G. N. Orlovsky

Neural networks that co-ordinate locomotion and body orientation in lamprey

The networks of the brainstem and spinal cord that co-ordinate locomotion and body orientation in lamprey are described. The cycle-to-cycle pattern generation of these networks is produced by interacting glutamatergic and glycinergic neurones, with NMDA receptor-channels playing an important role at lower rates of locomotion. The fine tuning of the networks produced by 5-HT, dopamine and GABA systems involves a modulation of Ca2+-dependent K+ channels, high- and low-threshold voltage-activated Ca2+ channels and presynaptic inhibitory mechanisms. Mathematical modelling has been used to explore the capacity of these biological networks. The vestibular control of the body orientation during swimming is exerted via reticulospinal neurones located in different reticular nuclei. These neurones become activated maximally at different angles of tilt.

Activity of vestibulospinal neurons during locomotion

The activity of vestibulospinal neurons giving axons to the lumbosacral spinal cord was recorded during locomotion (walking and running on the treadmill) in mesencephalic and thalamic cats. The overall activity of most neurons increases to a considerable degree during locomotion, and periodic alternations of this activity in relation to the locomotor cycle (modulation) were observed in cats with intact cerebellum. The peak discharge usually occurs at the beginning of the stance phase of the ipsilateral hindlimb, i.e., when the extensor muscles are activated. Phasic modulation disappears when the limbs are stopped by force. There is no modulation in decerebellate cats.

Activity of rubrospinal neurons during locomotion

The activity of rubrospinal neurons giving axons to the lumbosacral spinal cord was recorded during locomotion (walking and running on the treadmill) in thalamic cats. The overall activity of most neurons increased to a considerable degree during locomotion, and strong periodic alternations of this activity in relation to the locomotor cycle (modulation) were observed in cats with intact cerebellum. The peak discharge usually occurred during the swing phase of the contralateral hindlimb, i.e. during the activity of the flexor muscles. Phasic modulation disappeared when the limbs were stopped by force. There was no modulation in decerebellate cats.

Spinal and supraspinal postural networks

Different species maintain a particular body orientation in space (upright in humans, dorsal-side-up in quadrupeds, fish and lamprey) due to the activity of a closed-loop postural control system. We will discuss operation of spinal and supraspinal postural networks studied in a lower vertebrate (lamprey) and in two mammals (rabbit and cat). In the lamprey, the postural control system is driven by vestibular input. The key role in the postural network belongs to the reticulospinal (RS) neurons. Due to vestibular input, deviation from the stabilized body orientation in any (roll, pitch, yaw) plane leads to generation of RS commands, which are sent to the spinal cord and cause postural correction. For each of the planes, there are two groups of RS neurons responding to rotation in the opposite directions; they cause a turn opposite to the initial one. The command transmitted by an individual RS neuron causes the motor response, which contributes to the correction of posture. In each plane, the postural system stabilizes the orientation at which the antagonistic vestibular reflexes compensate for each other. Thus, in lamprey the supraspinal networks play a crucial role in stabilization of body orientation, and the function of the spinal networks is transformation of supraspinal commands into the motor pattern of postural corrections. In terrestrial quadrupeds, the postural system stabilizing the trunk orientation in the transversal plane was analyzed. It consists of two relatively independent sub-systems stabilizing orientation of the anterior and posterior parts of the trunk. They are driven by somatosensory input from limb mechanoreceptors. Each sub-system consists of two closed-loop mechanisms - spinal and spino-supraspinal. Operation of the supraspinal networks was studied by recording the posture-related activity of corticospinal neurons. The postural capacity of spinal networks was evaluated in animals with lesions to the spinal cord. Relative contribution of spinal and supraspinal mechanisms to the stabilization of trunk orientation is discussed.

Control of locomotion in marine mollusc Clione limacina. I. Efferent activity during actual and fictitious swimming.

Summary1.The marine mollusc Clione limacina swims by making rhythmic movements (with a frequency of 1–5 Hz) of its two wings. Filming demonstrated that the wings perform oscillatory movements in the frontal plane of the animal. During both the upward and downward movements of the wing, its posterior edge lagged behind the anterior one, i.e. the wing plane was inclined in relation to the longitudinal axis of an animal. As a result of this inclination, the wing oscillations in the frontal plane produce a force directed forwards. 2.In restrained animals with the body cavity opened (a whole-animal preparation), the wing position, electrical activity in the wing nerve and activity of two identified efferent neurons (1A and 2A) were recorded during locomotory wing movements. There were two bursts of activity in the wing nerve during the locomotory cycle, the first one corresponding to the excitation of efferent neurons controlling the wing elevation, and the second one, to the excitation of efferent neurons controlling the lowering of the wing. Neurons 1A and 2A fired reciprocally at the beginning of the phase of elevating and lowering the wing, respectively. During excitation of one of the neurons, an IPSP appeared in its antagonist. 3. A pair of isolated pedal ganglia of Clione was capable of generating the locomotory rhythm (“fictitious swimming”). In fictitious swimming, as in actual swimming, there were two bursts of activity in the wing nerve per locomotory cycle, and the 1A and 2A neurons fired reciprocally. Homologous neurons from the left and right ganglia fired inphase. A single pedal ganglion was also capable of generating the locomotory rhythm. 4.Serotonin (10-5–10-6 M) increased the locomotor activity both in the whole-animal preparation and in the isolated pedal ganglia.

Control of locomotion in marine mollusc Clione limacina II. Rhythmic neurons of pedal ganglia

Summary1.Activity from neurons in isolated pedal ganglia of Clione limacina was recorded intracellularly during generation of rhythmic swimming. To map the distribution of cells in a ganglion, one of two microelectrodes was used to monitor activity of the identified neuron (1A or 2A), while the second electrode was used to penetrate successively all the visible neurons within a definite area of the ganglion. In addition, pairs of neurons of various types were recorded in different combinations with each other. Intracellular staining of neurons was also performed. 2.Each ganglion contained about 400 neurons, of which about 60 neurons exhibited rhythmic activity related to a swim cycle. These rhythmic neurons were divided into 9 groups (types) according to axonal projections, electrical properties and the phase of activity in a swim cycle. Three types of interneurons and six types of efferent neurons were distinguished. 3.Type 7 and 8 interneurons generated only one spike of long (50–150 ms) duration per swim cycle. Type 7 interneurons discharged in the phase of the cycle that corresponded (in actual swimming) to the dorsal movement of wings (D-phase). Type 8 interneurons discharged in the opposite phase corresponding to the ventral movement of wings (V-phase). With excitation of type 7 interneurons, an IPSP appeared in the type 8 interneurons, and vice versa. Neuropilar branching of these neurons was observed in the ipsilateral ganglion. In addition, they sent an axon to the contralateral ganglion across the pedal commissure. 4.Efferent neurons (i.e. the cell sending axons into the wing nerve) generated spikes of 1–5 ms duration. Type 1 and 3 neurons were excited in the D-phase of a swim cycle and were inhibited in the V-phase. Type 2 and 4 neurons were excited in the V-phase and inhibited in the D-phase. Type 10 neurons received only an excitatory input in the V-phase, while type 6 neurons received only an inhibitory input in the D-phase. 5. Type 12 interneurons were non-spiking cells, they generated a stable depolarization (“plateau”) throughout most of the V-phase. 6. Neurons of the same type from one ganglion (except for type 6) were electrically coupled to each other. There were also electrical connections between most neurons firing in the same phase of the cycle, i.e. between types 3 and 7, as well as between types 2, 4 and 8. Type 7 interneurons from the left and right ganglia were electrically coupled, the same was true for type 8 interneurons.

Activity of Different Classes of Neurons of the Motor Cortex during Postural Corrections

The dorsal side-up body orientation in quadrupeds is maintained by a postural system that is driven by sensory feedback signals. The spinal cord, brainstem, and cerebellum play essential roles in postural control, whereas the role of the forebrain is unclear. In the present study we investigated whether the motor cortex is involved in maintenance of the dorsal side-up body orientation. We recorded activity of neurons in the motor cortex in awake rabbits while animals maintained balance on a platform periodically tilting in the frontal plane. The tilts evoked postural corrections, i.e., extension of the limbs on the side moving down and flexion on the opposite side. Because of these limb movements, rabbits maintained body orientation close to the dorsal side up. Four classes of efferent neurons were studied: descending corticofugal neurons of layer V (CF5s), those of layer VI (CF6s), corticocortical neurons with ipsilateral projection (CCIs), and those with contralateral projection (CCCs). One class of inhibitory interneurons [suspected inhibitory neurons (SINs)] was also investigated. CF5 neurons and SINs were strongly active during postural corrections. In most of these neurons, a clear-cut modulation of discharge in the rhythm of tilting was observed. This finding suggests that the motor cortex is involved in postural control. In contrast to CF5 neurons, other classes of efferent neurons (CCI, CCC, CF6) were much less active during postural corrections. This suggests that corticocortical interactions, both within a hemisphere (mediated by CCIs) and between hemispheres (mediated by CCCs), as well as corticothalamic interactions via CF6 neurons are not essential for motor coordination during postural corrections.

Control of locomotion in marine mollusc Clione limacina III. On the origin of locomotory rhythm

Summary1. Neurons from the isolated pedal ganglia of the marine mollusc Clione limacina were recorded from intracellularly during generation of the locomotory rhythm. Polarization of single type 7 or type 8 interneurons (which discharge in the D-and V-phases of a swim cycle, respectively) strongly affected activity of the rhythm generator. Injection of depolarizing and hyperpolarizing current usually resulted in shortening and lengthening of a swim cycle, respectively. A short pulse of hyperpolarizing current shifted the phase of the rhythmic generator. The same effect could be evoked by polarization of efferent neurons of types 2, 3 and 4 which are electrically coupled to interneurons. On the contrary, polarization of types 1, 6 and 10 efferent neurons, having no electrical connections with interneurons, did not affect the locomotory rhythm. 2. A number of observations indicate that type 7 and 8 interneurons constitute the main source of postsynaptic potentials that were observed in all the “rhythmic” neurons of the pedal ganglia. Type 7 interneurons excited the D-phase neurons and inhibited the V-phase neurons; type 8 interneurons produced opposite effects. 3. Tetrodotoxin eliminated spike generation in all efferent neurons of the pedal ganglia, while in interneurons spike generation persisted. After blocking the spike discharges in all the efferent neurons, type 7 and 8 interneurons were capable of generating alternating activity. One may conclude that these interneurons determine the main features of the swim pattern, i.e., the rhythmic alternating activity of two (D and V) populations of neurons. 4. Both type 7 and type 8 interneurons were capable of endogenous rhythmic discharges with a period like that in normal swimming. This was demonstrated in experiments in which one of the two populations of “rhythmic” neurons (D or V) was inhibited by means of strong electrical hyperpolarization, as well as in experiments in which interaction between the two populations, mediated by chemical synapses, was blocked by Co2+ ions. 5. Type 7 and 8 interneurons were capable of “rebound”, i.e. they had a tendency to discharge after termination of inhibition. 6. V-phase neurons exerted not only inhibitory but also excitatory action upon D-phase neurons, the excitatory action being longer than the inhibitory one. 7. The main experimental findings correspond well to the model of rhythm generator consisting of two half centres possessing endogenous rhythmic activity. The half-centres exert strong, short duration inhibitory and weak long duration excitatory actions upon one another. The behaviour of such a model is considered and compared with that of the locomotor generator of Clione.

Activity of Ia inhibitory interneurons during fictitious scratch reflex in the cat.

(1) The fictitious scratch reflex was observed in decerebrate cats immobilized with Flaxedil. The activity of interneurons in the inhibitory pathways from Ia afferents to motoneurons of antagonistic muscles was recorded during scratching. The selected interneurons were supplied by Ia afferents from m. vastus, posterior biceps-semitendinosus and sartorius. (2) Almost all recorded interneurons showed periodic modulation of activity. Their maximal activity in the scratch cycle usually coincided with the maximal activity of motoneurons of those muscles from which the interneurons receive Ia afferents. Excitation of Ia afferents by passive stretch of the muscle or by electrical stimulation of the muscle nerve resulted in the increase of the interneuron activity, without changing its timing in the scratch cycle.

Encoding and decoding of reticulospinal commands.

In the lamprey, the reticulospinal (RS) system is the main descending system transmitting commands to the spinal cord. We investigated these commands and their effect on the spinal mechanisms. The RS commands were studied by recording responses of RS neurons to sensory stimuli eliciting different motor behaviors. Initiation of locomotion was associated with symmetrical bilateral massive activation of RS neurons, whereas turns in different planes were associated with asymmetrical activation of corresponding neuronal groups. The sub-populations of RS neurons causing different motor behaviors partly overlap. We suggest that commands for initiation of locomotion and regulation of its vigour, encoded as the value of bilateral RS activity, are decoded in the spinal cord by integrating all RS signals arriving at the segmental locomotor networks. Commands for turns in different planes, encoded as an asymmetry in the activities of specific groups of RS neurons, are decoded by comparing the activities of those groups. This hypothesis was supported by the experiments on a neuro-mechanical model, where the difference between the activities in the left and right RS pathways was used to control a motor rotating the animal in the roll plane. Transformation of the descending commands into the motor responses was investigated by recording the effects of individual RS neurons on the motor output. Twenty patterns of influences have been found. This great diversity of the patterns allows the RS system to evoke body flexion in any plane. Since most neurons have asymmetrical projections we suggest that, for rectilinear swimming, RS neurons with opposite asymmetrical effects are co-activated.