G. Li Volsi

Effects of 5-hydroxytryptamine on the firing rates of neurons of the lateral vestibular nucleus in the rat

Summary5-hydroxytryptamine (5-HT) was delivered microiontophoretically (20–80 nA) to cells of the lateral vestibular nucleus of anaesthetized rats to test its influence on the spontaneous activity of single neurons. 5-HT increased the rate of firing of 94% of the units tested. The enhancement persisted for up to 700 s after the end of the 5-HT ejection and the maximum magnitude of the excitation (10–3400%) showed a hyperbolic correlation (ϱ=0.86) with background firing. In 43% of units the enhancement was preceded by a short-lasting (less than 105 s) depression of the neuronal firing rate, the magnitude of which was unrelated to the background mean firing rate. Both components of the 5-HT response were dose-dependent. Only the excitatory responses were antagonized by metergoline, methysergide and ketanserin. The putative 5-HT agonist, 5-methoxy-N,N-dimethyltryptamine, applied microiontophoretically, depressed the background firing rate and was not antagonized by methysergide. These results demonstrate that 5-HT modifies the responsiveness of vestibular neurons and suggest that at least two mechanisms and maybe two types of receptors are activated by 5-HT in this nucleus.

Serotonin-evoked modifications of the neuronal firing rate in the superior vestibular nucleus: A microiontophoretic study in the rat

Microiontophoretic ejection (10-100 nA) of serotonin (5-hydroxytryptamine) into the superior vestibular nucleus induced modifications of the mean firing rate in 87% of the neurons examined. The responses to 5-hydroxytryptamine application were excitatory in 48% of the cells, inhibitory in 29%, and biphasic (inhibitory/excitatory) in the remaining 10%. The excited neurons were scattered throughout the nucleus; the units inhibited or characterized by biphasic responses were distinctly more numerous in the ventrolateral sector of the nucleus. The magnitude of both excitatory and inhibitory effects was dose-dependent. The excitatory responses to 5-hydroxytryptamine were blocked or greatly reduced by two 5-hydroxytryptamine antagonists, methysergide and ketanserin, or even reversed in many cases. Inhibitory responses were enhanced by simultaneous application of 5-hydroxytryptamine antagonists in half of the units studied. In the remaining units, ketanserin left the response unmodified, whereas methysergide reduced but never quite blocked it. The application of 5-methoxy-N,N- dimethyltryptamine, a 5-hydroxytryptamine agonist more effective on 5-hydroxytryptamine1 than on 5-hydroxytryptamine2 receptors, and of 8-hydroxy-2(di-n-propyl-amino) tetralin, a 5-hydroxytryptamine1A-specific agonist, induced a decrease in the firing rate which was unaffected by methysergide. These results support the hypothesis that 5-hydroxytryptamine exerts various functions throughout the superior vestibular nucleus by various receptors and that the inhibitory action is limited to an area of it.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of noradrenaline on the firing rate of vestibular neurons

The effects of microiontophoretic noradrenaline on the firing rate of neurons located in the vestibular complex have been studied in anaesthetized rats. Eighty-five per cent of the neurons tested in all the vestibular nuclei modified their background firing rate upon noradrenaline application, generally by reducing it (86% of them). In few cases inhibitions were followed by a rebound. Responses were dose-dependent. No significant difference was found between vestibular neurons projecting to the spinal cord and those delivering their fibres to the oculomotor complex. Phentolamine, an alpha-adrenergic antagonist, blocked the noradrenaline-evoked inhibitions, whereas beta-adrenergic antagonist timolol was ineffective or enhanced them. Furthermore, responses were blocked by yohimbine, an alpha 2-adrenergic antagonist, and mimicked by clonidine, an alpha 2-adrenergic agonist, in the majority of neurons. In few cases prazosin, an alpha 1-adrenergic antagonist, was able to antagonize weak inhibitions and phenylephrine, an alpha 1-adrenergic agonist, to evoke an inhibitory effect blocked by prazosin. Isoproterenol, a beta-adrenergic agonist was totally ineffective on the neuronal firing rate. It is concluded that noradrenaline can modify the level of neuronal activity in the vestibular complex by acting mostly, but not exclusively, through alpha 2-adrenergic receptors. An influence of noradrenergic systems on the vestibular function by a direct action of noradrenaline inside the vestibular nuclei is proposed.

Modulation of AMPA receptor‐mediated ion current by pituitary adenylate cyclase‐activating polypeptide (PACAP) in CA1 pyramidal neurons from rat hippocampus

Pituitary adenylate cyclase‐activating polypeptide (PACAP), a neurotrophic and neuromodulatory peptide, was recently shown to enhance NMDA receptor‐mediated currents in the hippocampus (Macdonald, et al. 2005. J Neurosci 25:11374–11384). To check if PACAP might also modulate AMPA receptor function, we tested its effects on AMPA receptor‐mediated synaptic currents on CA1 pyramidal neurons, using the patch clamp technique on hippocampal slices. In the presence of the NMDA antagonist D‐AP5, PACAP (10 nM) reduced the amplitude of excitatory postsynaptic currents (EPSCs) evoked in CA1 pyramidal neurons by stimulation of Schaffer collaterals. Following a paired‐pulse stimulation protocol, the paired‐pulse ratio was unaffected in most neurons, suggesting that the AMPA‐mediated EPSC was modulated by PACAP mainly at a postsynaptic level. PACAP also modulated the currents induced on CA1 pyramidal neurons by applications of either glutamate or AMPA. The effects of PACAP were dose‐dependent: at a 0.5 nM dose, PACAP increased AMPA‐mediated current; such effect was blocked by PACAP 6–38, a selective antagonist of PAC1 receptors. The enhancement of AMPA‐mediated current by PACAP 0.5 nM was abolished when cAMPS‐Rp, a PKA inhibitor, was added to the intracellular solution. At a 10 nM concentration, PACAP reduced AMPA‐mediated current; such effect was not blocked by PACAP 6–38. The inhibitory effect of 10 nM PACAP was mimicked by Bay 55–9837 (a selective agonist of VPAC2 receptors), persisted in the presence of intracellular BAPTA and was abolished by intracellular cAMPS‐Rp. Stimulation‐evoked EPSCs in CA1 neurons were significantly reduced following application of the PAC1 antagonist PACAP 6–38; this result indicates that PAC1 receptors in the CA1 region are tonically activated by endogenous PACAP and enhance CA3‐CA1 synaptic transmission. Our results show that PACAP differentially modulates AMPA receptor‐mediated current in CA1 pyramidal neurons by activation of PAC1 and VPAC2 receptors, both involving the cAMP/PKA pathway; the functional significance will be discussed in light of the multiple effects exerted by PACAP on the CA3‐CA1 synapse at different levels.

Single Muscle Organization of Interposito-rubral Projections*

SummaryIn unanesthetized neuraxis intact cats microstimulation of the interpositus nucleus (IN) which activated a single flexor or extensor muscle in limbs, was used to investigate changes of unitary discharges of rubrospinal (RST) cells. Recordings were made from sites the stimulation of which excited the same muscle activated by the IN (agonist cells), its antagonist (antagonist cells) or heteronymous muscles (heteronymous cells).Cats submitted to chronic cerebellar decortication, acute brachium conjunctivum (BC) section, acute prerubral hemidecerebration or chronic prerubral hemidecerebration and contralateral BC section, were used as controls.It was shown that agonist RST cells were monosynaptically fired from IN, while antagonist cells were inhibited and the heteronymous ones were not influenced.Cerebellar efferents within the BC mediate both excitatory and inhibitory effects, but cerebellar cortex and prerubral structures were not involved in their production.

Neurotransmitter-Mediated Control of Neuronal Firing in the Red Nucleus of the Rat: Reciprocal Modulation between Noradrenaline and GABA

The electrical activity of neurons from the red nucleus, a mesencephalic structure involved in motor control, is under the influence of several neurotransmitters released from afferent fibers and/or from local interneurons. We have investigated the combined effects of gamma-aminobutyric acid (GABA) and noradrenaline (NA), both present at high levels in the red nucleus, on the firing activity of single rubral neurons recorded extracellularly in vivo on anesthetized adult rats. NA inhibited the firing activity of a large part of rubral neurons and induced excitatory or biphasic inhibitory/excitatory effects in a smaller group of cells. Neuronal firing was also inhibited by GABA in all the cells studied. When the effect of GABA was tested during continuous applications of NA, the magnitude of GABA response was modified in 58% of the cells: the effect of GABA was potentiated by NA in half of the responding neurons and was decreased in the remaining half. NA-induced potentiation of GABA response was mimicked by the alpha(2)-adrenoceptor agonist clonidine and was abolished by the alpha(2)-adrenoceptor antagonist yohimbine. On the other side, the decrease of GABA response was reproduced by the beta-adrenoceptor agonist isoprenaline and was blocked by timolol, an antagonist of beta-adrenoceptors. Neuronal firing activity was reduced by nipecotic acid, an inhibitor of GABA reuptake mechanism, and was instead increased during application of the GABA(A) receptor antagonist bicuculline, suggesting that rubral neurons in vivo were under tonic control by endogenous GABA. Both the inhibitory and the excitatory effects of NA were reduced in the presence of nipecotic acid and were instead potentiated during application of bicuculline, suggesting that NA responses were modified by endogenous GABA. Taken together, our results indicate a reciprocal modulation between the effects of GABA and NA on neuronal firing activity in the red nucleus of the rat: GABA depresses the responsiveness of rubral neurons to NA, whereas NA is able either to potentiate or to decrease the effects of GABA by activation of alpha(2)- and beta-adrenoceptors, respectively. The functional significance of such interaction, as well as the possible implication in diseases affecting motor control, will be discussed.

Effects of motor cortex and single muscle stimulation on neurons of the lateral vestibular nucleus in the rat

The neuronal responses to stimulation of motor cortical sites and of forelimb single muscles were studied in the lateral vestibular nucleus of anaesthetized rats. Of the 228 neurons tested for response to stimulation of contralateral motor cortex, 63% responded to cortical sites controlling extensor muscles and 30% to those controlling flexors. The corresponding figures for responders to ipsilateral stimulation were 34 and 21%. Vestibulospinal units responded to cortical sites controlling extensor and flexor muscles whereas the remaining lateral vestibular nucleus neurons, very reactive to cortical sites controlling extensor muscles, responded little to contralateral and not at all to ipsilateral cortical sites controlling flexor muscles. The effects evoked by contralateral cortical sites controlling extensors varied, those induced by cortical sites controlling flexors were inhibitory in 77% of cases. The responses to ipsilateral motor cortex stimulation differed not so much by cortical sites controlling extensor or flexor muscles as by whether the neuron was in the dorsal or ventral zone of the lateral vestibular nucleus: mixed in the former, all inhibitory in the latter. Of the lateral vestibular nucleus units tested for response to stimulation of ipsilateral or contralateral forelimb distal muscles, only 11% responded. All the vestibulospinal units responsive to muscle stimulation lay in the dorsal zone of the nucleus. The remainder, dorsal or ventral, were not responsive to contralateral muscles. Single lateral vestibular nucleus cells influenced both by ipsilateral muscle and by contralateral motor cortex made up 24% of the pool, vestibulospinal and non-vestibulospinal. They fell into three groups: responsive to one or both structures but responding more strongly to combined stimulation; responsive to each of the two structures but showing a response to combined stimulation not significantly different from that evoked by the cortex alone; responsive only to combined stimulation. The lateral vestibular nucleus units included in these three groups accounted for 29% of those tested for response to extensor muscles and cortical sites controlling extensors and 15% of those tested for response to flexor muscles and cortical sites controlling flexors. Twenty-five per cent of the vestibulospinal neurons responded both to contralateral muscles and to ipsilateral motor cortex stimulation but none of the non-vestibulospinal neurons responded to both. All the responders to both were in the dorsal zone of the lateral vestibular nucleus and responded to extensor stimuli, always in the same way. These results indicate that motor cortex output exerts a major influence on lateral vestibular nucleus discharges, while the muscle afferents have a modulatory influence on the lateral vestibular nucleus responses to cortex.(ABSTRACT TRUNCATED AT 400 WORDS)

Alpha2- and beta-adrenoceptors differentially modulate GABAA- and GABAB-mediated inhibition of red nucleus neuronal firing

In mesencephalic red nucleus (RN), GABA-induced inhibition of neuronal firing is modulated by noradrenaline acting on alpha2- and beta-adrenoceptors. Since both GABAA and GABAB receptors are present in the rat RN, we have recorded the firing activity of RN neurons in vivo from anaesthetized rats to study how GABAA- and GABAB-mediated effects are modulated by either alpha2- or beta-adrenoceptor activation. Both the GABAA agonist isoguvacine and the GABAB agonist baclofen depressed the firing of RN neurons. During simultaneous application of clonidine, an alpha2-adrenoceptor agonist, half of the isoguvacine- and baclofen-mediated responses were modified: isoguvacine-mediated inhibition was enhanced by 97% without any change in effect duration, whereas baclofen responses were either increased or slightly reduced in the same number of cases. Application of isoprenaline, a beta-adrenoceptor agonist, increased isoguvacine effect in 66% of neurons without modifying effect duration; the amount of increase (43%) was significantly lower than that induced by clonidine. On the other hand, in the presence of isoprenaline, baclofen response was reduced in 72% of neurons with respect to both the amount (52%) and the duration (34%) of effect. Taken together, these results indicate that alpha2-adrenoceptors mainly enhance GABAA-induced inhibition and induce mixed effects on GABAB response; on the other side, beta-adrenoceptors exert an opposite modulation on GABA effects, respectively, enhancing and depressing GABAA- and GABAB-mediated responses.

Low concentrations of gaba reduce accommodation in primary afferent neurons by an action at GABAB receptors

The pattern of accommodation of spike activity during sustained membrane depolarization was investigated in primary afferent neurons recorded intracellularly in vitro in the rat. We show that gamma-aminobutyric acid (GABA) and baclofen reduce accommodation in some fast conducting dorsal root ganglion neurons. This effect was restricted to those A delta cells with axons displaying a rather fast conduction velocity (15-25 m/s). GABA-induced blockade of accommodation was not observed in large A beta neurons. Pharmacological studies with baclofen, as opposed to isoguvacine, indicate that this effect is due to GABAB receptors activation. The effect is also shown to be resistant to bicuculline antagonism. In slow conducting afferents, GABAB receptor activation is known to shorten the CA2+ component of action potentials. By contrast, no such component was observed in the A delta cells studied. Furthermore, Ca2+-activated K+ conductances are not implicated in the reduction of accommodation caused by GABAB receptor activation. In conjunction with the actual knowledge about the distribution of GABA receptors on primary afferents, our result indicates that GABAA and GABAB receptors coexist on all categories of A delta and C primary afferents in the rat.

Motor responses evoked by microstimulation of cerebellar interpositus nucleus in cats submitted to dorsal rhizotomy

In cats with C4-T2 unilateral dorsal rhizotomy, stimulation of interpositus nucleus of the cerebellum, ipsilateral to the deafferented side, elicits in forelimbs single muscle contractions, which display threshold, latencies and spatial-temporal characteristics similar to those of the muscular responses produced in the other forelimb, upon stimulation of interpositus nucleus of the intact side. Motor effects induced by the interpositus nucleus-stimulation on the deafferented side disappear following red nucleus lesions. Single muscle contractions triggered from interpositus nucleus are, therefore, mediated by impulses impinging on alpha-motoneurons, via the rubrospinal pathway.