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To assess when vestibulosympathetic projections become functional postnatally, and to establish a preparation in which vestibulosympathetic circuitry can be characterized more precisely, we used an optical approach to record VIIIth nerve-evoked synaptic inputs to thoracic sympathetic preganglionic neurons (SPNs) in newborn mice. Stimulation of the VIIIth nerve was performed in an isolated brainstem-spinal cord preparation after retrogradely labelling with the fluorescent calcium indicator Calcium Green 1-conjugated dextran amine, the SPNs and the somatic motoneurons (MNs) in the thoracic (T) segments T2, 4, 6, 8, 10 and 12. Synaptically mediated calcium responses could be visualized and recorded in individual SPNs and MNs, and analysed with respect to latency, temporal pattern, magnitude and synaptic pharmacology. VIIIth nerve stimulation evoked responses in all SPNs and MNs investigated. The SPN responses had onset latencies from 90 to 200 ms, compared with much shorter latencies in MNs, and were completely abolished by mephenesin, a drug that preferentially reduces polysynaptic over monosynaptic transmission. Bicuculline and picrotoxin, but not strychnine, increased the magnitudes of the SPN responses without changing the onset latencies, suggesting a convergence of concomitant excitatory and inhibitory synaptic inputs. Lesions strategically placed to test the involvement of direct vestibulospinal pathways versus indirect pathways within the brainstem showed that vestibulosympathetic inputs in the neonate are mediated predominantly, if not exclusively, by the latter. Thus, already at birth, synaptic connections in the vestibulosympathetic reflex are functional and require the involvement of the ventrolateral medulla as in adult mammals.

Proper control of movement and posture occurs partly via descending projections from the vestibular nuclei to spinal motor circuits. Days before birth in rodents, vestibulospinal neurons develop axonal projections that extend to the spinal cord. How functional these projections are just after birth is unknown. Our goal was to assess the overall functional organization of vestibulospinal inputs to spinal motoneurons in a brainstem–spinal cord preparation of the neonatal mouse (postnatal day (P) 0–5). Using calcium imaging, we recorded responses evoked by electrical stimulation of the VIIIth nerve, in many motoneurons simultaneously throughout the spinal cord (C2, C6, T7, L2 and L5 segments), in the medial and lateral motor columns. Selective lesions in the brainstem and/or spinal cord distinguished which tracts contributed to the responses: those in the cervical cord originated primarily from the medial vestibulospinal tracts but with a substantial contribution from the lateral vestibulospinal tract; those in the thoracolumbar cord originated exclusively from the lateral vestibulospinal tract. In the thoracolumbar but not the cervical cord, excitatory commissural connections mediated vestibular responses in contralateral motoneurons. Pharmacological blockade of GABAA receptors showed that responses involved a convergence of excitatory and inhibitory inputs which in combination produced temporal response patterns specific for different segmental levels. Our results show that by birth vestibulospinal projections in rodents have already established functional synapses and are organized to differentially regulate activity in neck and limb motoneurons in a tract- and segment-specific pattern similar to that in adult mammals. Thus, this particular set of descending projections develops several key features of connectivity appropriately at prenatal stages. We also present novel information about vestibulospinal inputs to axial motoneurons in mammals, providing a more comprehensive platform for future studies into the overall organization of vestibulospinal inputs and their role in regulating postural stability.

To better understand how the brainstem reticular formation controls and coordinates trunk and hindlimb muscle activity, we used optical recording to characterize the functional connections between medullary reticulospinal neurons and lumbar motoneurons of the L2 segment in the neonatal mouse. In an isolated brainstem–spinal cord preparation, synaptically induced calcium transients were visualized in individual MNs of the ipsilateral and contralateral medial and lateral motor columns (MMC, LMC) following focal electrical stimulation of the medullary reticular formation (MRF). Stimulation of the MRF elicited differential responses in MMC and LMC, according to a specific spatial organization. Stimulation of the medial MRF elicited responses predominantly in the LMC whereas stimulation of the lateral MRF elicited responses predominantly in the MMC. This reciprocal response pattern was observed on both the ipsilateral and contralateral sides of the spinal cord. To ascertain whether the regions stimulated contained reticulospinal neurons, we retrogradely labelled MRF neurons with axons coursing in different spinal funiculi, and compared the distributions of the labelled neurons to the stimulation sites. We found a large number of retrogradely labelled neurons within regions of the gigantocellularis reticular nucleus (including its pars ventralis and alpha) where most stimulation sites were located. The existence of a mediolateral organization within the MRF, whereby distinct populations of reticulospinal neurons predominantly influence medial or lateral motoneurons, provides an anatomical substrate for the differential control of trunk and hindlimb muscles. Such an organization introduces flexibility in the initiation and coordination of activity in the two sets of muscles that would satisfy many of the functional requirements that arise during postural and non-postural motor control in mammals.

Thalamocortical cells (TCs) and interneurons (INs) in the lateral geniculate nucleus process visual information from the retina. The TCs have many short dendrites, whereas the INs have fewer and longer dendrites. Because of these morphological differences, it has been suggested that transmission of synaptic signals from dendritic synapses to soma is more efficient in TCs than in INs. However, a higher membrane resistance (Rm) for the INs could, in theory, compensate for the attenuating effect of their long dendrites and allow distal synaptic inputs to significantly depolarize the soma. Compartmental models were made from biocytin filled TCs (n= 15) and INs (n= 3) and adjusted to fit the current- and voltage-clamp recordings from the individual cells. The confidence limits for the passive electrical parameters were explored by simulating the influence of noise, morphometric errors and non-uniform and active conductances. One of the useful findings was that Rm was accurately estimated despite realistic levels of active conductance. Simulations to explore the somatic influence of dendritic synapses showed that a small (0.5 nS) excitatory synapse placed at different dendritic positions gave similar somatic potentials in the individual TCs, within the TC population and also between TCs and INs. A linear increase in the conductance of the synapse gave increases in somatic potentials that were more sublinear in INs than TCs. However, when the total synaptic conductance was increased by simultaneously activating many small, spatially distributed synapses, the INs converted the synaptic signals to soma potentials almost as efficiently as the TCs. Thus, INs can transfer fast synaptic signals to soma as efficiently as TCs except when the focal conductance is large.

The basic locomotor rhythm in the cat is generated by a neuronal network in the spinal cord. The exact organization of this network and its drive to the spinal motoneurones is unknown. The purpose of the present study was to use time (cumulant density) and frequency domain (coherence) analysis to examine the organization of the last order drive to motoneurones during fictive locomotion (evoked by application of nialamide and dihydroxyphenylalanine (DOPA)) in the spinal cat. In all cats, narrow central synchronization peaks (half-width < 3 ms) were observed in cumulants estimated between electroneurograms (ENGs) of close synergists, but not between nerves belonging to muscles acting on different joints or to antagonistic muscles. Coherence was not observed at frequencies above 100 Hz and was mainly observed between synergists. Intracellular recording was obtained from a population of 70 lumbar motoneurones. Significant short-term synchronization was observed between the individual intracellular recordings and the ENGs recorded from nerves of the same pool and of close synergists. Recordings from 34 pairs of motoneurones (10 pairs belonged to the same motor pool, 11 pairs to close synergists and 13 pairs to antagonistic pools) failed to reveal any short-lasting synchronization. These data demonstrate that short-term synchronization during fictive locomotion is relatively weak and is restricted to close synergists. In addition, coherence analysis failed to identify any specific rhythmic component in the locomotor drive that could be associated with this synchronization. These results resemble findings obtained during human treadmill walking and imply that the spinal interneurones participating in the generation of the locomotor rhythm are themselves weakly synchronized. The restricted synchronization within the locomotor drive to motoneuronal pools may be a feature of the locomotor generating networks that is related to the ability of these networks to produce highly adaptive patterns of muscle activity during locomotion.

It has been estimated that during locomotion in the cat, muscle afferents in a single limb would produce as many as 800 000 discharges per second (Prochazka & Gorassini, 1998). Such intense sensory feedback could trigger inappropriate reflex responses and detract from processing the most relevant sensory signals during movement. The need to regulate the action of afferent-evoked reflexes was discussed by Eccles & Lundberg (1959b) with regard to the flexion reflex. Under certain experimental conditions, the flexion reflex is evoked by the activation of group II and higher-threshold muscle, as well as joint and cutaneous afferents (Eccles & Lundberg, 1959a). Activation of some of these afferents during the movement could induce disruptive flexion reflexes (Eccles & Lundberg, 1959b). As detectors of static muscle length, group II afferents are active during many movements (see Prochazka et al. 1989). It would thus seem particularly important to modify group II reflex actions and avoid disruptive flexion reflexes during movement (Eccles & Lundberg, 1959b; Grillner & Shik, 1973). Accordingly during brainstem-evoked fictive locomotion, stimulation of certain group II afferents initiates extensor activity instead of evoking flexion reflexes (Perreault et al. 1995). There is evidence that one form of reflex regulation during movement involves a reduction in the central effects of sensory feedback. For example, during voluntary contractions in humans the group Ia monosynaptic excitation of heteronymous motoneurones is reduced at the onset of contraction (Hultborn et al. 1987). Furthermore, our ability to consciously detect afferent signals such as those produced by muscle twitches (Collins et al. 1998) and tactile stimuli (Williams et al. 1998) is decreased during voluntary movement. The inhibitory mechanisms responsible for these reduced sensory effects during voluntary movements are unknown but it has been suggested that a reduction in transmitter release via presynaptic inhibition of primary afferent terminals could play an important role (Hultborn et al. 1987; see Rossignol, 1996). Results from animal experimentation provide further evidence for a presynaptic regulation of primary afferent transmission during the execution of some motor programmes. During fictive locomotion, for example, primary afferents are subject to rhythmic depolarization (Gossard et al. 1989, 1991; Gossard, 1996) and the excitability of the terminals of afferents is increased (Duenas & Rudomin, 1988). Other motor activities in which presynaptic inhibition may modulate excitatory segmental afferent transmission include voiding (Angel et al. 1994; Buss & Shefchyk, 1999), scratching (Bayev & Kostyuk, 1981) and mastication (Kurasawa et al. 1988). The present study used an analysis of monosynaptic field potentials to determine if primary afferent transmission from hindlimb afferents to first-order spinal interneurones is reduced during fictive locomotion. Analysis of monosynaptic field potentials offers several advantages over intracellular recordings in this regard. Because field potentials result from ionic movements, their changes reflect alterations in the currents produced by synaptic transmission between the afferents and their target neurones. On the other hand, intracellular current clamp recordings do not reveal synaptic currents directly, but rather show the postsynaptic voltage changes which are also influenced by postsynaptic membrane conductance and impalement injury. Although all terminals of individual afferents may not be affected uniformly during locomotion, trends in the changes in synaptic transmission may be better appreciated by an analysis of extracellular recorded population responses than by intracellular recordings from single neurones. The strong association between the depression of monosynaptic field potentials and the presence of primary afferent depolarization (see Sypert et al. 1980, for group I afferents, and Riddell et al. 1995, for group II afferents) should aid in comparing results obtained during fictive locomotion and in other preparations. Therefore, to assess the regulation of synaptic transmission from hindlimb afferents during fictive locomotion, afferent-evoked extracellular field potentials were examined during locomotion evoked by stimulation of the mesencephalic locomotor region in decerebrate cats. This preparation has an advantage over drug-induced locomotion because the motor state of the animal can be readily changed from one of quiescence into locomotion. Results will show that there is a general reduction in dorsal and intermediate monosynaptic field potentials evoked by muscle and cutaneous afferents throughout the mid-lumbar to rostral sacral segments examined. This depression is most pronounced for group II field potentials recorded in intermediate spinal laminae. Preliminary results have been reported (Perreault et al. 1994; McCrea & Perreault, 1998).

1. This study examines the effects of electrical stimulation of hindlimb flexor nerves on the fictive locomotion pattern. Locomotion was initiated by stimulation of the mesencephalic locomotor region in the decerebrate paralysed cat and monitored by recording the electroneurogram from selected hindlimb flexor and extensor muscle nerves. Flexor nerves were stimulated using short trains (20-50 stimuli at 100 Hz) during either the flexor or the extensor phase of the fictive locomotor cycle. 2. Stimulation of tibialis anterior (TA), posterior biceps and semitendinosus (PBSt) or sartorius (Sart) nerves at 5 times threshold (T) during the flexor phase of the fictive locomotor cycle terminated on-going activity in flexor nerves and initiated activity in extensors. Thus, flexor nerve stimulation during flexion shortened the locomotor cycle by resetting to extension. The failure of lower intensity (2T) stimulation of PBSt or Sart nerves to reset the step cycle to extension suggests that group II afferents are responsible for these actions. Resetting evoked by 2T stimulation of the TA nerve may be due to a high proportion of group II afferents with low electrical threshold. 3. During extension, stimulation of TA and PBSt nerves at 5T did not perturb the locomotor rhythm whereas Sart stimulation prolonged the locomotor cycle. 4. Stimulation of cutaneous or knee joint afferents failed to produce effects similar to those evoked by stimulation of flexor muscle nerves at group II strength. These findings are at odds with those obtained elsewhere in the acute spinal, DOPA fictive locomotion preparation. The possibility that group II resetting during fictive locomotion is not mediated by flexion reflex pathways but by previously unknown pathways released in the present preparation is discussed. 5. Since many of the flexor afferents recruited by 5T electrical stimulation are the length-sensitive group II fibres, spindle secondaries may act to regulate the duration and onset of flexor and extensor activity during real locomotion. The resetting from flexion to extension also suggests that unexpected or enhanced activity of flexor secondaries during swing would promote a switch of the step cycle to stance.

To assess when vestibulosympathetic projections become functional postnatally, and to establish a preparation in which vestibulosympathetic circuitry can be characterized more precisely, we used an optical approach to record VIIIth nerve-evoked synaptic inputs to thoracic sympathetic preganglionic neurons (SPNs) in newborn mice. Stimulation of the VIIIth nerve was performed in an isolated brainstem–spinal cord preparation after retrogradely labelling with the fluorescent calcium indicator Calcium Green 1-conjugated dextran amine, the SPNs and the somatic motoneurons (MNs) in the thoracic (T) segments T2, 4, 6, 8, 10 and 12. Synaptically mediated calcium responses could be visualized and recorded in individual SPNs and MNs, and analysed with respect to latency, temporal pattern, magnitude and synaptic pharmacology. VIIIth nerve stimulation evoked responses in all SPNs and MNs investigated. The SPN responses had onset latencies from 90 to 200 ms, compared with much shorter latencies in MNs, and were completely abolished by mephenesin, a drug that preferentially reduces polysynaptic over monosynaptic transmission. Bicuculline and picrotoxin, but not strychnine, increased the magnitudes of the SPN responses without changing the onset latencies, suggesting a convergence of concomitant excitatory and inhibitory synaptic inputs. Lesions strategically placed to test the involvement of direct vestibulospinal pathways versus indirect pathways within the brainstem showed that vestibulosympathetic inputs in the neonate are mediated predominantly, if not exclusively, by the latter. Thus, already at birth, synaptic connections in the vestibulosympathetic reflex are functional and require the involvement of the ventrolateral medulla as in adult mammals.

Thalamocortical cells (TCs) and interneurons (INs) in the lateral geniculate nucleus process visual information from the retina. The TCs have many short dendrites, whereas the INs have fewer and longer dendrites. Because of these morphological differences, it has been suggested that transmission of synaptic signals from dendritic synapses to soma is more efficient in TCs than in INs. However, a higher membrane resistance (R(m)) for the INs could, in theory, compensate for the attenuating effect of their long dendrites and allow distal synaptic inputs to significantly depolarize the soma. Compartmental models were made from biocytin filled TCs (n = 15) and INs (n = 3) and adjusted to fit the current- and voltage-clamp recordings from the individual cells. The confidence limits for the passive electrical parameters were explored by simulating the influence of noise, morphometric errors and non-uniform and active conductances. One of the useful findings was that R(m) was accurately estimated despite realistic levels of active conductance. Simulations to explore the somatic influence of dendritic synapses showed that a small (0.5 nS) excitatory synapse placed at different dendritic positions gave similar somatic potentials in the individual TCs, within the TC population and also between TCs and INs. A linear increase in the conductance of the synapse gave increases in somatic potentials that were more sublinear in INs than TCs. However, when the total synaptic conductance was increased by simultaneously activating many small, spatially distributed synapses, the INs converted the synaptic signals to soma potentials almost as efficiently as the TCs. Thus, INs can transfer fast synaptic signals to soma as efficiently as TCs except when the focal conductance is large.