Timothy J. Ebner

Sustained release of nerve growth factor from biodegradable polymer microspheres.

Although grafted adrenal medullary tissue to the striatum has been used both experimentally and clinically in parkinsonism, there is a definite need to augment long-term survival. Infusion of nerve growth factor (NGF) or implantation of NGF-rich tissue into the area of the graft prolongs survival and induces differentiation into neural-like cells. To provide for prolonged, site-specific delivery of this growth factor to the grafted tissue in a convenient manner, we fabricated biodegradable polymer microspheres of poly(L-lactide)co-glycolide (70:30) containing NGF. Biologically active NGF was released from the microspheres, as assayed by neurite outgrowth in a dorsal root ganglion tissue culture system. Anti-NGF could block this outgrowth. An enzyme-linked immunosorbent assay detected NGF still being released in vitro for longer than 5 weeks. In vivo immunohistochemical studies showed release over a 4.5-week period. This technique should prove useful for incorporating NGF and other growth factors into polymers and delivering proteins and other macromolecules intracerebrally over a prolonged time period. These growth factor-containing polymer microspheres can be used in work aimed at prolonging graft survival, treating experimental Alzheimers disease, and augmenting peripheral nerve regeneration.

Climbing fiber afferent modulation during a visually guided, multi-joint arm movement in the monkey

During a visually guided, multi-joint voluntary arm movement Purkinje cell simple and complex spike activity was recorded from the ipsilateral hemisphere and intermediate zone of the cerebellum in the rhesus monkey. The task consisted of moving a manipulandum over a horizontal video screen. Manipulandum (hand position) was represented by a cursor on the screen, the animal required to place the manipulandum within displayed start and target boxes. Purkinje cell complex spike discharge was examined using two paradigms. In the first the animal moved the manipulandum from a start box to a target box. In the second the animal was required to modify an ongoing movement and place the cursor within a repositioned target box. A majority of the cells (44/74) exhibited a statistically significant increase in the probability of complex spike discharge at various times during the movement. The increase was observed when the movement trajectory was redirected (36/44) and/or during the initial portion of the movement (27/44). These results suggest the climbing fiber afferent system is routinely involved in the execution of multi-joint movements especially when the movement is redirected. Possibilities include that climbing fiber afferent input is required when the motor state changes and/or during errors in motor performance.

Functional magnetic resonance imaging of cerebellar activation during the learning of a visuomotor dissociation task

We have used functional magnetic resonance imaging (fMRI) to study the changes in cerebellar activation that occur during the acquisition of motor skill in human subjects presented with a new task. The standard paradigm consisted of a center‐out movement in which subjects used a joystick to superimposed a cursor onto viusual targets. Two variations of this paradigm were introduced: (1) a learning paradigm, where the relationship between movement of the joystick and cursor was reversed, requiring the learning of a visuomotor transformation to optimize performance and (2) a random paradigm, where the joystick/cursor relationship was changed randomly for each trial. Activation in the cerebellum was highest during the random paradigm and during the early stages of the learning paradigm. In the early stages of learning and during the random paradigm performance was poor with a decrease in the number of completed movements, and an increase in the time and length of movements. With repeated practice at the learning paradigm performance improbed and reached the same level of proficiency as in the standard task. Commensurate with the improbement in performance was a decrease in cerebellar activation, that is, activation in the cerebellum changed in a parallel, but inverse relationship with performance. Linear regression analysis demonstarated that the inverse correlation between cerebellar activation and motor performance was significant. Repeated practice at the random paradigm did not produce improvements in performance and cerebellar activity remained high. The data support the hypothesis that the cerebellum is strongly activated when motor performance is inaccurate, consistent with a role for the cerebellum in the detection of, and correction for visuomotor errors.

Functional magnetic resonance imaging of motor, sensory, and posterior parietal cortical areas during performance of sequential typing movements

Abstract We investigated the activation of sensory and motor areas involved in the production of typing movements using functional magnetic resonance imaging (fMRI). Eleven experienced typists performed tasks, in which the spatial and temporal requirements as well as the number of digits involved were varied. These included a simple uni-digit repetitive task, a uni-digit sequential task, a dual-digit sequential task, a multi-digit sequential task, and typing text from memory. We found that the production of simple repetitive keypresses with the index finger primarily involved the activation of contralateral primary motor cortex (M1), although a small activation of the supplementary motor area (SMA) and other regions was sometimes observed as well. The sequencing of keypresses involved bilateral M1 and a stronger activation of the SMA and to a lesser extent the premotor area, cingulate gyrus, caudate, and lentiform nuclei. However, the activation of these areas did not exclusively depend on the complexity of the movements, since they were often activated during more simple movements, such as alternating two keypresses repeatedly. Somatosensory and parietal regions were also found to be activated during typing sequences. The activation of parietal areas did not exclusively depend on the spatial requirements of the task, since similar activation was observed during movements within intra-personal space (finger-thumb opposition) and may instead be related to the temporal requirements of the task. Our findings suggest that the assembly of well-learned, goal-directed finger movement sequences involves the SMA and other secondary motor areas as well as somatosensory and parietal areas.

The changes in Purkinje cell simple spike activity following spontaneous climbing fiber inputs

The purpose of these experiments was to systematically examine the characteristics of the excitability change occurring after the inactivation period evoked by the climbing fiber input to Purkinje cells. Ninety-eight Purkinje cells were isolated extracellularly in unanesthetized decerebrate cats. Simple spikes and complex spikes were discriminated separately. Post-stimulus time histograms were constructed from 100 consecutive trials triggered by the occurrence of spontaneous complex spikes. Seventeen Purkinje cells exhibited a reduction of simple spike discharge rate following the inactivation period. However, 14 cells showed no change in simple spike activity, and in 67 cells the discharge rate increased. These changes in excitability following a spontaneous complex spike were independent of the tonic simple spike activity of the Purkinje cell. Single traces of spike train data from Purkinje cells showed that the change in discharge rate was variable, some complex spikes being followed by an increase and others by a decrease in activity. The basis for these observations and the differences between these data and those from studies in which the climbing fiber input was evoked by electrical olivary stimulation are discussed.

Responses of interposed and dentate neurons to perturbations of the locomotor cycle

SummaryThis study examined the relationship of antidromically identified neurons in the dentate and interposed nuclei to perturbed and unperturbed locomotion in the pre-collicular, mid-mamillary, decerebrate cat. During treadmill locomotion two methods were used to perturb the step cycle. In the first, the treadmill was braked in different phases of the step cycle, the “treadmill” perturbation. In the second, the motion of the ipsilateral forelimb was interrupted by a rod placed transiently in the limbs path, the “single limb” perturbation. Most interposed cells were modulated during locomotion, their discharge being highly correlated with the EMG of the ipsilateral biceps or triceps. When the locomotion was perturbed, the modulation ceased for the duration of the perturbation. A few interposed cells displayed activity patterns unrelated to the EMG but were responsive to perturbations of a single limb. These responses may be explained by the putative activation of peripheral afferents produced by the perturbation. Most dentate cells were not modulated during unperturbed locomotion but did respond to features of the treadmill perturbation. Usually the response was coupled to the resumption of treadmill motion. A minority of dentate neurons was modulated slightly during unperturbed locomotion. Their modulation was less dramatic than that of interposed cells and was only weakly related to limb movement or EMG activity. Like the interposed neurons, these dentate cells responded to the treadmill perturbation with a cessation of modulation. All dentate cells were unresponsive to single limb perturbations. In a preparation lacking cerebral cortical input, the findings show that neurons of the interposed and dentate nuclei are modulated differently during perturbed and unperturbed treadmill walking in the decerebrate cat. The activity of interposed neurons is related to specific features of EMG activity recorded from muscles in the ipsilateral forelimb. Although some dentate cells were weakly modulated during unperturbed locomotion, the majority of these neurons responded most dramatically to the occurrence of a perturbation which completely stopped the walking behavior.

Position, Direction of Movement, and Speed Tuning of Cerebellar Purkinje Cells during Circular Manual Tracking in Monkey

The cerebellum plays an essential role in pursuit tracking with the eye and with the hand. During smooth pursuit eye movements, both tracking position and velocity are signaled by Purkinje cells. Purkinje cell simple spike discharge is also modulated by direction and speed during linear manual tracking. This study evaluated how all three parameters, position, movement direction, and speed, are signaled in the simple spike discharge of Purkinje cells during circular manual tracking. Three rhesus monkeys intercepted and then tracked a target moving in a circle in both counterclockwise and clockwise directions across a range of constant target speeds. Two sets of analyses of the simple spike firing of 97 Purkinje cells examined the effects of position, movement direction, and speed. The first approach was the incremental improvement of regression models, initially modeling a pure position dependence, then incorporating movement direction, and finally incorporating speed dependence. The second was a model-independent approach, without any explicit assumptions about the character of the directional tuning or speed effects. Both analyses revealed the same three results: (1) Purkinje cell discharge is spatially tuned, to both the position and direction of movement, and (2) this spatial tuning is not altered by the speed, except (3) the speed scales the average firing and/or depth of modulation. The results suggest that the population of Purkinje cells forms a representation of the entire position-direction space of arm movements, and that the speed modulates the scale of that representation. This speed scaling provides insights into the cerebellar processing of movement-related timing.

What Do Complex Spikes Signal about Limb Movements

Abstract: Deciphering the information or signals carried by the complex spike discharge of Purkinje cells has proven to be problematic, primarily because of low frequency discharge and lack of adequate analytical techniques. This problem is particularly acute for studies of limb movements. To this end the relationship of cerebellar Purkinje cell complex spike discharge to direction and speed were studied in a manual‐tracking task. Two monkeys were trained to pursue track targets moving in one of eight directions and at one of four speeds. An analysis based on Poisson regression modeling fitted the complex spike counts during single movement trials to target direction and/or speed. Using single trial data, the Poisson modeling demonstrated that the complex spike discharge for a majority of the Purkinje cells was significantly fit to tracking direction and speed. A second analysis based on the directional distribution of position and speed errors and a Poisson regression model of complex spike discharge to tracking position and speed errors found little relationship to movement error. Comparison of the preferred direction of the complex spike discharge with that of the simple spike activity revealed a reciprocal relationship for many cells. Thus, the complex spike discharge signals both tracking direction and speed but not movement errors. Furthermore, treating complex spike counts as a Poisson process provides a powerful tool for analyzing these events in single trials, without the need for extensive averaging.

Changes in the responses of cerebellar nuclear neurons associated with the climbing fiber response of Purkinje cells.

Previous studies demonstrated that climbing fiber activity produces a short-term increase in the responsiveness of Purkinje cells to mossy fiber inputs. This led to the hypothesis that there are concomitant alterations in the discharge of cerebellar nuclear neurons. This series of experiments was initiated to test this hypothesis in simultaneously recorded Purkinje cell-nuclear cell pairs in related regions of the cerebellar cortex and nuclei. In decerebrate cats 50 pairs of Purkinje cells and nuclear neurons were identified and simultaneously recorded during spontaneous activity and during peripheral inputs. Auto-correlograms of nuclear cell activity and cross-correlograms of the simple spike and nuclear cell activity triggered on the occurrence of spontaneous complex spikes demonstrated little correlation between these events and the discharge of nuclear neurons. To examine the effect of evoked climbing fiber inputs on the Purkinje cell simple spike and nuclear cell responses, square wave mechanical stimuli which modulated the discharge of both cells of a pair were applied to the forepaw. A separation technique was used to construct one histogram illustrating the responses of the nuclear neuron and Purkinje cell in trials in which the peripheral stimulus evoked a climbing fiber input to the Purkinje cell and another histogram showing their responses in trials in which no climbing fiber input was activated. Using this separation technique it was shown that the amplitude of most Purkinje cell responses increased by 120-1200% in trials in which climbing fiber inputs were activated. The response amplitude of 68% of the nuclear cells was modified for these pairs. Most changes in nuclear cell responses were increases ranging from 120-220%. These changes were felt to reflect the action of many Purkinje cells converging on the isolated nuclear neuron. The modulation of the nuclear neuron was not due only to the effect of the related Purkinje cell, since the gain change of the Purkinje cell and nuclear cell of each pair was not correlated (r = 0.01). The discussion of these findings emphasizes that the increased responses of the nuclear cell are most likely produced by the intracortical action of the climbing fiber system on the responsiveness of Purkinje cells to mossy fiber inputs. Climbing fiber collateral input to nuclear neurons also may contribute to the changes in the nuclear cell responses observed in these experiments.

8–12 Hz rhythmic oscillations in human motor cortex during two-dimensional arm movements: evidence for representation of kinematic parameters ☆

Direct cortical recordings were taken from 12 patients with implanted subdural electrode arrays during performance of a 2-dimensional, multi-joint, visually guided arm movement task. Task-related changes in the amplitude of the motor cortex 8-12 Hz surface local field oscillations were evaluated for the encoding of direction and amplitude of movement in the 6 patients in whom no epileptogenic or ECoG background abnormalities were detected over the motor-sensory cortical areas under the recording electrode array. The topography, time of onset and duration of these responses were evaluated in the context of motor cortex somatotopy, as defined by cortical stimulation delivered through the electrode array. Multi-joint arm movements were accompanied by a decrease in the power of the 8-12 Hz frequency components of the ECoG signal. These power changes were spatially distributed over the upper extremity, motor-sensory representation. Movement amplitude influenced the magnitude, duration, and extent of the spatial distribution of ECoG power changes in the 8-12 Hz band. These effects occurred predominantly over cortical areas corresponding to the upper extremity motor-sensory representations. Direction of movement had a weaker influence on the 8-12 Hz frequency components of the ECoG over the upper extremity motor-sensory representations, but influenced the patterns of 8-12 Hz ECoG response on adjacent cortical regions. These results show that the amplitude of surface electrical oscillations generated over the rolandic cortex are correlated with the kinematics of multi-joint arm movements. These changes in the ECoG signal appear to reflect shifts in the functional state of neuronal ensembles involved in the initiation and execution of motor tasks.