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The actions of monoamines and distribution of noradrenergic and serotoninergic contacts on different subpopulations of commissural interneurons in the cat spinal cord.

Modulatory actions of monoamines were investigated on spinal commissural interneurons which coordinate left-right hindlimb muscle activity through direct projections to the contralateral motor nuclei. Commissural interneurons located in Rexed lamina VIII, with identified projections to the contralateral gastrocnemius-soleus motor nuclei, were investigated in deeply anaesthetized cats. Most interneurons had dominant input from either the reticular formation or from group II muscle afferents; a small proportion of neurons had input from both. Actions of ionophoretically applied serotonin and noradrenaline were examined on extracellularly recorded spikes evoked monosynaptically by group II muscle afferents or reticulospinal tract fibres. Activation by reticulospinal fibres was facilitated by both serotonin and noradrenaline. Activation by group II afferents was also facilitated by serotonin but was strongly depressed by noradrenaline. To investigate the possible morphological substrates of this differential modulation, seven representative commissural interneurons were labelled intracellularly with tetramethylrhodamine-dextran and neurobiotin. Contacts from noradrenergic and serotoninergic fibres were revealed by immunohistochemistry and analysed with confocal microscopy. There were no major differences in the numbers and distributions of contacts among the interneurons studied. The findings suggest that differences in modulatory actions of monoamines, and subsequent changes in the recruitment of subpopulations of commissural interneurons in various behavioural situations, depend on intrinsic interneuron properties rather than on the patterns of innervation by monoaminergic fibres. The different actions of noradrenaline on different populations of interneurons might permit reconfiguration of the actions of the commissural neurons according to behavioural context.

Action Potentials↗

Influence of the hippocampus on interneurons of the rat prefrontal cortex.

The hippocampus and prefrontal cortex (PFC), two structures implicated in learning and memory processes, are linked by a direct hippocampo-prefrontal pathway. It has been shown that PFC pyramidal cells receive monosynaptic excitatory inputs from the hippocampus and, in this study, we sought to determine the influence of the hippocampus on PFC interneurons in anesthetized rats. Extracellular recordings were coupled to juxtacellular injections of neurobiotin or biotinylated dextran amine to morphologically differentiate interneurons from pyramidal cells. In all cases, the action potentials of labeled interneurons were of shorter duration (< 0.70 ms) than those of identified pyramidal cells (> 0.70 ms). Single pulse stimulation of the hippocampal CA1/subiculum region induced an excitatory response in 70% of recorded interneurons in the prelimbic and medial-orbital areas of the PFC. In contrast to the one to two action potentials generated by pyramidal cells, an important group of interneurons fired a burst of action potentials in response to hippocampal stimulation. A large proportion of these excitatory responses was probably monosynaptic as their latency is consistent with the conduction time of the hippocampo-prefrontal pathway. In addition, when both a pyramidal cell and an interneuron were simultaneously recorded and both responded to stimulation, the interneuron consistently fired before the pyramidal cell. In conclusion, the hippocampus exerts a direct excitatory influence on PFC interneurons and is thus capable of feedforward inhibition of pyramidal cells. Hippocampal output is spatially and temporally focalized via this inhibitory process and consequently could facilitate the synchronization of a specific subset of PFC neurons with hippocampal activity.

Action Potentials↗

Cell-type specific GABA synaptic transmission and activity-dependent plasticity in rat hippocampal stratum radiatum interneurons.

Abstract In hippocampal pyramidal cells, the efficacy of synaptic transmission at gamma-aminobutyric acid (GABA)ergic synapses, is modulated by activity. However, whether such plasticity occurs at inhibitory synapses on interneurons remains largely unknown. Using whole-cell voltage-clamp recordings of inhibitory postsynaptic currents (IPSCs) in Sprague-Dawley rat hippocampal slices, we examined whether GABA synapses of stratum radiatum interneurons were affected by stimulation protocols known to alter efficacy at inhibitory synapses of CA1 pyramidal cells. Monosynaptically evoked IPSCs (eIPSCs) exhibited different properties with significantly faster kinetics, higher coefficients of variation, a current-voltage (I-V) relationship shifted to depolarized values and a smaller paired-pulse depression, in interneurons than in pyramidal cells. GABA synapses on interneurons also showed a different capacity for plasticity. Indeed, theta-burst stimulation induced a long-term potentiation of eIPSCs in both cell types, but the induction mechanisms differed in interneurons, as it was not affected by antagonists of GABAB receptors and group I/II metabotropic glutamate receptors (mGluRs). Furthermore, 100-Hz tetanization selectively elicited a short-term depression of eIPSCs in pyramidal cells. A postsynaptic depolarization produced a transient suppression of eIPSCs (depolarization-induced suppression of inhibition) in pyramidal cells but not in interneurons. Spontaneous IPSCs were similarly reduced following depolarization in pyramidal cells, but not in interneurons. These results indicate that GABA synapses of stratum radiatum interneurons exhibit different properties and capacity for activity-dependent plasticity than those of pyramidal cells. This cell-type specific mode of transmission and adaptive regulation of GABA synapses may contribute to hippocampal plasticity and functions.

Animals↗

Interneurones in pathways from group II muscle afferents in the lower-lumbar segments of the feline spinal cord.

Interneurones receiving excitatory input from group II muscle afferents of hindlimb nerves and located in the lower-lumbar (L6-L7) segments of the cat spinal cord were investigated using both extracellular and intracellular recording. The interneurones were located mainly in the lateral parts of laminae IV-VII, dorsal and lateral to the main region in which interneurones with input from group I muscle afferents are located. Almost half the sample of interneurones (38 of 76) were characterized by an ipsilateral ascending projection within the lateral funiculus to the L4 level. The most powerful group II excitation was produced by afferents of the quadriceps and deep peroneal muscle nerves (which discharged 70-80% of extracellularly recorded neurones) while group II afferents of tibialis posterior, posterior biceps-semitendinosus and gastrocnemius soleus were also highly effective (discharging 45-55% of extracellularly recorded neurones). A proportion of intracellularly recorded group II EPSPs were monosynaptic. Seventy-five per cent of the extracellularly recorded interneurones were discharged by group II afferents of two or more muscle nerves and 43% by afferents of three or more nerves. Group I muscle afferents evoked small EPSPs in over one-quarter of the intracellularly recorded interneurones and virtually all were strongly excited by cutaneous afferents. Evidence of excitatory input from joint, interosseous and group III muscle afferents was also observed. The properties of the interneurones are compared with those of others in the lumbosacral segments and the possibility that they may function as last-order premotor interneurones is discussed.

Animals↗

Concomitant changes in formaldehyde-induced fluorescence of dopamine interneurones and in slow inhibitory post-synaptic potentials of the rabbit superior cervical ganglion, induced by stimulation of the preganglionic nerve or by a muscarinic agent.

1. Dopamine was identified by formaldehyde histochemistry and cytospectrofluorometry in the rabbit's superior cervical ganglion. Dopamine was localized to the intraganglionic ;small intensely fluorescent' cells, and also to the characteristically beaded fibres forming a network in close contact with virtually all ganglion cell bodies. The extensive beaded fibres are therefore presumed to be processes of the small intensely fluorescent cells.2. Changes in the dopamine content of these interneurones were studied by recording alterations in their relative fluorescence intensity in conjunction with changes in the slow inhibitory post-synaptic potential (s.-i.p.s.p.) response of the ganglion to orthodromic nerve input.3. Dopamine content was lower after several hours in vitro even without special stimulation; this was in accord with a regularly observed spontaneous reduction of the s.-i.p.s.p. response.4. After a period of conditioning stimulation of the preganglionic nerve, in the presence of an anticholinesterase agent (eserine) and an inhibitor of catecholamine synthesis (alpha-methyl-p-tyrosine), the s.-i.p.s.p. was selectively and markedly reduced. The dopamine fluorescence in the small intensely fluorescent cell interneurones was also significantly reduced, to a mean value of about 55 or 60% of the fluorescence in the dopamine interneurones of the paired but unstimulated control ganglion. A significant reduction in dopamine fluorescence was always accompanied by a marked loss of s.-i.p.s.p. response; the reverse was not always true.5. Treatment with the muscarinic agent bethanechol for 30 min, with no alpha-methyl-p-tyrosine or eserine present, similarly resulted in reductions in the s.-i.p.s.p. response of the ganglia and in the formaldehyde-induced fluorescence of the dopamine interneurones.6. A functional uptake of extrinsic dopamine by the dopamine interneurones was also demonstrated: temporary exposure to dopamine restored a large fraction of both the s.-i.p.s.p. response and the dopamine fluorescence of the small intensely fluorescent cells, in ganglia already subjected either to the conditioning stimulation of the preganglionic nerve or to the action of bethanechol.7. It is concluded that (a) preganglionic impulses, by a cholinergic muscarinic synaptic action, can induce a release of dopamine from dopamine interneurones (small intensely fluorescent cells) in the superior cervical ganglion, (b) the ability of the ganglion to respond with a s.-i.p.s.p. to orthodromic input may be viewed as being dependent on the supply of functionally releasable dopamine in these interneurones, (c) the functionally releasable transmitter in vitro appears to comprise roughly 50% of the total dopamine content of the interneurones, and (d) the results fulfil some of the requirements of the hypothesis that a dopamine interneurone is activated muscarinically by preganglionic nerve impulses and mediates the production of s.-i.p.s.p. in sympathetic ganglion cells.

Action Potentials↗

The same interneurones mediate inhibition of dorsal spinocerebellar tract cells and lumbar motoneurones in the cat.

The aim of the study was to investigate whether inhibition of dorsal spinocerebellar tract (d.s.c.t.) cells evoked from group I afferents is mediated by the same interneurones which mediate the non-reciprocal inhibition of hind-limb motoneurones. The origin of inhibition of d.s.c.t. cells from group I afferents was compared in intact preparations, after lesions of the dorsal funiculi (when it could only be mediated by lower lumbar interneurones) and after lesions of the lateral and ventral funiculi (when it would be expected to be evoked by upper lumbar interneurones). In all three preparations extensors were the most common source of inhibition, as in motoneurones. Lower lumbar interneurones inhibiting d.s.c.t. cells were found to be co-excited by group I (Ia and/or Ib) and cutaneous and joint afferents, and by rubrospinal tract fibres, as are interneurones mediating inhibition of motoneurones. Co-excitation by group I and rubrospinal fibres was also found for upper lumbar interneurones. I.p.s.p.s were evoked in hind-limb motoneurones from within Clarke's column in cats with the dorsal funiculi cut between L4 and L5 segments; they were evoked at thresholds as low as 2 microA, i.e. by stimuli with very local actions. The latencies of these i.p.s.p.s were short enough to allow them to be evoked monosynaptically via axonal branches of the same interneurones which projected to Clarke's column. Correspondingly, i.p.s.p.s were evoked in d.s.c.t. cells from within motor nuclei in L7 segments; they were evoked at similarly low thresholds and with similar latencies. In confirmation of previous reports i.p.s.p.s of Ia origin evoked in d.s.c.t. cells were not found to be depressed by Renshaw cells, which excludes their mediation by interneurones responsible for Ia reciprocal inhibition. The study leads to the conclusion that the inhibition of d.s.c.t. cells from group I afferents is, at least in part, collateral to the non-reciprocal inhibition of lumbar motoneurones.

Animals↗

Post-synaptic potentials in a population of motoneurones following activity of single interneurones in the cat.

The technique of recording post-synaptic potentials from a population of motoneurones, by recording from ventral roots perfused with isotonic sucrose, has been applied to investigate the action of single last-order interneurones; the target motoneurones were in either caudal L7 or S1 segments. Using spike-triggered averaging, the inhibitory action of 70% of previously identified last-order interneurones (Renshaw cells and lamina VII Ia inhibitory interneurones) has been detected. Previous observations had suggested that interneurones mediating disynaptic non-reciprocal inhibition from group I muscle afferents should be characterized by (i) location in laminae V-VI, (ii) monosynaptic group I input and (iii) ascending collateral axonal projection to upper lumbar segments. 65% of interneurones with these characteristics were found to inhibit motoneurones. In addition, spike-triggered averaging from this group of laminae V-VI interneurones sometimes revealed a depolarizing potential which preceded the inhibitory potential evoked by the interneurone. The depolarizing potential is interpreted as being due to the action of some presynaptic fibres which branch to innervate both the investigated interneurones and motoneurones.

Animals↗

Reciprocal inhibitory interneurones in the Xenopus embryo spinal cord.

The mechanism of reciprocal inhibition between antagonistic motor centres during swimming in the paralysed Xenopus embryo has been investigated further. Paired intracellular recordings have been made from interneurones and motoneurones in an attempt to identify neurones which make direct inhibitory synapses onto motoneurones on the opposite side of the spinal cord. A physiological class of inhibitory interneurones is described which, when stimulated by intracellular current passage, evoke short-latency, probably monosynaptic, strychnine-sensitive inhibitory potentials in contralateral motoneurones. These inhibitory interneurones fire once per swimming cycle in phase with the ipsilateral motor root discharge. They therefore have a pattern of activity which would cause them to inhibit motoneurones of the antagonistic motor centre at an appropriate part of the swimming cycle. The intracellular injection of horseradish peroxidase (HRP) has allowed the morphology of these inhibitory interneurones to be characterized. They have unipolar cell bodies with a thick proximal process with short dendrites which crosses the spinal cord ventrally and then bifurcates with one axonal branch ascending into the hind brain and the other descending the spinal cord. These anatomical features are typical of the 'commissural interneurones' first described by Roberts & Clarke (1982). There are also some inhibitory interneurones which can inhibit motoneurones on the same side of the spinal cord. At least some of these interneurones may be commissural interneurones with ipsilateral axons and they may play a role in the generation of the swimming rhythm.

Animals↗

Lamina VIII interneurones interposed in crossed reflex pathways in the cat.

The location of a group of interneurones projecting to contralateral motor nuclei has been established using retrograde transneuronal transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP). After labelling the motoneurones of semitendinosus, medial gastrocnemius or quadriceps muscles, interneurones which were secondarily labelled were found in lamina VIII and in the neighbouring narrow strip of lamina VII. They were found to be distributed from the 4th lumbar to the 1st sacral segments, with the highest concentration in the 6th and 7th lumbar segments and at the border between the 4th and 5th lumbar segments. The electrophysiological properties of lamina VIII interneurones of the 6th lumbar segment have been investigated using both extracellular and intracellular recording. Many of these interneurones could be antidromically activated following weak stimuli applied in contralateral motor nuclei. Post-synaptic potentials were evoked from a variety of primary afferents including group I muscle afferents. However, when present, the post-synaptic potentials (p.s.p.s) of group I origin were of considerably smaller amplitudes than p.s.p.s. evoked from higher threshold muscle or cutaneous afferents and smaller than p.s.p.s. which followed stimulation of the spinal cord at the thoracic level. P.s.p.s. from the latter two sources appear to constitute the main input to lamina VIII interneurones. Group I input has been found in forty lamina VIII interneurones. These were usually affected by either ipsilateral or contralateral group I afferents and only exceptionally by both. Excitatory post-synaptic potentials (e.p.s.p.s) from ipsilateral afferents were evoked in about twice as many neurones as e.p.s.p.s from the contralateral afferents. E.p.s.p.s were often accompanied by inhibitory post-synaptic potentials (i.p.s.p.s). Group Ia afferents appeared to contribute to both e.p.s.p.s and i.p.s.p.s, whether these were evoked from ipsilateral or from contralateral afferents. In several cases Ia afferents were as effective as all group I afferents while in other cases Ib afferents appeared to be an important or even the exclusive source of the p.s.p.s. The latencies of e.p.s.p.s indicated that they were evoked mono-, di- or trisynaptically from ipsilateral group I afferents and di- or trisynaptically from contralateral afferents, I.p.s.p.s appeared to be evoked via pathways with only one additional interneurone. About one-third of all the intracellularly investigated lamina VIII interneurones were both affected by group I afferents and antidromically activated from the contralateral motor nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Post-synaptic actions of midlumbar interneurones on motoneurones of hind-limb muscles in the cat.

1. The hypothesis that interneurones in the 4th lumbar segment (L4) are interposed between group I and group II afferents and hind-limb motoneurones has been tested. Action potentials of single interneurones were induced by ionophoretically applied homocysteate and recorded in parallel with post-synaptic potentials in motoneurones; the latter were recorded from motor axons in the ventral root of the first sacral segment as population potentials, using the sucrose gap technique. 2. The action potentials of twenty-four L4 interneurones were found to be followed by either e.p.s.p.s. or i.p.s.p.s in motoneurones. The latencies of the majority of these p.s.p.s were consistent with monosynaptically evoked excitation or inhibition of motoneurones since they exceeded the latencies of antidromic activation of the interneurones from the S1 motor nuclei by only a fraction of a millisecond. 3. The dominant input to both the excitatory and the inhibitory interneurones was from group II muscle afferents, in particular from the quadriceps nerve. The latencies of excitation of the interneurones by these afferents indicated a monosynaptic coupling between them. The same interneurones were co-excited by group I and cutaneous afferents and by descending fibres. 4. We conclude that not only excitation but also inhibition of hind-limb motoneurones from group II afferents may be mediated disynaptically and that interneurones in the 4th lumbar segment contribute to both.

Action Potentials↗

The monosynaptic excitatory connections of single trigeminal interneurones to the V motor nucleus of the rat.

1. We have used the extracellular spike-triggered averaging method to identify a population of trigeminal interneurones that make monosynaptic connections within the V motor nucleus. The experiments were performed on rats anaesthetized with pentobarbitone (60 mg/kg I.V.; supplementary doses given as necessary to maintain a deep level of anaesthesia). 2. A tungsten microelectrode (tip exposure of some 200 microns) was inserted into the masseter motoneurone pool to allow recording of extracellular activity. A glass electrode filled with DL-homocysteic acid was used to make simultaneous extracellular recordings of the firing of single neurones in the region immediately caudal to the motor nucleus. 3. Fifty-eight out of 166 interneurones tested gave unitary extracellular fields in the motor nucleus. The responses consisted of a terminal spike (presynaptic spike) followed by a negative field of duration approximately 3 ms and amplitude 0.4-10.8 microV. The mean latency between the positive peak of the terminal spike and the onset of the field (synaptic delay) was 0.43 ms (S.D. = 0.10 ms), and the mean latency from the onset of the interneurone spike to the positive peak of the presynaptic spike was 0.35 ms (S.D. = 0.22 ms). Thus the interneurones project directly to the motor nucleus where they then make monosynaptic connections. 4. The negative extracellular fields were taken to reflect an excitatory synaptic input onto neurones within the motor nucleus. The fields were of maximum amplitude at the point of maximum masseter motoneurone antidromic field, suggesting that the connection may be onto elevator motoneurones. 5. The location of the interneurone appeared to the main factor governing the likelihood of obtaining a field. Interneurones located more than 400 microns from the caudal border of the motor nucleus rarely produced fields whereas most of those located within this distance gave fields. This pattern of distribution is strikingly similar to that of trigeminal interneurones labelled by the transneuronal transport of wheatgerm agglutinin-horseradish peroxidase after an intramuscular injection of the tracer into the masseter muscle. We suggest that this provides electrophysiological support for the suggestion that the tracer does indeed label interneurones by means of retrograde transsynaptic transport.

Animals↗

Control of frequency during swimming in Xenopus embryos: a study on interneuronal recruitment in a spinal rhythm generator.

1. In Xenopus embryos, the frequency of natural and fictive swimming usually drops slowly as swimming continues but can increase following stimulation of the skin or dimming of the illumination. We have investigated whether such increases are associated with an increase in the number of neurones active at higher frequencies. 2. Recordings from ventral presumed motoneurones show that these were reliably active at all swimming frequencies. 3. Recordings from more dorsal presumed interneurones showed that in the majority of these firing probability decreased as a function of swimming frequency. Dye-filled microelectrodes were used to show that some of these neurones had the anatomy of known classes of excitatory and inhibitory premotor interneurones. 4. If skin stimulation is given at appropriate phases of the swimming cycle, it can lead to a transient increase in frequency. Recordings from silent premotor interneurones during such stimulation show that they can be recruited to fire during the post-stimulus frequency increases. 5. It was possible that spike failure in the interneurones could have been due to damage by the recording microelectrodes. We therefore measured the amplitudes and probability of occurrence of rhythmic 'on-cycle' IPSPs which occur in sensory interneurones and 'on-cycle' IPSPs which sometimes occur in motoneurones during fictive swimming. Both decreased in amplitude and could fail as frequency dropped, providing further evidence that the number of inhibitory interneurones firing on each cycle of swimming is a function of frequency. 6. We conclude that premotor rhythm-generating interneurones are not active on all cycles of swimming and that their probability of firing action potentials increases with swimming frequency. This suggests that swimming frequency is determined in part by the number of premotor interneurones which are active.

Animals↗

Candidate interneurones mediating group I disynaptic EPSPs in extensor motoneurones during fictive locomotion in the cat.

In the present study we sought to find interneurones responsible for the group I-evoked disynaptic excitation of hindlimb extensor motoneurones that occurs during fictive locomotion. Locomotion was produced by stimulation of the mesencephalic locomotor region (MLR) in decerebrate paralysed cats in which activation of ankle extensor group I afferents evoked a disynaptic excitation of motoneurones during the extension phase of fictive locomotion. Extracellular recordings were used to locate interneurones fulfilling all, or five of the six following criteria: (i) weak or no response to stimulation of extensor group I afferents in the absence of locomotion; (ii) strong group I activation during locomotion; (iii) group I activation at monosynaptic latencies; (iv) strong group I activation during only the extensor phase of locomotion; and (v) antidromic activation from the extensor motor nuclei; but (vi) no antidromic activation from rostral spinal segments. Candidate excitatory interneurones were located in mid to caudal parts of the L7 segments in areas where monosynaptic field potentials were evoked by group I afferents, within 2 mm of the stimulation site in the ventral horn from which they were antidromically activated. All were activated during extension by stimulation of group I afferents in extensor nerves. In the absence of peripheral nerve stimulation, six of the seven candidate excitatory interneurones were rhythmically active with maximal activation during the extension phase of fictive locomotion. Rhythmic activity during extension was also seen in five additional interneurones located near candidate interneurones but not activated by group I strength stimulation of the tested nerves. We suggest that the lumbosacral interneurones located in the intermediate laminae that can be activated by extensor group I afferents during the extension phase are a previously unknown population of interneurones, and may mediate group I-evoked disynaptic excitation of extensor motoneurones. Their rhythmic activity suggests that they also provide central excitatory drive to extensor motoneurones during locomotion.

Afferent Pathways↗

Molecular diversity of neocortical GABAergic interneurones.

In addition to being the main source of inhibition in the adult brain, GABAergic interneurones are instrumental in pacing the activity of large ensembles of principal cells. GABAergic interneurones have unique features that enable them to contribute to the generation of synchronized network activity thereby shaping principal cell behaviour. Whereas the anatomical and physiological characteristics of certain interneuronal types have been studied extensively over the last decades, the molecular diversity of interneurones is a more recent focus of investigation in this field. Molecular cloning and expression analysis of many receptor families often revealed differential expression in GABAergic interneurones and pyramidal cells. Here we review recent findings regarding the molecular diversity of GABAergic interneurones in the neocortex. Better knowledge about differential gene expression in GABAergic interneurones is the basis for further investigations aimed at understanding the contribution of specific proteins in interneurones to network function.

Animals↗

Functional differentiation and organization of feline midlumbar commissural interneurones.

Interneurones interconnecting the two sides of the spinal cord (commissural interneurones) are critically important for interlimb coordination, but little is known about their organization. We have examined the inputs to commissural interneurones located in the midlumbar segments with projections to contralateral motor nuclei, aiming to determine whether they form distinct subpopulations. Based on intracellular records from 78 interneurones, two major non-overlapping subpopulations were identified: one monosynaptically excited by group II muscle afferents (n=10), the other monosynaptically excited by reticulospinal neurones (n=52). Monosynaptic input from group I muscle afferents and/or from vestibulospinal tract neurones was found in those with monosynaptic reticulospinal, but not group II input, and in a few other neurones (n=6). Only disynaptic input from these sources was found in the remaining 10 interneurones. Disynaptic excitatory input from ipsilateral and contralateral muscle afferents and from descending tracts was distributed less selectively and might mediate coexcitation of interneurones with monosynaptic afferent or descending input. The dominant disynaptic and polysynaptic input was, however, inhibitory. IPSPs were evoked from the descending tracts in a high proportion of the commissural interneurones that were monosynaptically excited by group II afferents (55%) and from group II afferents in a high proportion of the commissural interneurones that were monosynaptically excited by reticulospinal fibres (78%). This distribution suggests that the two subpopulations are activated differentially, rather than being coactivated, in either centrally initiated movements or reflex adjustments. This would be consistent with the previous demonstration that noradrenaline differentially affects commissural neurones of the two subpopulations.

Animals↗

5-HT Modulation of identified segmental premotor interneurons in the lamprey spinal cord.

Ipsilaterally projecting spinal excitatory interneurons (EINs) generate the hemisegmental rhythmic locomotor activity in lamprey, while the commissural interneurons ensure proper left-right alternation. 5-HT is a potent modulator of the locomotor rhythm and is endogenously released from the spinal cord during fictive locomotion. The effect of 5-HT was investigated for three segmental premotor interneuron types: EINs, commissural excitatory and commissural inhibitory interneurons. All three types of interneurons produced chemical postsynaptic potentials in motoneurons, but only those from EINs had an electrical component. The effect of 5-HT was studied on the slow afterhyperpolarization, involved in spike frequency regulation, and on the segmental synaptic transmission to motoneurons. 5-HT induced a reduction in the slow afterhyperpolarization and a depression of synaptic transmission in all three types of segmental interneurons. Thus 5-HT is a very potent modulator of membrane properties and synaptic transmission of last-order segmental premotor interneurons. Such modulation of locomotor network interneurons can partially account for the observed effects of 5-HT on the swimming pattern in lamprey.

Action Potentials↗

Myomodulin increases Ih and inhibits the NA/K pump to modulate bursting in leech heart interneurons.

In the medicinal leech, a rhythmically active 14-interneuron network composes the central pattern generator for heartbeat. In two segmental ganglia, bilateral pairs of reciprocally inhibitory heart interneurons (oscillator interneurons) produce a rhythm of alternating bursts of action potentials that paces activity in the pattern-generating network. The neuropeptide myomodulin decreases the period of this bursting and increases the intraburst spike frequency when applied to isolated ganglia containing these oscillator interneurons. Myomodulin also decreases period, increases spike frequency, and increases the robustness of endogenous bursting in synaptically isolated (with bicuculline) oscillator interneurons. In voltage-clamp experiments using hyperpolarizing ramps, we identify an increase in membrane conductance elicited by myomodulin with the properties of a hyperpolarization-activated current. Voltage steps confirm that myomodulin indeed increases the maximum conductance of the hyperpolarization-activated current I(h). In similar experiments using Cs(+) to block I(h), we demonstrate that myomodulin also causes a steady offset in the ramp current that is not associated with an increase in conductance. This current offset is blocked by ouabain, indicating that myomodulin inhibits the Na/K pump. In current-clamp experiments, when I(h) is blocked with Cs(+), myomodulin decreases period and increases spike frequency of alternating bursting in synaptically connected oscillator interneurons, suggesting that inhibiting the Na/K pump modulates these burst characteristics. These observations indicate that myomodulin decreases period and increases spike frequency of endogenous bursting in synaptically isolated oscillator heart interneurons and alternating bursting of reciprocally inhibitory pairs of interneurons, at least in part, by increasing I(h) and by decreasing the Na/K pump.

Action Potentials↗

Organization of receptive fields of spiking local interneurons in the locust with inputs from hair afferents.

The receptive fields of spiking local interneurons in the locust were defined by making intracellular recordings from them while stimulating mechanoreceptors on the surface of a hindleg. All the interneurons tested have their cell bodies near the ventral midline, in the so-called "midline" group. Those described here receive inputs only from external mechanoreceptors; others receive inputs from internal proprioceptors alone or from receptors of both kinds. The receptors on the surface of a hindleg that contribute to the receptive field of an interneuron may be clustered together in a discrete area or be distributed in separate regions that provide either excitation or inhibition. An interneuron may have a receptive field that is wholly excitatory or one with both excitatory and inhibitory regions. Excitatory but not inhibitory effects are mediated by direct connections between afferents and interneurons. The longitudinal boundaries of most receptive fields occur along one of the major axes of the leg. For example, hairs on the anterior half of a leg can be excitatory, while hairs on the posterior half are inhibitory; or those on the dorsal half are excitatory, and those on the ventral half are inhibitory. The proximal-distal boundaries of a receptive field often correspond to the articulations between the segments of a leg, although they may also occur within a segment of a leg where there are no other obvious anatomical discontinuities. The receptive fields of these interneurons often overlap, and an individual afferent from a hair excites more than one interneuron. In this way a particular region on the surface of a hindleg may be mapped onto as many as 12 interneurons. The size of a receptive field is not correlated with its position along the proximal-distal axis of the leg, but the smallest fields occur on the distal tibia or span the femorotibial joint.

Animals↗