Cerebellar Anterior Vermis Research Articles

Cerebellar Metabolic Connectivity during Treadmill Walking before and after Unilateral Dopamine Depletion in Rats.

Compensatory changes in brain connectivity keep motor symptoms mild in prodromal Parkinson's disease. Studying compensation in patients is hampered by the steady progression of the disease and a lack of individual baseline controls. Furthermore, combining fMRI with walking is intricate. We therefore used a seed-based metabolic connectivity analysis based on 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) uptake in a unilateral 6-OHDA rat model. At baseline and in the chronic phase 6-7 months after lesion, rats received an intraperitoneal injection of [18F]FDG and spent 50 min walking on a horizontal treadmill, followed by a brain PET-scan under anesthesia. High activity was found in the cerebellar anterior vermis in both conditions. At baseline, the anterior vermis showed hardly any stable connections to the rest of the brain. The (future) ipsilesional cerebellar hemisphere was not particularly active during walking but was extensively connected to many brain areas. After unilateral dopamine depletion, rats still walked normally without obvious impairments. The ipsilesional cerebellar hemisphere increased its activity, but narrowed its connections down to the vestibulocerebellum, probably aiding lateral stability. The anterior vermis established a network involving the motor cortex, hippocampus and thalamus. Adding those regions to the vermis network of (previously) automatic control of locomotion suggests that after unilateral dopamine depletion considerable conscious and cognitive effort has to be provided to achieve stable walking.

Resting-state perfusion in motor and fronto-limbic areas is linked to diminished expression of emotion and speech in schizophrenia

Negative symptoms (NS) are a core component of schizophrenia affecting community functioning and quality of life. We tested neural correlates of NS considering NS factors and consensus subdomains. We assessed NS using the Clinical Assessment Interview for Negative Symptoms and the Scale for Assessment of Negative Symptoms. Arterial spin labeling was applied to measure resting-state cerebral blood flow (rCBF) in 47 schizophrenia patients and 44 healthy controls. Multiple regression analyses calculated the relationship between rCBF and NS severity. We found an association between diminished expression (DE) and brain perfusion within the cerebellar anterior lobe and vermis, and the pre-, and supplementary motor area. Blunted affect was linked to fusiform gyrus and alogia to fronto-striatal rCBF. In contrast, motivation and pleasure was not associated with rCBF. These results highlight the key role of motor areas for DE. Considering NS factors and consensus subdomains may help identifying specific pathophysiological pathways of NS.

Effects of trunk-to-head rotation on the labyrinthine responses of rat reticular neurons

Vestibulospinal reflexes elicited by head displacement become appropriate for body stabilization owing to the integration of neck input by the cerebellar anterior vermis. Due to this integration, the preferred direction of spinal motoneurons’ responses to animal tilt rotates by the same angle and by the same direction as the head over the body, which makes it dependent on the direction of body displacement rather than on head displacement. It is known that the cerebellar control of spinal motoneurons involves the reticular formation. Since the preferred directions of corticocerebellar units’ responses to animal tilt are tuned by neck rotation, as occuring in spinal motoneurons, we investigated whether a similar tuning can be observed also in the intermediate station of reticular formation. In anaesthetized rats, the activity of neurons in the medullary reticular formation was recorded during wobble of the whole animal at 0.156Hz, a stimulus that tilted the animal’s head by a constant amplitude (5°), in a direction rotating clockwise or counter clockwise over the horizontal plane. The response gain and the direction of tilt eliciting the maximal activity were evaluated with the head and body axes aligned and during a maintained body-to-head displacement of 5–20° over the horizontal plane, in either direction. We found that the neck displacement modified the response gain and/or the average activity of most of the responsive neurons. Rotation of the response direction was observed only in a minor percentage of the recorded neurons. The modifications of reticular neurons’ responses were different from those observed in the P-cells of the cerebellar anterior vermis, which rarely showed gain and activity changes and often exhibited a rotation of their response directions. In conclusion, reticular neurons take part in the neck tuning of vestibulospinal reflexes by transforming a head-driven sensory input into a body-centred postural response. The present findings prompt re-evaluation of the role played by the reticular neurons and the cerebellum in vestibulospinal reflexes.

Effects of Leg-to-Body Position on the Responses of Rat Cerebellar and Vestibular Nuclear Neurons to Labyrinthine Stimulation

The spatial organization of vestibulospinal (VS) reflexes, elicited by labyrinthine signals and related to head motion, depends on the direction of body tilt, due to proprioceptive neck afferents acting through the cerebellar anterior vermis. The responses of Purkinje cells located within this region to labyrinthine stimulation are modulated by the head-to-body position. We investigated, in urethane-anesthetized rats, whether a 90° leg-to-trunk displacement modifies the responses of corticocerebellar and vestibular nuclear neurons to the labyrinthine input, which would indicate that VS reflexes are tuned by the leg-to-trunk position. With this aim, unit activity was recorded during "wobble" stimuli that allow evaluating the gain and spatiotemporal properties of neuronal responses. The response gain of corticocerebellar units showed a significant drop in the leg-rotated position with respect to the control one. Following a change in leg position, a proportion of the recorded neurons showed significant changes in the direction and phase of the response vector. In contrast, vestibular nuclear neurons did not show significant modifications in their response gain and direction. Thus, proprioceptive afferents signaling leg-to-trunk position seem to affect the processing of directional labyrinthine signals within the cerebellar cortex.

Responses of Purkinje-cells of the cerebellar anterior vermis to stimulation of vestibular and somatosensory receptors

In decerebrate cats, sinusoidal rotation of the forepaw around the wrist modifies the activity of the ipsilateral forelimb extensor triceps brachii (TB) and leads to plastic changes of adaptive nature in the gain of vestibulospinal (VS) reflexes (VSRs). Both effects are depressed by functional inactivation of the cerebellar anterior vermis, which also decreases the gain of VSRs. In order to better understand the mechanisms of these phenomena, the simple spike activity of Purkinje (P-) cells was recorded from the vermal cortex of the cerebellar anterior lobe during individual and/or combined stimulation of somatosensory wrist, neck and vestibular receptors. About one third of the recorded units were affected by sinusoidal rotation of the ipsilateral forepaw around the wrist axis (0.16 Hz, ±10°). Most of these neurons (∼60%) increased their activity during ventral flexion of the wrist and decreased it during the oppositely directed movement, with an average phase lag of −141° with respect to the position of maximal dorsiflexion. The remaining cells (∼40%) were excited during dorsiflexion of the wrist, with an average phase lead of 59° with respect to the extreme dorsal flexion. Both populations showed comparable response gains, with an average value of 0.42±0.52, S.D., imp/s/deg. About half of the recorded units were also tested during sinusoidal roll tilt of the animal around the longitudinal axis (0.16 Hz, ±10°), leading to stimulation of labyrinthine receptors. When both stimuli were applied simultaneously, the responses to combined stimulation usually corresponded to the sum of individual responses. While the phase distribution of somatosensory responses was clearly bimodal, vestibular responses showed phase angle values uniformly scattered between ±180° and 0°, so that, during combined stimulation, each neuron could be maximally activated by coupling the two stimuli with a particular phase relation. Finally, a proportion of the recorded neurons was also tested during sinusoidal rotation of the body around its longitudinal axis, with the head fixed in space, leading to stimulation of neck receptors. The proportion of neurons affected by individual stimulation of vestibular or neck receptors (81% and 72%, respectively) was larger than that of wrist-driven neurons. Convergence of signals from vestibular, somatosensory wrist and neck receptors was found in 18% of the neurons analyzed. In conclusion, the results of this study show that somatosensory signals from the forelimb: i) modulate the activity of a sizeable proportion of neurons located within the cerebellar anterior vermis and ii) interact widely with labyrinthine and neck signals at this level. Moreover, iii) this corticocerebellar region is largely dominated by vestibular and neck signals that may be utilized to build up a neuronal representation of the position of body in space. These findings suggest that: 1) the modulation of TB activity induced by rotation of the ipsilateral wrist may at least partially depend upon the simultaneous changes in P-cell activity and 2) the interaction of vestibular and somatosensory wrist signals at P-cell level may represent the substrate of the plastic changes that affect the VSR when animal tilt and wrist rotation are driven together. A preliminary report of these data has been presented [ Bruschini L, Manzoni D, Pompeiano O (2000) Responses of cerebellar Purkinje cells to forepaw rotation in decerebrate cat. Pflügers Arch 440:R31].

Proprioceptive neck influences modify the information about tilt direction coded by the cerebellar anterior vermis

ObjectiveTo verify whether the direction of head tilt coded by the population response of Purkinje (P) cells located in the cerebellar anterior vermis is modified by the relative position of the body with respect to the head. Material and MethodsIn decerebrate cats, the responses of P cells to wobble of the whole animal were analyzed in order to compute the response vectors of the same cells to the labyrinthine input. These response vectors were used to evaluate the population response (vector) of all the recorded neurons to head tilt in specific directions. ResultsWhen the head was aligned with the body, the direction of the population vector closely corresponded to that of head tilt. A 30° body-to-head rotation to the left or right around a C1–C2 vertical axis modified the response vectors of most of the recorded neurons, leading to reciprocal deviations of the population vector from the direction of head tilt of ≈30°. ConclusionWe propose that information from neck receptors regulates the convergence of labyrinthine signals with different spatial and temporal properties on corticocerebellar units, thus allowing the P-cell population to code the direction of body tilt.

Postural responses of forelimb extensors to somatosensory signals elicited during wrist rotation: interaction with vestibular reflexes.

The responses of the forelimb extensor triceps brachii (TB) to sinusoidal wrist rotation, at 0.156 Hz, +/-5 degrees, were investigated in decerebrate cats. In most of the tested muscles [12/15) the multiunit electromyographic (EMG) activity of the TB muscle increased during dorsiflexion and decreased during plantar flexion of the ipsilateral limb extremity, showing an average (+/-SD) gain of 1.38+/-0.64 impulses/s/ degrees and a phase lead of 33+/-19 degrees with respect to the extreme dorsiflexion of the forepaw. Both parameters remained unmodified by increasing the amplitude of stimulation from 5 degrees up to 10 degrees. Rotation of the contralateral wrist had no effect on TB activity. When the rotation of the ipsilateral wrist occurred during sinusoidal roll tilt of the whole animal, a stimulus that activates vestibular receptors, the TB response to combined stimulation closely corresponded to the vectorial sum of the individual responses. Dorsiflexion of the forepaw could drive the vestibulospinal system, which excites the ipsilateral TB motoneurons. In fact, functional inactivation of the cerebellar anterior vermis, which controls vestibulospinal neurons, significantly modified the TB response to wrist rotation. It appears, therefore, that the somatosensory signals elicited by wrist rotation utilize the spinocerebellum to modulate the activity of the TB. The TB responses to wrist rotation could play a role in stabilizing posture during stance and locomotion.

Behavioral abnormalities of Zic1 and Zic2 mutant mice: implications as models for human neurological disorders.

Zic1 and Zic2 encode closely related zinc finger proteins expressed in dorsal neural tube and its derivatives. In previous studies, we showed that the homozygous Zic1 null mutation (Zic1-/-) results in cerebellar malformation with severe ataxia and that holoprosencephaly and spina bifida occur in homozygotes for Zic2 knockdown mutation (Zic2kd/kd). Since human ZIC2 haploinsufficiency is a cause of holoprosencephaly, the Zic2kd/kd mice are regarded as an animal model for holoprosencephaly in humans. In this study, the behavioral characteristics of the Zic1 and Zic2 mutant mice were investigated in heterozygotes (Zic1-/+ or Zic2kd/+), and significant abnormalities were found in the hanging, spontaneous locomotor activity, stationary rod (Zic1-/+), acoustic startle response, and prepulse inhibition tests (Zic2kd/+). The abnormalities in the Zic1-/+ mice may be explained in part by the hypotonia caused by hypoplasia of the cerebellar anterior vermis, and these mice are regarded as a model of Joubert syndrome. In contrast, the sensorimotor gating abnormality in the Zic2kd/+ mice may be attributable to the presumed abnormality in the dorsomedial forebrain, which was strongly affected in the Zic2kd/kd mice. Zic2kd/+ mice can serve as a model for diseases involving sensorimotor gating abnormalities, such as schizophrenia.

Neck input modifies the reference frame for coding labyrinthine signals in the cerebellar vermis: a cellular analysis

The activity of 68 neurons, mainly Purkinje cells, was recorded from the cerebellar anterior vermis of decerebrate cats during wobble of the whole animal (at 0.156 Hz, 5°), a mixture of tilt and rotation, leading to stimulation of labyrinth receptors. Most of the neurons (65/68) were affected by both clockwise and counterclockwise rotations. Twenty-four units showing responses of comparable amplitude to these stimuli (narrowly tuned cells) were represented by a single vector (S max), whose preferred direction corresponded to the direction of stimulation giving rise to the maximal response. The remaining 41 units, however, showed different amplitude responses to these rotations (broadly tuned cells) and were characterized by two spatially and temporally orthogonal vectors (S max and S min), suggesting that labyrinthine signals with different spatial and temporal properties converged on these cells. All these units were tested while the body was aligned with the head (control position), as well as after static displacement of the body under a fixed head by 15° and/or 30° around a vertical axis passing through C1–C2, thus leading to stimulation of neck receptors. The orientation component of the response vector of the Purkinje cells to vestibular stimulation changed following body-to-head displacement. Moreover, the amplitude of vector rotation corresponded, on the average, to that of body rotation. Changes in temporal phase, gain and tuning ratio of the responses were also observed. We propose that information from neck receptors regulates the convergence of labyrinthine signals with different spatial and temporal properties on corticocerebellar units. Due to their strict relationship with the motor system, these units may give rise to appropriate responses in the limb musculature, by modifying the spatial organization of the vestibulospinal reflexes according to the requirements of body stability. The cerebellar vermis may thus represent an important structure, where frames of reference can be altered to account for changes in position of trunk, head and neck.

Brain activation during maintenance of standing postures in humans.

The regulatory mechanism of bipedal standing in humans remains to be elucidated. We investigated neural substrates for maintaining standing postures in humans using PET with our mobile gantry PET system. Normal volunteers were instructed to adopt several postures: supine with eyes open toward a target; standing with feet together and eyes open or eyes closed; and standing on one foot or with two feet in a tandem relationship with eyes open toward the target. Compared with the supine posture, standing with feet together activated the cerebellar anterior lobe and the right visual cortex (Brodmann area 18/19), and standing on one foot increased cerebral blood flow in the cerebellar anterior vermis and the posterior lobe lateral cortex ipsilateral to the weight-bearing side. Standing in tandem was accompanied by activation within the visual association cortex, the anterior and posterior vermis as well as within the midbrain. Standing with eyes closed activated the prefrontal cortex (Brodmann area 8/9). Our findings confirmed that the cerebellar vermis efferent system plays an important role in maintenance of standing posture and suggested that the visual association cortex may subserve regulating postural equilibrium while standing.

Neck Influences on the Spatial Properties of Vestibulospinal Reflexes in Decerebrate Cats: Role of the Cerebellar Anterior Vermis

The vestibulospinal (VS) reflexes elicited by animal rotation modify the activity of limb musculature, thus preserving balance and postural stability. We investigated whether the orientation of these postural responses is strictly dependent upon the direction of head displacement or else can be modified by extralabyrinthine inputs to the goal of stabilizing body position. The experiments were performed in decerebrate cats, in which the effects of static body-to-head displacements were tested on the multiunit EMG responses of the medial head of the triceps brachii to wobble of the whole animal at 0.15 Hz, 10 degrees, both in the clockwise (CW) and counterclockwise (CCW) direction. These stimuli allowed us to determine the muscle response vector, whose orientation component corresponds to the direction of head displacement giving rise to the maximal EMG response. When the animal body was kept straight with respect to the head, the triceps response vector was always oriented close to the transverse axis, pointing to the side-down direction. Following 30 degrees of body-to-head displacement around a vertical axis passing through the first-second cervical joints, the response vectors of both the left and the right muscles shifted in the same direction of body rotation, thus remaining approximately perpendicular to the body axis. The change in muscle vector orientation corresponded on the average to the angle of body-to-head displacement. Only slight changes in amplitude of the muscle responses were observed. These findings imply that the maximal activation of the triceps brachii always occurred for the same direction of body displacement, irrespective of the pattern of discharge of vestibular afferents, which is determined by the direction of head displacement. The rotation of the triceps response vector induced by body-to-head displacement was reduced or suppressed by inactivation of the ipsilateral cerebellar anterior vermis, following local microinjection of the GABA(A) agonist muscimol. These findings indicate that 1) the sensory input which results from changing the body position with respect to the head, probably originating from neck receptors, is able to modify the pattern of the VS reflexes, which appear to be organized in a body-centered reference frame, and 2) the cerebellar vermis is required for the proper execution of this sensorimotor transformation.

Noradrenergic influences on the cerebellar cortex: Effects on vestibular reflexes under basic and adaptive conditions

Experiments performed either in decerebrate cats or in intact rabbits have shown that functional inactivation of the cerebellar anterior vermis or the flocculus decreased the basic gain of the vestibulospinal or the vestibulo-ocular reflex, respectively. These findings were attributed to the fact that a proportion of the vermal or floccular Purkinje cells, which are inhibitory in function, discharge out of phase with respect to the vestibulospinal or the vestibulo-ocular neurons during sinusoidal animal rotation, thus exerting a facilitatory influence on the gain of the vestibular reflexes. Intravermal injection of a β-noradrenergic agonist slightly increased the gain of the vestibulospinal reflex, whereas the opposite result was obtained after injection of β-antagonists. Similarly, intrafloccular injection of a β-noradrenergic agonist slightly facilitated the gain of the vestibulo-ocular reflex in darkness (but not in light), whereas a small decrease of the reflex occurred after injection of a β-antagonist. It was postulated that the noradrenergic system acts on Purkinje cells by enhancing their amplitude of modulation to a given labyrinth signal, thus increasing the basic gain of the vestibular reflexes. The Purkinje cells of the cerebellar anterior vermis and the flocculus also exert a prominent role on the adaptation of vestibulospinal and vestibulo-ocular reflexes, respectively. In particular, intravermal or intrafloccular injection of β-noradrenergic antagonists decreased or suppressed the adaptive capacity of the vestibulospinal and vestibulo-ocular reflexes that always occurred during sustained out-of-phase neck-vestibular or visual-vestibular stimulation, whereas the opposite result was obtained after local injection of a β-noradrenergic agonist. The noradrenergic innervation of the cerebellar cortex originates from the locus coeruleus complex, whose neurons respond to vestibular, neck, and visual signals. It was postulated that this structure acts through β-adrenoceptors to increase the expression of immediate-early genes, such as c- fos and Jun-B, in the Purkinje cells during vestibular adaptation. Induction of immediate-early genes could then represent a mechanism by which impulses elicited by sustained neck-vestibular or visuovestibular stimulation are transduced into long-term biochemical changes that are required for cerebellar long-term plasticity. (Otolaryngol Head Neck Surg 1998;119:93-105.)

Spatiotemporal response properties of cerebellar Purkinje cells to neck displacement

The activity of 184 Purkinje cells and 58 unidentified neurons located within the cerebellar anterior vermis was recorded in decerebrate cats during wobble of the body under a fixed head. This stimulus induced a neck displacement of constant amplitude (2.5°) whose direction rotated at the constant velocity of 56.2°/s on the horizontal plane, both in the clockwise and counterclockwise directions. It was then possible to evaluate the spatiotemporal characteristics of unit responses to neck displacement in the vertical planes; 131 of 184 Purkinje cells (71%) and 35 of 58 unidentified cells (60%) responded to clockwise and/or counterclockwise rotations. In particular, among the responsive units, 44% of the Purkinje cells and 37% of the unidentified cells showed an equal amplitude modulation during clockwise and counterclockwise rotations. These units are expected to show a maximal response sensitivity for neck displacement in a preferred direction, a null response for perpendicularly oriented stimuli and a constant temporal phase (narrowly tuned neurons). In 28% of the Purkinje cells and 40% of the unidentified cells, responses of different amplitudes were observed during clockwise and counterclockwise rotations. These neurons should display a preferred direction of response to neck displacement, lack of null response directions and a temporal phase changing with the stimulus direction (broadly tuned neurons). Finally, 27% of the Purkinje cells and 23% of the unidentified cells responded only to wobble in the clockwise or counterclockwise direction (unidirectional units). This behavior predicts equal sensitivities for all the directions of neck displacement and a response phase changing linearly with the direction of neck displacement. A maximal sensitivity vector ( S max), aligned with the preferred direction of the neuron, was evaluated for the bidirectional narrowly tuned and broadly tuned units. Its amplitude and temporal phase corresponded to the response characteristics expected for stimuli in the preferred direction of the cell. S max directions were distributed over the horizontal plane. Most of them, however, were closer to the pitch than to the roll axis and pointed towards the animal's tail. Among pitch-related Purkinje cells, the temporal phase of S max was small with a predominance of phase lags; phase leads of rather large amplitude were usually observed for roll-related Purkinje cells. The possibility that the recorded population of units coded the direction of neck displacement was tested by assuming that each cell gave a vectorial contribution related to its response properties and that the vectorial sum of such contributions represented the outcome of the population code. Dynamic body-to-head displacements in four different directions were simulated and for each direction 12 population vectors were evaluated at regular intervals of the stimulus cycle. The direction of the population vector was related to that of the stimulus, but the correspondence was close only for the pitch direction. Moreover, the amplitude of the population vector depended upon the direction of the stimulus, being larger for pitch than for roll displacements. Due to the efferent connections of the explored cerebellar region, the neuronal signals generated by the Purkinje cell population are probably transferred to the spinal cord, where they may differentially affect the amplitude and the spatial properties of the neck reflexes according to the direction of neck displacement.

Convergence of directional vestibular and neck signals on cerebellar purkinje cells.

Convergence of spatially oriented vestibular and neck signals within the cerebellar anterior vermis in decerebrate cats was studied by recording the simple spike discharge of Purkinje (P) cells during wobble either of the whole animal (vestibular input) or of the body under a fixed head (neck input) at 0.156 Hz, 5 degrees and 2.5 degrees , respectively. Both clockwise (CW) and counterclockwise (CCW) rotations were performed. Units that had equal response amplitudes to CW and CCW rotations (narrowly tuned neurons) were described by a single vector (Smax), characterized by a gain, a direction and a temporal phase. Units with different response amplitudes to CW and CCW rotation (broadly tuned neurons) were described by two vectors (Smax and Smin). In addition to these bidirectional units, there were also unidirectional units which responded either to CW or CCW rotation; in these cases the gain of Smax equals that of Smin. On the whole, 77% and 63% of the P cells responding to vestibular and neck stimulation, respectively, showed a bidirectional broadly tuned or unidirectional behavior. These response patterns were attributed to the convergence of signals with different spatial and temporal properties. About 50% of the P cells from which recordings were made responded to stimulation of both sensory systems. However, the gains of the Smax vectors of the neck responses were much greater than those of the vestibular responses, at least for small amplitudes of rotation, and were positively correlated with them. Usually the differences in the orientation components of the neck and vestibular Smax vectors were larger, while the differences in temporal phases were smaller than 90 degrees . These findings suggest that periodic changes in the phase difference and gain ratio of the neck to the vestibular response may occur during dynamic displacement of the head over the body, depending on the stimulus direction. As a result of these data, the P cells of the cerebellar vermis are expected to show prominent responses to head rotation, which could affect the spatially organized postural responses by utilizing vestibular and reticular targets.

Spatiotemporal response properties of cerebellar Purkinje cells to animal displacement: a population analysis

The hypothesis that corticocerebellar units projecting to vestibulospinal neurons contribute to the spatiotemporal response characteristics of forelimb extensors to animal displacement was tested in decerebrate cats in which the activity of Purkinje cells and unidentified cells located in the cerebellar anterior vermis was recorded during wobble of the whole animal. This stimulus imposed to the animal a tilt of fixed amplitude (5°) 1 This stimulus amplitude was used since, in a previous study, [18]the responses to a roll tilt of a large population of P-cells located in the vermal cortex of the cerebellar anterior lobe showed a linearity in the range of 2–15°. 1 , with a direction moving at a constant angular velocity (56.2°/s), both in the clockwise and counterclockwise directions over the horizontal plane. Eighty-three percent (143/173) of Purkinje cells and 81% (42/52) of unidentified cells responded to clockwise and/or counterclockwise rotations. In particular, 116/143 Purkinje cells (81%) and 32/42 unidentified cells (76%) responded to both clockwise and counterclockwise rotations (bidirectional units), while 27/143 Purkinje cells (19%) and 10/42 unidentified cells (24%) responded to wobble in one direction only (unidirectional units). For the bidirectional units, the direction of maximum sensitivity to tilt ( S max) was identified. Among these units, 24% of the Purkinje cells and 26% of the unidentified cells displayed an equal amplitude of modulation during clockwise and counterclockwise rotations, indicating a cosine-tuned behavior. For this unit type, the temporal phase of the response to a given direction of tilt should remain constant, while the sensitivity would be maximal along the S max direction, declining with the cosine of the angle between S max and the tilt direction. The remaining bidirectional units, i.e. 57% of the Purkinje cells and 50% of the unidentified cells displayed unequal amplitudes of modulation during clockwise and counterclockwise rotations. For these neurons, a non-zero sensitivity along the null direction is expected, with a response phase varying as a function of stimulus direction. As to the unidirectional units, their responses to wobble in one direction predict equal sensitivities along any tilt direction in the horizontal plane and a response phase that changes linearly with the stimulus direction. By comparing these data with those obtained previously during selective stimulation of macular receptors by a 5° off-vertical axis rotation, it appeared that the directions of maximum sensitivity to tilt were distributed over the whole horizontal plane of stimulation, in both conditions. However, co-stimulation of macular and canal receptors during wobble decreased the proportion of unidirectional units, while that of the bidirectional, namely broadly tuned units, increased. In addition, while the average gain of the S max vector of the bidirectional units was comparable, the temporal phase of this vector tended to show a more prominent phase leading behavior during wobble with respect to off-vertical axis rotation. The possibility that the tested neurons formed a population which coded the direction of head tilt in space was also investigated using a modified version of the classical population vector analysis. It was shown that for each selected time in the tilt cycle the direction of the population vector closely corresponded to that of the head tilt, while its amplitude was related to that of the stimulus. We conclude that the broad distribution of the response vector orientation of units located in the cerebellar anterior vermis represents an appropriate substrate for the cerebellar control of vestibulospinal reflexes involving extensor muscles during a variety of head tilts. In view of their efferent projections to the vestibular and fastigial nuclei, the cerebellar anterior vermis may provide a framework for the spatial coding of vestibular inputs, giving equal emphasis to both side-to-side and fore–aft stability.

Changes in Gain and Spatiotemporal Properties of the Vestibulospinal Reflex after Injection of a GABA-A Agonist in the Cerebellar Anterior Vermis

Experiments were performed to study the influence of the cerebellar anterior vermis on both amplitude and directional properties of the vestibulospinal (VS) reflexes. In decerebrate cats, the multiunit EMG activity of the medial head of the forelimb extensor triceps brachii was recorded during wobble of the whole animal at 0.15 Hz. With this procedure the animals were submitted to a tilt characterized by a fixed amplitude (10 degrees) and by a direction moving at constant velocity over the horizontal plane, in both a clockwise (CW) and a counterclockwise (CCW) direction. These dynamic stimuli permitted characterization of the triceps muscle response to animal tilt as a single vector in the horizontal plane. The gain of this vector was taken as the mean value obtained for the CW and CCW responses, while its orientation corresponded to the direction of head displacement, lying midway between the maximal response directions to CW and CCW rotations. The temporal phase was evaluated as the half difference between the directions of the CW and CCW responses. In all the experiments the response vector of the triceps brachii was closely aligned with the transverse axis and pointed to the side-down direction. Unilateral inactivation of the cerebellar anterior vermis after microinjection, in one or two folia of lobule V, of the GABA-A agonist muscimol (0.5 microL at 8 micrograms/microL saline), consistently and reversibly reduced in 20 to 40 min the amplitude of the EMG modulation of the ipsilateral triceps brachii to 46% to 80% of the control value, while only a small shift (up to 30 degrees) of the response vector occurred either nosewards or tailwards. Small shifts in temporal phase were also observed. These findings suggest that the Purkinje (P)-cells, which usually fire out of phase with respect to the VS neurons, contribute positively to the amplitude of the VS reflexes. It was previously shown that P-cells with response vectors covering all the directions of animal displacement are present in small regions of the cerebellar anterior vermis; it is likely that these neurons represent functional units facilitating the VS reflexes elicited by animal tilt in the direction of their response vectors. By suppressing the activity of these cells, muscimol injections would lead to a general depression of the triceps responses to animal displacement, not associated with prominent changes in directional specificity.

Responses of Purkinje cells in the cerebellar anterior vermis to off-vertical axis rotation.

Responses of 67 Purkinje cells (P-cells) and 44 unidentified neurons (U-cells) located in the cerebellar anterior vermis were recorded in decerebrate cats during off-vertical axis rotation (OVAR). This stimulus consisted of a slow constant velocity (9.4%/s) rotation in the clockwise (CW) and counterclockwise (CCW) directions around an axis inclined by 5 degrees with respect to the vertical. OVAR imposes on the animal head a 5 degrees tilt, whose direction changes continuously over the horizontal plane, thus eliciting a selective stimulation of macular receptors. A total of 27/67 P-cells (40%) and 24/44 U-cells (55%) responded to both CW and CCW rotations. For these bidirectional units, the direction of maximum sensitivity to tilt (Smax) could be identified. Smax directions were distributed over the whole horizontal plane of stimulation. Among bidirectional neurons, 48% of the P-cells and 33% of the U-cells displayed an equal amplitude of modulation during CW and CCW rotations, indicating a cosine-tuned behaviour. In these instances, the temporal phase of the unit response to a given direction of tilt remained constant, while the sensitivity was maximal along the Smax direction and declined with the cosine of the angle between Smax and the tilt direction. The remaining bidirectional units displayed unequal amplitudes of modulation during CW and CCW rotations. For these neurons, a nonzero sensitivity along the null direction was expected and the response phase varied as a function of stimulus direction. Finally, 31% and 23% of P-cells and U-cells, respectively, responded during OVAR in one direction only (unidirectional units). This behaviour predicts equal sensitivities along any tilt direction in the horizontal plane and a response phase that changes linearly with the stimulus direction. The posibility that the tested neurons formed a population which coded the direction of head tilt in space was also investigated. The data from the whole population of cells were analysed using a modified version of vectorial analysis. This model assumes that for a particular tilt each cell makes vectorial contributions; the vectorial sum of these contributions represent the outcome of the population code and points in the direction of head tilt in space. Thus, a dynamic head tilt along four representative directions was simulated. For each of the four directions, 12 population vectors were calculated at regular time intervals so as to cover an entire cycle of head tilt. The results indicate that for each selected time in the cycle the direction of the population vector closely corresponded to that of the head tilt, while its amplitude was related to the amount of head tilt. These data were particularly obtained for the P-cells. In view of their efferent connections, the cerebellar anterior vermis may provide a framework for the spatial organization of vestibulospinal reflexes induced by stimulation of otolith receptors.

Role of the spinocerebellum in adaptive gain control of cat's vestibulospinal reflex.

In decerebrate cats, a 3-h period of sustained roll tilt of the head (at 0.15 Hz. +/- 10) leading to selective stimulation of labyrinth receptors, associated with a synchronous roll tilt of the body (at 0.15 Hz., +/- 12.5) leading to 2.5 degrees out-of-phase neck rotation produced an adaptive increase in gain of the vestibulospinal reflex (VSR) elicited by roll tilt of the animal at 0.15 Hz, +/- 10 degrees. This increase reached the maximum at the end of the third h of stimulation and persisted unmodified during the first h after stimulation. Microinjection into zone B of the cerebellar anterior vermis of the GABA-A agonist muscimol (0.25 microliter at 8 micrograms/microliters saline), producing only a slight or negligible depression of the VRS gain in non-adaptive conditions, prevented the occurrence of the adapted increase in gain of the VSR following a 3-h period of sustained head-body rotation. Moreover, intravermal injection of the GABA-A agonist muscimol or the GABA-B agonist baclofen (0.25 microliter at 8 or 2 micrograms/microliters saline, respectively) suppressed the already adapted VSR gain. It is postulated that the adaptive increase in gain of the VSR following a sustained neck-vestibular stimulation depends on plastic changes which affect the Purkinje cells of the cerebellar anterior vermis.

Role of muscarinic receptors in the cerebellar control of the vestibulospinal reflex gain: cellular mechanisms.

Most of the inhibitory Purkinje (P-) cells of the cerebellar anterior vermis fire out-of-phase with respect to the excitatory vestibulospinal neurons during roll tilt of the animal, thus exerting a positive influence on the gain of the vestibulospinal reflex (VSR). The responses of these P-cells depend on activation of glutamatergic excitatory mossy fibers-granule cells, but they are likely to be shaped by GABAergic inhibitory interneurons. The cerebellar cortex contains cholinergic fibers and both muscarinic and nicotinic receptors. In decerebrate cats intravermal injection of the muscarinic agonist bethanechol increased the VSR gain. The cellular mechanisms underlying these gain changes were studied in anesthetized Sprague-Dawley rats by microiontophoresis. Application of bethanechol (10-60 nA, 300 s) increased the response of vermal P-cells to pulses of glutamate (22/33 cells) or GABA (23/25 cells). These effects, which were blocked by the muscarinic antagonist scopolamine, lasted up to 15-40 min and occurred regardless of whether bethanecol altered the basal firing rate of the cells. We propose that the increase of P-cell responses to both excitatory and inhibitory neurotransmitters following activation of muscarinic receptors enhances the amplitude of modulation of these neurons to animal tilt, thus increasing the gain of the VSR.

Depression of the vestibulospinal reflex adaptation by intravermal microinjection of GABA-A and GABA-B agonists in the cat.

In decerebrate cats the gain of the vestibulospinal reflex (VSR), elicited by sinusoidal roll tilt of the animal at 0.15 Hz, +/- 10 degrees, was tested every 10-15 min during and after a sustained (3 h) period of roll tilt of the head at the parameters indicated above, associated with synchronous roll tilt of the body at 0.15 Hz, +/- 12.5 degrees; this stimulus led to 2.5 degrees of neck rotation, which was thus out of phase with respect to head rotation. In this condition the gain of the VSR progressively increased during the first h of neck-vestibular stimulation, to reach a plateau level at the end of the third h of stimulation. This adaptive process was followed for at least 1 h after stimulation. Microinjection into the zone B of the cerebellar anterior vermis of the GABA-A agonist muscimol (0.25 microliter at 8 micrograms/microliter saline) producing only a slight or negligible depression of the VSR gain in non-adaptive conditions, prevented the occurrence of the adapted increase in gain of the VSR following a 3-h period of sustained head and neck rotation. In addition, intravermal injection of the GABA-A or the GABA-B agonist muscimol or baclofen, respectively, at the same dose indicated above supressed the adapted increase in gain occurring after a 3-h period of continuous neck-vestibular stimulation. The effective sites were located into the zone B of the cerebellar anterior vermis, from which the direct corticocerebellar projection to the lateral vestibular nucleus originates. In conclusion, the results seem to indicate that the adaptive increase in gain of the VSR which occurs in decerebrate cats depend upon plastic changes which affect the Purkinje cells of the cerebellar anterior vermis. These changes were in fact suppressed by GABAergic inhibition of these neurons. The demonstration that the effects of the GABA agonists occurred suddenly makes unlikely the hypothesis that the cerebellar anterior vermis represents either a relay for adaptive changes occurring before it (for instance in the inferior olive) or else the generator of error signals that elicit plasticity in target structures (as in the vestibular nuclei).