike time histograms were very similar and had a mode of about 280 ms. Peristimulus time histograms were compiled from 15 active MDN-LS neurons. Stimulation which elicited antidromic spikes resulted in a brief silent period in the spontaneous activity which was related to the normal interspike interval pattern of the firing. Prolonged silent periods as well as silent period occurring after subthreshold stimulus and increasing with the stimulus intensity were attributed to inhibitory synaptic effects. On the other hand, some MDN-LS neurons displayed orthodromic excitatory responses following LS stimulation. These observations provide electrophysiological evidence of a direct MDN-LS pathway, in all likelihood of enkephalinergic nature, and indicate that some MDN-LS neurons receive inhibitory and excitatory afferents from the LS.     

Postnatal development of alpha-adrenoceptor-mediated autoinhibition in the locus coeruleus

Rats, from birth to postnatal day 34, were anesthetized with urethane and a neuropharmacological study was carried out of the autoreceptors located on the somadendritic membranes of locus coeruleus (LC) neurons. Iontophoretic application of noradrenaline (NA) caused inhibition of LC cell firing at all developmental stages, and such inhibition was totally blocked by the alpha 2-antagonist piperoxane. The sensitivity of LC neurons to iontophoretically applied NA appeared to become reduced with age. In LC neurons from birth to postnatal day (PD) 8, the prolonged period of suppressed firing after antidromic activation by stimulation of the dorsal noradrenergic bundle was not shortened by piperoxane. After PD 9, the proportion of LC neurons in which piperoxane could antagonize the postactivation inhibition increased with age. These results indicated that although LC neurons, even at birth, had alpha 2-adrenoceptors on the somadendritic membranes which were responsible for the NA-induced inhibition, inhibition of LC cell firing caused by NA released from the terminals of axon collaterals and/or possibly from dendrodendritic synapses did not occur until PD 9.     

Concerning recurrent collaterals and afferent fibers in the hypoglossal nerve of the rat

The pause in firing of hypoglossal motoneurons following a stimulus to the hypoglossal nerve in the rat could be explained by an afterhyperpolarization or by an inhibition via recurrent collaterals or afferent fibers. Extracellular and intracellular recordings from glass microelectrodes have established the all-or-none nature of both the pause in electrophoretically induced firing and a hyperpolarization which followed antidromic invasion. Only stimulation of the hypoglossal nerve branch which evoked antidromic invasion resulted in the hyperpolarization. The threshold for these two events was identical. Stimulating the hypoglossal nerve branches over a whole range of stimulus strengths produced no other potential changes, neither were any neurons with Renshaw-cell characteristics found. No evidence for the presence of afferent fibers in the hypoglossal nerve was abtained from recording the compound action potential following distal stimulation, from examination of the appropriate segment by electron microscopy, or from a search for reflex effects in other cranial nerves and on blood pressure following stimulation of the hypoglossal nerve. Peaks of increased probability in firing following the pause seen in poststimulus histograms were attributed to synchronization between the computer sweep and the time course of the afterpotential following antidromic invasion.     

Electrophysiological identification of neurons in locus coeruleus

In 17 urethane-anesthetized rats 35 neurons, histologically verified as being situated in the locus coeruleus, were driven antidromically (latency, 44 msec) by electrical stimulation of the supracallosal bundle. Neurons of the locus coeruleus were also activated antidromically by stimulation of sites along the dorsal noradrenergic bundle in the midbrain (8-msec latency) and the hypothalamus (12-msec latency), and by stimulation of sites in the olfactory bulb (latency, 39 msec). Conduction velocity from these sites to the locus coeruleus was estimated to be 0.4 to 0.6 m/sec. Refractory periods of fibers in the dorsal noradrenergic bundle were determined at twice threshold and in the supracallosal bundle at intensities just above threshold; refractory periods ranging from 4 to 20 msec were observed. Because neurons both in and near the locus coeruleus were antidromically activated by stimulation of the dorsal noradrenergic bundle whereas stimulation of the supracallosal bundle antidromically activated only neurons in the locus coeruleus, stimulation of the dorsal noradrenergic bundle could not be used to identify locus coeruleus neurons. It is concluded that a subpopulation of neurons in the locus coeruleus can be identified by their slow, steady firing rate (2.6 per second) and long-latency antidromic response to stimulation of the supracallosal bundle. The electrophysiological properties of locus coeruleus neurons are considered in relation to neuroanatomical and functional studies of the locus coeruleus.     

Locus coeruleus activity in vitro: intrinsic regulation by a calcium-dependent potassium conductance but not alpha 2-adrenoceptors

Locus coeruleus neurons recorded intracellularly in rat brainstem slices exhibited spontaneous activity and a marked afterhyperpolarization following a burst of spikes. This afterhyperpolarization was associated with an increase in membrane conductance and resulted in a marked postactivation inhibition of spontaneous activity. Since the reversal potential of the afterhyperpolarization was found to be virtually identical when recorded with KCl or potassium acetate-filled electrodes and shifted in the hyperpolarizing and depolarizing direction with decreases and increases in extracellular potassium concentrations, respectively, the afterhyperpolarization seen following a burst of spikes in this cell group appears to be mediated by an increase in potassium conductance. The afterhyperpolarization and postactivation inhibition were markedly attenuated by reducing calcium influx by either omitting extracellular calcium in the bathing medium or blocking calcium channels with manganese or cadmium. Thus, the afterhyperpolarization and the resulting postactivation inhibition appear to be largely mediated by the activation of a calcium-dependent potassium conductance. Previous reports in vivo have suggested that activation of alpha 2-adrenoceptors by norepinephrine release from recurrent axons or dendrites may mediate self-inhibition in the locus coeruleus. In this study, we examined the effect of blocking alpha 2-adrenoceptors on the afterhyperpolarization and postactivation inhibition. Administration of the alpha 2-adrenoceptor antagonist piperoxane failed to produce any changes in either of these parameters, suggesting that at least in vitro the afterhyperpolarization and postactivation inhibition seen in locus coeruleus neurons do not result from the activation of alpha 2-adrenoceptors.     

Pyramidal neurons in rat prefrontal cortex show a complex synaptic response to single electrical stimulation of the locus coeruleus region: Evidence for antidromic activation and GABAergic inhibition using in vivo intracellular recording and electron microscopy

Cognition and acquisition of novel motor skills and responses to emotional stimuli are thought to involve complex networking between pyramidal and local GABAergic neurons in the prefrontal cortex. There is increasing evidence for the involvement of cortical norepinephrine (NE) deriving from the nucleus locus coeruleus (LC) in these processes, with possible reciprocal influence via descending projections from the prefrontal cortex to the region of the LC. We used in vivo intracellular recording in rat prefrontal cortex to determine the synaptic responses of individual neurons to single electrical stimulation of the mesencephalic region including the nucleus LC. The most common response consisted of a late-IPSP alone or preceded by an EPSP. The presence of an early-IPSP following the EPSP was sometimes detected. Analysis of the voltage dependence revealed that the late-IPSP and early-IPSP were putative K⁺- and Cl⁻ dependent, respectively. Synaptic events occurred following short delays and were inconsistent with the previously reported time for electrical activation of unmyelinated LC fibers. Moreover, systemic injection of the adrenergic antagonists propranolol (β receptors), or prazosin (α1 receptors), did not block synaptic responses to stimulation of the LC region. Finally, certain neurons were antidromically activated following electrical stimulation of this region of the dorsal pontine tegmentum. Taken together, these results suggest that the complex synaptic events in pyramidal neurons of the prefrontal cortex that are elicited by single electrical stimulation of the LC area are mainly due to antidromic activation of cortical efferents. Further insight into the chemical circuitry underlying these complex synaptic responses was provided by electron microscopic immunocytochemical analysis of the relations between the physiologically characterized neurons and either 1) GABA or 2) dopamine-β-hydroxylase (DBH), a marker for noradrenergic terminals. GABA-immunoreactive terminals formed numerous direct symmetric synapses on somata and dendrites of pyramidal cells recorded and filled with lucifer yellow (LY). In contrast, in single sections, noradrenergic terminals immunoreactive for DBH rarely contacted LY-filled somata and dendrites. These results support the conclusion that IPSPs observed following single electrical stimulation of the LC region are mediated bu GABA, with little involvement of NE. These IPSPs, arising from antidromic invasion of mPFC cells innervating the LC, may improve the signal-to-noise ratio and favor a better responsiveness of neighboring neurons to NE released in the mPFC. © 1996 Wiley-Liss, Inc.     

Evidence for self- and neighbor-mediated postactivation inhibition of locus coeruleus neurons

Activation of locus coeruleus (LC) neurons is typically followed by inhibition of impulse activity lasting hundreds of ms. Previous studies have implicated two possible mechanisms for this postactivation inhibition: collateral synaptic interactions among LC neurons; and spike-induced, calcium-activated potassium conductance in the soma-dendritic membrane of LC cells. In the present study, antidromic or sensory stimuli were presented at near-threshold intensities for activation of LC neurons. A special computer program accumulated activity for trials yielding driven responses separately from that for trials of identical stimuli during the same train that failed to evoke activity. We found significant inhibition of LC impulse activity for antidromic or sensory stimuli that failed to excite the recorded cell as well as for stimuli that activated the recorded cell. The former result precludes an essential role of intrinsic inhibitory membrane currents (e.g. calcium-activated potassium conductance) in generating postactivation inhibition. Administration of the alpha antagonist piperoxane reduced the magnitude of inhibition on both driven and non-driven trials. Our findings indicate that inhibition on non-driven trials appears to be a synaptically mediated phenomenon, perhaps reflecting norepinephrine released from neighboring LC neurons that are activated. Furthermore, our data support the presence of a spike-dependent mechanism that also contributes substantially to postactivation inhibition in these cells. Thus, the overall results indicate the presence of two intracoerulear mechanisms that mediate postactivation inhibition characteristic of noradrenergic LC neurons.     

Neurophysiological consequences of presynaptic receptor activation: changes in noradrenergic terminal excitability

Experiments were carried out to explore the view that activation of presynaptic receptors on the terminals of noradrenergic neurons is accompanied by alterations in their excitability to direct electrical stimulation. Antidromic action potentials evoked from frontal cortex of urethane anesthetized rats were recorded extracellularly from nucleus locus coeruleus. The threshold current necessary to evoke antidromic action potentials varied as a result of infusion of adrenergic agonists and antagonists into frontal cortex within 50 micrometer of the stimulating electrode. Local infusion of the alpha-adrenergic agonist clonidine produced a marked decrease in terminal excitability, while the alpha-antagonist phentolamine produced an increase in terminal excitability and was shown to reverse the effect of the agonist. Infusion of the beta-adrenergic agonist isoproterenol was without effect, although the beta-antagonist propranolol resulted in a decrease in terminal excitability. Infusions of potassium increased excitability of locus coeruleus terminals. Terminal excitability was seen to vary inversely with the rate of spontaneous or high frequency stimulation-induced firing of locus coeruleus neurons. From these observations, it may be inferred that activation or blockade of alpha-adrenergic presynaptic receptors results in changes in polarization and/or conductance of the noradrenergic synaptic endings. These results are discussed with respect to phenomena associated with the possible presynaptic regulation of neurotransmitter release.     

Recurrent inhibition of firing motoneurones in man

The effects of antidromic stimulation of large motoneurones on firing small motoneurones of soleus muscle have been studied. Against the background of rhythmic firing of small motor units (MUs) activated during weak voluntary muscle contraction, thick efferents of the tibialis posterior nerve were selectively stimulated and an M response was evoked in which small MUs were not involved. This provided a means of avoiding antidromic stimulation of the motoneurone under study and, thus, analysing the effect of stimulation without its summation with after-hyperpolarization. The background firing rate of MUs was 4.5-9.2/sec. PSTHs revealed a distinct inhibitory effect with a latency of 35-40 msec (slightly exceeding the latency of monosynaptic reflex) and duration 5-30 msec. It was concluded that the short-latency inhibition could be identified as recurrent inhibition. The effectiveness of recurrent inhibition evaluated by the lengthening of the interspike interval was shown to depend on the arrival time of the volley in the interval and on the background firing rate of the motoneurone. When the inhibitory volley arrived at the beginning of the interspike interval it was ineffective. This indicates that in the investigated range of firing rates the motoneurone is unable to exert an inhibitory effect on its own firing via recurrent collaterals. The inhibitory volley became highly effective at the end of an interspike interval, when the membrane potential approached threshold. The lengthening of interspike interval was more marked at a lower firing rate of the motoneurone. An increase in the background firing rate reduced the extent of recurrent inhibition (at a rate above 10/sec up to its complete ineffectiveness).(ABSTRACT TRUNCATED AT 250 WORDS)     

An automatic device for determining threshold variations in antidromically activated neurons

A device was designed and constructed with the purpose of evaluating threshold variations for antidromic invasion of extracellularly recorded neurons. Identification of a neuron is carried out by two procedures, an amplitude discriminator, which isolates the spike from the baseline noise, and by a latency window which is set accordingly to the neuron's antidromic latency. During threshold evaluation, the duration of an electric pulse applied to the neuron's axon is automatically varied depending on the presence or not of an action potential. For a given spike, the stimulus is progressively decreased (-delta i) up to a point where the neuron ceases to respond and thereafter, the stimulus amplitude is progressively increased (+delta i) until slightly suprathreshold values are obtained. The procedure guarantees a discharge probability of the neuron equivalent to 50% of all applied stimuli, and the simple monitoring of the stimulus amplitude is enough to obtain the threshold value for a predetermined intensity. The reliability of this device was checked in studies related to threshold variations in neurons antidromically driven in prefrontal cortex following stimulation of the ipsi and contralateral olfactory bulb. Variations in excitability were found during and following tetanic stimulation and throughout the axon's supernormal conduction period. This technique allows the assessment of threshold variations in antidromic driving, not only in the present experimental design, but also in other conditions induced by changes in extracellular ionic concentrations, drug applications or in those produced by excitatory or inhibitory synaptic activity on the neuron under study.     

Amphetamine''s effects on terminal excitability of noradrenergic locus coeruleus neurons are impulse-dependent at low but not high doses

The actions of amphetamine in the locus coeruleus and its terminal fields in the frontal cortex were studied using extracellular recording to measure terminal excitability, firing rate and the probability of antidromic action potential invasion of the somatodendritic region in urethane anesthetized rats. At low dose (0.25 mg/kg), amphetamine increased terminal excitability. In comparison, subsequent administration of the highest dose (5.0 mg/kg, i.v.) of amphetamine tested suppressed neuronal firing and blocked antidromic action potential invasion of the somatodendritic region. Despite the absence of impulse traffic, high dose amphetamine reversed the effect of low dose amphetamine in the terminal field and decreased terminal excitability. The alpha 2 antagonist, yohimbine (0.5 mg/kg, i.v.), reversed the effects of high dose amphetamine on terminal excitability and somatodendritic invasion without reinstating neuronal firing. Noradrenergic autoreceptor agonists are known to decrease terminal excitability, whereas antagonists are known to increase terminal excitability. Thus, since low dose amphetamine produces the same effect on terminal excitability that antagonists do, it appears that low dose amphetamine may reduce autoreceptor activation by reducing norepinephrine release in frontal cortex as a consequence of inhibiting locus coeruleus neuronal firing. In contrast, high dose amphetamine acts like autoreceptor agonists do and decreased terminal excitability. Hence high dose amphetamine may increase norepinephrine release, even in the absence of impulse traffic.     

Evidence for functional contact between cografted locus coeruleus and spinal cord in oculo: electrophysiological studies

The functional consequences of the locus coeruleus innervation of the spinal cord are not yet clearly understood. In a recent histological study it was shown that intraocular spinal cord grafts will become innervated by tyrosine hydroxylase-positive nerve fibers from a cografted locus coeruleus. In the present study we use this intraocular model of the descending coeruleo-spinal pathway to investigate functional contact between locus coeruleus and the spinal cord. We have pharmacologically characterized the receptor mediation of norepinephrine-induced, as well as locus coeruleus-mediated depressions of spinal cord neurons grafted in oculo. We found that electrical stimulation of the locus coeruleus part of the double grafts predominantly caused an inhibition of cografted spinal cord neurons. Norepinephrine-induced inhibition of the firing rate of single grafted spinal cord neurons was antagonized by phentolamine, an alpha-adrenergic antagonist, but was unaffected by timolol, a beta-adrenergic antagonist. Similarly, inhibition of the firing rate of grafted spinal cord neurons by stimulation of cografted locus coeruleus was antagonized by phentolamine but not by timolol. Interestingly, single spinal cord grafts were more sensitive to the depressant effects of perfused norepinephrine than was the spinal cord cografted with locus coeruleus. We conclude that spinal cord grafts can be functionally innervated by cografted locus coeruleus and that the noradrenergic inputs to spinal cord from cografted locus coeruleus are alpha-adrenergically mediated. Furthermore, the postsynaptic receptors in single spinal cord grafts appear to be supersensitive to norepinephrine application.     

Dorsal raphe stimulation produces inhibitory effect on trigeminal nucleus neurons.

Inhibitory effects of conditioning stimulation of the dorsal raphe nucleus (DR) on the neuron activity in the rostral part of spinal trigeminal nucleus (STN) were studied in cats for the purpose of comparison with the inhibition induced by locus coeruleus (LC) stimulation. DR conditioning stimulation reduced the orthodromic field potential in STN elicited by inferior alveolar nerve stimulation, and enhanced the antidromic field potential in the trigeminal nerve evoked by STN stimulation; but the inhibitory effects of DR stimulation were considerably weaker than those of LC stimulation. In tracking experiments near the raphe nucleus, conditioning stimulation of DR itself produced the most pronounced decrease in the STN field potential. Orthodromic spike number of STN relay neurons was significantly reduced by DR conditioning stimulation; however, the threshold for the conditioning stimulus to the DR was much higher than that to the LC. Antidromic spike generation of the STN neurons was unaltered by conditioning stimulation of both DR and LC. DR stimulation elicited a field potential in STN, which followed high frequency stimuli up to 200 HZ. A single fiber action potential was also obtained in STN by DR stimulation. STN stimulation produced a field potential in DR, which followed high frequency stimuli. It is suggested from these findings that conditioning stimulation of DR produces a direct inhibition of transmission in STN neurons; however, this stimulation has less effect on these neurons than does stimulation of the LC.     

Noradrenergic hyperinnervation of the trigeminal sensory nuclei

Administration of 6-hydroxydopamine to neonatal rats results in a permanent increase in the norepinephrine content in several brainstem areas. To assess the physiological effects of this hyperinnervation, we studied the noradrenergic inhibition of transmission of sensory information through the principal sensory and rostral spinal trigeminal nuclei. Unit activity produced by tactile stimulation of the face was recorded extracellularly from trigeminal sensory neurons in normal and hyperinnervated rats. The noradrenergic neurons projecting to the trigeminal sensory nuclei (locus coeruleus and the region of the lateral lemniscus) were stimulated 40 ms prior to delivery of a tactile stimulus to the face, producing complete inhibition. The interstimulus interval was then increased in 100 ms increments until the sensory response returned to control values. Compared with controls, the duration of inhibition was 30% longer in hyperinnervated rats and 25% shorter in rats depleted of catecholamines with reserpine and alpha-methyl-p-tyrosine. While the beta-adrenergic blocker, propranolol, had no effect on the duration of inhibition in normal animals, the mean latency of response to tactile stimulation was decreased from 15.3 to 10.4 ms. Propranolol given to hyperinnervated rats decreased the latency of the response to tactile stimulation from 15.1 to 9.1 ms and decreased the duration of inhibition by 40% compared with untreated hyperinnervated rats, suggesting an alteration in numbers or sensitivity of beta-receptors. Since the drug treatment never eliminated the inhibition due to locus coeruleus stimulation, there is also a non-noradrenergic component. We conclude from these observations that noradrenergic hyperinnervation is not completely counteracted by receptor down regulation.     

The role of locus coeruleus in the antiepileptic activity induced by vagus nerve stimulation

Stimulation of the vagus nerve produces antiepileptic effects. This is used clinically to treat drug-refractory epilepsies. The mechanisms responsible for these effects depend on the activation of vagal afferents reaching the nucleus of the solitary tract. This review focuses on the neuroanatomy of the nucleus of the solitary tract and its relation with the nucleus locus coeruleus as a preferential anatomical substrate in producing antiepileptic effects. In fact, following the transient or permanent inactivation of locus coeruleus neurons, some antiepileptic effects of vagus nerve stimulation are lost. The activation of locus coeruleus per se is known to limit the spread of a seizure and the duration of a variety of seizure types. This is due to the fine chemical neuroanatomy of norepinephrine pathways that arise from the locus coeruleus, which produce widespread changes in cortical areas. These changes may be sustained by norepinephrine alone, or in combination with its co-transmitters. In addition, vagus nerve stimulation may prevent seizures by activating the serotonin-containing dorsal raphe neurons.     

Electrophysiological evidence for axonal degeneration of locus coeruleus neurons following long-term forced running stress

Using electrophysiological methods, a change in the density of axon terminals of locus coeruleus (LC) neurons in the cerebral cortex of rats following long-term forced running stress was examined. The stressed animals were classified into two groups based on spontaneous running activity (SRA) measured for 2 weeks after the stress treatment: 1) animals showing early restoration of SRA (poststress active rat) and 2) animals showing little or no SRA (poststress inactive rat). To quantify the density of LC axon terminals in the cerebral cortex, the percentage of LC neurons antidromically activated by cortical stimulation (projection index, P-index) was assessed. The P-indices for the cortex decreased in the poststress inactive rats. Since the threshold currents for antidromic activation were not altered by the stress treatment, the observed change was considered to reflect a change of the density of LC axon terminals rather than physiological consequences. Therefore, when animals receive a prolonged, severe stress, LC neurons in a certain group of the animals may cause axonal retraction or degeneration in the cerebral cortex.     

Decrease in the Reactivity of Locus Coeruleus Neurons to Hypotension after an Increase in their Tyrosine Hydroxylase Content: A Subregional In Vivo Voltammetry Study in the Rat

The aim of the present work was to determine if noradrenergic neurons of the anterior and the posterior subregions of the locus coeruleus exhibit a difference in reactivity in response to sodium nitroprusside-induced arterial hypotension, and if the pharmacological induction of tyrosine hydroxylase by RU24722 modifies the reactivity of locus coeruleus neurons to this hypotensive stimulus. Previous findings have demonstrated that administration of RU24722 increases the concentration of tyrosine hydroxylase in the rat locus coeruleus by two different mechanisms in the anterior and in the posterior locus coeruleus subregions. The goal of the present study was to measure in vivo the changes in catecholaminergic metabolism in the locus coeruleus after treatment with RU24722 using differential normal pulse voltammetry (DNPV). In vehicle-treated rats, arterial hypotension increased catecholaminergic metabolism with the same pattern in the two locus coeruleus subregions. However, the changes in the magnitude of the catechol oxidation current throughout the recording period were significantly smaller in the posterior subregion ( P < 0.001). In the RU24722-pretreated rats, there was a 39% increase in tyrosine hydroxylase and dihydroxyphenylacetic acid in the locus coeruleus. The functional reactivity to hypotension measured by DNPV was significantly decreased ( P < 0.001) in both the anterior and posterior locus coeruleus subregions with RU24722 treatment. Therefore, this study suggests that the response of locus coeruleus cells to a hypotensive stimulus depends upon the intracellular tyrosine hydroxylase concentration both in the basal condition and during pharmacological induction of tyrosine hydroxylase gene expression.     

Two physiologically distinct populations of neurons in the ventrolateral medulla innervate the locus coeruleus

Recent anatomic studies indicate that the nucleus paragigantocellularis (PGi), located in the rostral ventrolateral medulla, strongly innervates the locus coeruleus (LC) while no such input derives from the more caudally located lateral reticular nucleus (LRN). In the present study, focal electrical stimulation of the LC was used to antidromically activate neurons in the ventrolateral medulla. A substantial number of PGi neurons were antidromically driven from the ipsilateral LC, while antidromic activation was virtually absent in LRN. Furthermore, several physiologic properties of antidromically driven cells in PGi define two populations within this group of neurons afferent to LC. These findings provide physiologic confirmation of an anatomically identified input to LC.     

Australian funnel-web spider toxin, versutoxin, enhances spontaneous synaptic activity in single brain neurons in vitro

Neurotoxins isolated from the venoms of Australian funnel-web spiders increase spontaneous action potential activity in a variety of excitable cells. In the present study intracellular recordings were made with microelectrodes (30–60 MΩ, 2 M KCl) from locus coeruleus, mesencephalic nucleus of the trigeminal nerve and laterodorsal tegmental neurons in brain slices. Versutoxin, a polypeptide toxin isolated from the venom ofHadronyche versutus produced a profound increase in spontaneous synaptic activity impinging on neurons, which did not fully recover for up to 3 h after washout. The threshold concentration was 1.5 nM in locus coeruleus neurons, with increasing concentrations (up to 50 nM) producing larger effects. A modest increase in synaptic activity was observed in mesencephalic nucleus of the trigeminal nerve neurons during superfusion with 50 nM versutoxin. The increase in spontaneous synaptic activity was reversed by agents which block synaptic potentials impinging on locus coeruleus neurons, i.e., tetrodotoxin (100 nM), Co²⁺ (3 nM) or the combination of 6-cyano-7-nitroquinoxaline-2,3-dione (10 μM) and bicuculline (30 μM). Threshold, peak amplitude, maximum rate of rise, duration, amplitude of afterhyperpolarisations and interspike intervals of action potentials in each type of neuron were unaffected by versutoxin. Voltage-current relationships were also unaffected. Calcium-dependent action potentials evoked in locus coeruleus neurons in the presence of tetrodotoxin were unaffected by versutoxin, as were depolarisations produced by exogenously applied glutamate. These results suggest that versutoxin increases spontaneous synaptic activity, but has no effect on the membrane properties of the soma of several types of rat brain neurons.     

Projection of tooth pulp afferents to the cat trigeminal nucleus caudalis

Unit activity was recorded extracellularly from cat medullary neurons following electrical stimulation of the canine tooth pulp. Response characteristics of the neurons quickly stabilized at specific suprathreshold stimulus intensities but such properties as spike latency, interspike interval and spike density varied systematically as intensity was raised to maximally effective values. Receptive fields were principally unilateral. The majority included both canines and extended into other oro-facial areas. Suppression of a pulpal response could be effected by preceding tooth stimulation with a conditioning stimulus applied to some other point in the receptive field of the responding cell at an appropriate interstimulus interval. In contrast, a pulpal response could be enhanced by presenting two stimuli successively to the same canine at such intervals. Similar enhancing effects followed simultaneous stimulation of spatially segregated loci in a field. The pulp-responsive neurons were localized histologically in, or in the immediate vicinity of, the nucleus caudalis of the spinal trigeminal complex where the possibility of their existence has been questioned previously. Most of the cells were situated along the ventromedial border of the nucleus, a region reported to contain other pain-related neurons with trigeminal fields.