The cerebellar interposed nuclei (IN) are an essential part of circuits that control classically conditioned eyeblinks in the rabbit. The function of the IN is under the control of GABAergic projections from Purkinje cells of the cerebellar cortex. The exact involvement of cerebellar cortical input into the IN during eyeblink expression is not clear. While it is known that the application of gamma-aminobutyric acid-A (GABA(A)) agonists and antagonists affects the performance of classically conditioned eyeblinks, the effects of these drugs on IN neurons in vivo are not known. The purpose of the present study was to measure the effects of muscimol and picrotoxin on the expression of conditioned eyeblinks and the activity of IN cells simultaneously. Injections of muscimol abolished conditioned responses and either silenced or diminished the activity of IN cells. Two injections were administered in each picrotoxin experiment. The first injection of picrotoxin slightly modified the timing and amplitude of the eyeblink, produced mild tonic eyelid closure, increased tonic activity of IN cells, and reduced the amplitude of the neural responses. The second injection of picrotoxin abolished conditioned responses, further increased tonic eyelid closure, dramatically elevated the tonic activity of IN cells, and in most cases, abolished neuronal responses. These results demonstrate that both GABA(A)-mediated inactivation and tonic up-regulation of IN cells can interrupt the expression of conditioned eyeblinks and that this behavioral effect is accompanied by the suppression of the neuronal activity correlates of the conditioned stimulus and response.

No abstract

The cerebellar interposed nuclei are considered critical components of circuits controlling the classical conditioning of eyeblink responses in several mammalian species. The main purpose of the present experiments was to examine whether the interposed nuclei are also involved in the control of classically conditioned withdrawal responses in other skeletomuscular effector systems. To achieve this objective, a unique learning paradigm was developed to examine classically conditioned withdrawal responses in three effector systems (the eyelid, forelimb and hindlimb) in individual cats. Trained animals were injected with muscimol in the cerebellar interposed nuclei, and the effects on the three conditioned responses (CRs) were examined. Although the effects of muscimol were less dramatic than previously reported in the rabbit eyeblink preparation, the inactivation of the cerebellar nuclei affected the performance of CRs in all three effector systems. In additional experiments, animals were injected with muscimol at the sites affecting classically conditioned withdrawal responses to determine the effects of these injections on reaching and locomotion behaviors. These tests demonstrated that the same regions of the cerebellar interposed nuclei which control withdrawal reflexes are also involved in the control of limb flexion and precision placement of the paw during both locomotion and reaching tasks. The obtained data indicate that the interposed nuclei are involved in the control of ipsilateral action primitives and that inactivating the interposed nuclei affects several modes of action of these functional units.

No abstract

Classical conditioning of the eyeblink response is a form of motor learning that is controlled by the intermediate cerebellum and related brainstem structures. The inferior olive (IO) is commonly thought to provide the cerebellum with a "teaching" unconditioned stimulus (US) signal required for cerebellar learning. Testing this concept has been difficult because the IO, in addition to its putative learning function, also controls tonic activity in the cerebellum. Previously, it was reported that inactivation of AMPA/kainate receptors in the IO produces extinction of conditioned responses (CRs), suggesting that it blocks the transmission of US signals without perturbing the functional state of the cerebellum. However, the electrophysiological support for this critical finding was lacking, mostly because of methodological difficulties in maintaining stable recordings from the same set of single units throughout long drug injection sessions in awake rabbits. To address this critical issue, we used our microwire-based multiple single-unit recording method. The IO in trained rabbits was injected with the AMPA/kainate receptor blocker, 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide (NBQX), and its effects on CR expression and neuronal activity in the cerebellar interposed nuclei (IN) were examined. We found that NBQX abolished CR expression and that delayed drug effects were independent of the presentation of the conditioned stimulus and were therefore not related to extinction. In parallel to these behavioral effects, the spontaneous neuronal activity and CR-related neuronal responses in the IN were suppressed, suggesting cerebellar dysfunction. These findings indicate that testing the role of IO in learning requires methods that do not alter the functional state of the cerebellum.

Classical conditioning of the eyeblink reflex is a form of motor learning that is uniquely dependent on the cerebellum. The cerebellar learning hypothesis proposes that plasticity subserving eyeblink conditioning occurs in the cerebellum. The major evidence for this hypothesis originated from studies based on the telecommunications network metaphor of eyeblink circuits. These experiments inactivated parts of cerebellum-related networks during the acquisition and expression of classically conditioned eyeblinks in order to determine sites at which the plasticity occurred. However, recent evidence revealed that these manipulations could be explained by a network performance hypothesis which attributes learning deficits to a non-specific tonic dysfunction of eyeblink networks. Since eyeblink conditioning is mediated by a spontaneously active, recurrent neuronal network with strong tonic interactions, differentiating between the cerebellar learning hypothesis and the network performance hypothesis represents a major experimental challenge. A possible solution to this problem is offered by several promising new approaches that minimize the effects of experimental interventions on spontaneous neuronal activity. Results from these studies indicate that plastic changes underlying eyeblink conditioning are distributed across several cerebellar and extra-cerebellar regions. Specific input interactions that induce these plastic changes as well as their cellular mechanisms remain unresolved.

These experiments were designed to examine the effects of inactivating separately each of the major cerebellar nuclear regions in cats on the execution and retention of a previously learned, operantly conditioned volitional forelimb movement. The experiments test the postulates that the cerebellar nuclei, and particularly the interposed nuclei, contribute substantially to the spatial and temporal features of the interjoint coordination required to execute the task and that the engram necessary for the retention of this task is not located in any one of the cerebellar nuclei. All cats were trained to perform a task in which they were required to reach for and grasp a vertical bar at the sound of a tone and move the bar to a reward zone through a template consisting of two straight grooves in the shape of an inverted "L." After the task was learned, the effects of inactivating separately each nuclear region (the fastigial, interposed, and dentate nuclei) using muscimol microinjections were determined. Data were analyzed by quantifying several features of the movement's kinematics and by determining changes in the organization of the reaching component of the movement using an application of dimensionality analysis, an analysis that examines the correlation among the changes in joint angles and limb segment positions during the task. The retention of the previously learned task also was assessed after each injection. Injections of each nuclear region affected temporal and spatial features of the learned movement. However, the largest effects resulted from inactivating the interposed nuclei. These effects included an increased length of the reach trajectory, an accentuated deviation of the wrist trajectory from a straight line, cyclic movement of the distal extremity as the target was approached, a difficulty in grasping the bar, altered temporal features of the movement, and a highly characteristic change in the dimensionality measurements. The changes in dimensionality reflected a decreased correlation (linear interdependence) of the joint angular velocities coupled with an increased correlation among the linear velocities of markers located on the joints themselves. Related but less consistent changes in dimensionality resulted from fastigial injections. The motor sequence required to negotiate the template could be executed after the nuclear microinjections, indicating that retention of the motor sequence was not affected by the inactivation of any of the cerebellar nuclei. However, in two of the five animals, some decreases in performance were observed after dentate injection that were not characteristic of changes related to an effect on retention. These data suggest that the cerebellum plays an important role in regulating the consistent, stereotypic organization of complex goal-directed movements, including the temporal correlation among joint angle velocities. The data also indicate that the retention of the task is not dependent on any of the individual cerebellar nuclear regions. Consequently, these struct...

The purpose of these experiments was to examine the role of the human cerebellum in the acquisition and retention of conditioned reflexes. Normal human subjects and patients with cerebellar lesions were tested for their capacity to acquire, retain and express conditioned eyeblink responses. In acquisition tests, subjects were trained in a delay classical conditioning paradigm using a tone conditioned stimulus and a midline forehead tap as an unconditioned stimulus. While normal subjects developed anticipatory eyeblinks to the tone in one session, patients with cerebellar lesions failed to acquire conditioned responses in four consecutive training sessions. The conditioning deficit was bilateral even in patients with a unilateral cerebellar pathology. The same groups of subjects were tested for the presence of eyeblinks to a visual threat. In these experiments, both normal subjects and patients with cerebellar lesions exhibited a high level of responding when they saw an object approaching their face. These eyeblinks to the visual threat are probably naturally acquired conditioned responses because they extinguish in normal subjects if they are not reinforced by the unconditioned cutaneous stimulus. In addition, the stimulus of seeing an approaching object blocks the acquisition of classically conditioned eyeblinks to a new conditioned stimulus in normal subjects. These data imply that patients with cerebellar lesions who cannot acquire new classically conditioned responses are able to retain and express conditioned eyeblinks which were acquired before the onset of the pathology. Consequently, cerebellum-dependent neural substrates which are involved in learning new conditioned reflexes do not seem to be required for the storage of naturally learned conditioned responses.