Slowly-Conducting Pyramidal Tract Neurons in Macaque and Rat.

Anatomical studies report a large proportion of fine myelinated fibers in the primate pyramidal tract (PT), while very few PT neurons (PTNs) with slow conduction velocities (CV) (<~10 m/s) are reported electrophysiologically. This discrepancy might reflect recording bias toward fast PTNs or prevention of antidromic invasion by recurrent inhibition (RI) of slow PTNs from faster axons. We investigated these factors in recordings made with a polyprobe (32 closely-spaced contacts) from motor cortex of anesthetized rats (n = 2) and macaques (n = 3), concentrating our search on PTNs with long antidromic latencies (ADLs). We identified 21 rat PTNs with ADLs >2.6 ms and estimated CV 3-8 m/s, and 67 macaque PTNs (>3.9 ms, CV 6-12 m/s). Spikes of most slow PTNs were small and present on only some recording contacts, while spikes from simultaneously recorded fast-conducting PTNs were large and appeared on all contacts. Antidromic thresholds were similar for fast and slow PTNS, while spike duration was considerably longer in slow PTNs. Most slow PTNs showed no signs of failure to respond antidromically. A number of tests, including intracortical microinjection of bicuculline (GABAA antagonist), failed to provide any evidence that RI prevented antidromic invasion of slow PTNs. Our results suggest that recording bias is the main reason why previous studies were dominated by fast PTNs.

Functional Characterization of the Left Ventrolateral Premotor Cortex in Humans: A Direct Electrophysiological Approach.

In monkeys, motor outputs from premotor cortex (PM) involve cortico-cortical connections with primary motor cortex (M1). However, in humans, the functional organization of PM and its relationship with the corticospinal tract (CST) is still uncertain. This study was carried out in 21 patients undergoing intraoperative brain mapping prior to tumor resection. The left ventrolateral premotor cortex (vlPM-BA6) was identified preoperatively by functional magnetic resonance imaging, and then investigated intraoperatively using high frequency direct electrical stimulation (HF-DES) of the convexity of M1 and vlPM-BA6, with simultaneous recording of motor-evoked potentials (MEPs) from oro-facial, hand and arm muscles. The somatotopy, organization of evoked responses, latency of MEPs, and cortical excitability of vlPM-BA6 were compared with reference data from M1. vlPM-BA6 was found to be less excitable, with significantly longer MEP latencies than M1. In addition to the pure oro-facial and hand-arm muscle representation, a "transition oro-hand zone" was identified in vlPM-BA6. The longer latency of vlPM-BA6 MEPs suggests that human vlPM could act on spinal motoneurons either directly through more slowly conducting CST fibers or via less direct pathways through M1, brainstem, or spinal mechanisms. The results help in disclosing the very different roles of vlPM and M1 in motor control.

Do monkey F5 mirror neurons show changes in firing rate during repeated observation of natural actions?

Mirror neurons were first discovered in area F5 of macaque monkeys. In humans, noninvasive studies have demonstrated an increased blood oxygen level-dependent (BOLD) signal in homologous motor areas during action observation. One approach to demonstrating that this indicates the existence of mirror neurons in humans has been to employ functional (f)MRI adaptation to test whether the same population of neurons is active during both observation and execution conditions. Although a number of human studies have reported fMRI adaptation in these areas, a recent study has shown that macaque mirror neurons do not attenuate their firing rate with two repetitions. Here we investigated whether mirror neurons modulate their firing rate when monkeys observed the same repeated natural action multiple times. We recorded from 67 mirror neurons in area F5 of two macaque monkeys while they observed an experimenter perform a reach-to-grasp action on a small food reward using a precision grip. Although no changes were detectable for the first two repetitions, we show that both the firing rate and the latency at which mirror neurons discharged during observation were subtly modulated by the repetition of the observed action over 7-10 trials. Significant adaption was mostly found in the period immediately before the grasp was performed. We also found that the local field potential activity in F5 (beta-frequency range, 16-23 Hz), which is attenuated during action observation, also showed systematic changes with repeated observation. These LFP changes occurred well in advance of the mirror neuron adaptation. We conclude that macaque mirror neurons can show intra-modal adaptation, but whether this is related to fMRI adaptation of the BOLD signal requires further investigation.

Axon diameters and conduction velocities in the macaque pyramidal tract.

Small axons far outnumber larger fibers in the corticospinal tract, but the function of these small axons remains poorly understood. This is because they are difficult to identify, and therefore their physiology remains obscure. To assess the extent of the mismatch between anatomic and physiological measures, we compared conduction time and velocity in a large number of macaque corticospinal neurons with the distribution of axon diameters at the level of the medullary pyramid, using both light and electron microscopy. At the electron microscopic level, a total of 4,172 axons were sampled from 2 adult male macaque monkeys. We confirmed that there were virtually no unmyelinated fibers in the pyramidal tract. About 14% of pyramidal tract axons had a diameter smaller than 0.50 µm (including myelin sheath), most of these remaining undetected using light microscopy, and 52% were smaller than 1 µm. In the electrophysiological study, we determined the distribution of antidromic latencies of pyramidal tract neurons, recorded in primary motor cortex, ventral premotor cortex, and supplementary motor area and identified by pyramidal tract stimulation (799 pyramidal tract neurons, 7 adult awake macaques) or orthodromically from corticospinal axons recorded at the mid-cervical spinal level (192 axons, 5 adult anesthetized macaques). The distribution of antidromic and orthodromic latencies of corticospinal neurons was strongly biased toward those with large, fast-conducting axons. Axons smaller than 3 µm and with a conduction velocity below 18 m/s were grossly underrepresented in our electrophysiological recordings, and those below 1 µm (6 m/s) were probably not represented at all. The identity, location, and function of the majority of corticospinal neurons with small, slowly conducting axons remains unknown.

Gray and white matter changes associated with tool-use learning in macaque monkeys.

We used noninvasive MRI and voxel-based morphometry (VBM) to detect changes in brain structure in three adult Japanese macaques trained to use a rake to retrieve food rewards. Monkeys, who were naive to any previous tool use, were scanned repeatedly in a 4-T scanner over 6 weeks, comprising 2 weeks of habituation followed by 2 weeks of intensive daily training and a 2-week posttraining period. VBM analysis revealed significant increases in gray matter with rake performance across the three monkeys. The effects were most significant (P < 0.05 corrected for multiple comparisons across the whole brain) in the right superior temporal sulcus, right second somatosensory area, and right intraparietal sulcus, with less significant effects (P < 0.001 uncorrected) in these same regions of the left hemisphere. Bilateral increases were also observed in the white matter of the cerebellar hemisphere in lobule 5. In two of the monkeys who exhibited rapid learning of the rake task, gray matter volume in peak voxels increased by up to 17% during the intensive training period; the earliest changes were seen after 1 week of intensive training, and they generally peaked when performance on the task plateaued. In the third monkey, who was slower to learn the task, peak voxels showed no systematic changes. Thus, VBM can detect significant brain changes in individual trained monkeys exposed to tool-use training for the first time. This approach could open up a means of investigating the underlying neurobiology of motor learning and other higher brain functions in individual animals.

Population-averaged standard template brain atlas for the common marmoset (Callithrix jacchus).

Advanced magnetic resonance (MR) neuroimaging analysis techniques based on voxel-wise statistics, such as voxel-based morphometry (VBM) and functional MRI, are widely applied to cognitive brain research in both human subjects and in non-human primates. Recent developments in imaging have enabled the evaluation of smaller animal models with sufficient spatial resolution. The common marmoset (Callithrix jacchus), a small New World primate species, has been widely used in neuroscience research, to which voxel-wise statistics could be extended with a species-specific brain template. Here, we report, for the first time, a tissue-segmented, population-averaged standard template of the common marmoset brain. This template was created by using anatomical T(1)-weighted images from 22 adult marmosets with a high-resolution isotropic voxel size of (0.2 mm)(3) at 7-Tesla and DARTEL algorithm in SPM8. Whole brain templates are available at International Neuroinformatics Japan Node website, http://brainatlas.brain.riken.jp/marmoset/.

Creating a population-averaged standard brain template for Japanese macaques (M. fuscata).

A number of modern digital anatomy techniques, based on structural MR brain images, have recently become applicable to the non-human primate brain. Such voxel-based quantitative techniques require a species-specific standardized brain template. Here we present a brain template for the Japanese macaque (Macaca fuscata). The template was designed to be used as a tool for spatially normalising Japanese macaque brains into a standard space. Although this species of macaque monkey is widely used in neuroscience research, including studies of higher cognitive brain functions, no standard MRI template of its brain is presently available. The template presented here is based on T1/T2* weighted, high-resolution 4T MR images obtained from 16 male adult Japanese macaque monkeys. T1/T2* images were used to correct the signal inequalities resulting from the use of a surface coil. Based on these images, population-averaged probability maps were created for grey matter, white matter and cerebrospinal fluid. The new template presented here should facilitate future brain research using the Japanese macaque monkey. Whole brain templates are available at http://brainatlas.brain.riken.jp/jm/modules/xoonips/listitem.php?index_id=9.

Motor control: forcing neurons to change.

New research shows that the properties of cells in motor cortex change during learning of new tasks, shedding new light on the neural basis of motor adaptation.

Neural changes in the primate brain correlated with the evolution of complex motor skills.

Complex motor skills of eventual benefit can be learned after considerable trial and error. What do structural brain changes that accompany such effortful long-term learning tell us about the mechanisms for developing innovative behavior? Using MRI, we monitored brain structure before, during and after four marmosets learnt to use a rake, over a long period of 10-13 months. Throughout learning, improvements in dexterity and visuo-motor co-ordination correlated with increased volume in the lateral extrastriate cortex. During late learning, when the most complex behavior was maintained by sustained motivation to acquire the skill, the volume of the nucleus accumbens increased. These findings reflect the motivational state required to learn, and show accelerated function in higher visual cortex that is consistent with neurocognitive divergence across a spectrum of primate species.

Macaque ventral premotor cortex exerts powerful facilitation of motor cortex outputs to upper limb motoneurons.

The ventral premotor area (F5) is part of the cortical circuit controlling visuomotor grasp. F5 could influence hand motor function through at least two pathways: corticospinal projections and corticocortical projections to primary motor cortex (M1). We found that stimulation of macaque F5, which by itself evoked little or no detectable corticospinal output, could produce a robust modulation of motor outputs from M1. Arrays of fine microwires were implanted in F5 and M1. During terminal experiments under chloralose anesthesia, single stimuli delivered to M1 electrodes evoked direct (D) and indirect (I1,I2, and I3) corticospinal volleys. In contrast, single F5 shocks were ineffective; double shocks (3 msec separation) evoked small I waves but no D wave. However, when the test (T) M1 shock was conditioned (C) by single or double F5 shocks, there was strong facilitation of I2 and I3 waves from M1, with C-T intervals of <1 msec. Intracellular recordings from 79 arm and hand motoneurons (MNs) revealed no postsynaptic effects from single F5 shocks. In contrast, these stimuli produced a robust facilitation of I2 and I3 EPSPs evoked from M1 (60% of MNs); this was particularly marked in hand muscle MNs (92%). Muscimol injection in M1 reduced I waves from F5 and abolished the F5-induced facilitation of late I waves from M1, and of EPSPs associated with them. Thus, some motor effects evoked from F5 may be mediated by corticocortical inputs to M1 impinging on interneurons generating late corticospinal I waves. Similar mechanisms may allow F5 to modulate grasp-related outputs from M1.

Human cortical muscle coherence is directly related to specific motor parameters.

Cortical oscillations have been the target of many recent investigations, because it has been proposed that they could function to solve the "binding" problem. In the motor cortex, oscillatory activity has been reported at a variety of frequencies between approximately 4 and approximately 60 Hz. Previous research has shown that 15-30 Hz oscillatory activity in the primary motor cortex is coherent or phase locked to activity in contralateral hand and forearm muscles during isometric contractions. However, the function of this oscillatory activity remains unclear. Is it simply an epiphenomenon or is it related to specific motor parameters? In this study, we investigated task-dependent modulation in coherence between motor cortex and hand muscles during precision grip tasks. Twelve right-handed subjects used index finger and thumb to grip two levers that were under robotic control. Each lever was fitted with a sensitive force gauge. Subjects received visual feedback of lever force levels and were instructed to keep them within target boxes throughout each trial. Surface EMGs were recorded from four hand and forearm muscles, and magnetoencephalography (MEG) was recorded using a 306 channel neuromagnetometer. All subjects showed significant levels of coherence (0.086-0.599) between MEG and muscle in the 15-30 Hz range. Coherence was significantly smaller when the task was performed under an isometric condition (levers fixed) compared with a compliant condition in which subjects moved the levers against a spring-like load. Furthermore, there was a positive, significant relationship between the level of coherence and the degree of lever compliance. These results argue in favor of coherence between cortex and muscle being related to specific parameters of hand motor function.

The importance of the cortico-motoneuronal system for control of grasp.

Our recent work has revealed new evidence of the importance of direct cortico-motoneuronal (CM) connections for voluntary control of the hand. Most of these connections are derived from corticospinal neurons located in the M1 hand area, although there are some much smaller contributions from other secondary motor areas, such as the supplementary motor area (SMA). Intracellular recordings show that 75% of upper limb motoneurons in the chloralose-anaesthetized macaque monkey receive a monosynaptic projection from the corticospinal tract; evidence for non-monosynaptic, propriospinal excitatory influences from the corticospinal tract was conspicuously lacking in these anaesthetized preparations. Moreover, in the conscious monkey, hand and arm muscle motor unit responses to corticospinal tract input are dominated by single, brief peaks compatible with monosynaptic excitation. CM excitatory post-synaptic potentials, recorded from a comparable sample of hand and arm motoneurons in anaesthetized macaque and squirrel monkeys, were found to be larger and faster rising in the macaque, which is by far the more dexterous of the two species. CM cells facilitating a given muscle in the conscious macaque are distributed over a wide region of M1 cortex, and each contributes a particular pattern of discharge during a skilled task. In addition to their direct effects on target muscles there may be weaker but potentially important effects that derive from the synchronous binding of assemblies of output neurons. Synchronous oscillations between these neurons are particularly prevalent during steady grip, but disappear during digit movement.

Cerebral activation during the exertion of sustained static force in man.

The aim of our study was to determine alterations of cerebral activity during prolonged static force exertion. Regional cerebral blood flow (rCBF) was measured using H2(15)O positron emission tomography (PET) while six male normal subjects pressed a morse-key with their right index finger with a constant force of 20% of their maximal voluntary contraction (MVC) for different periods of time (1.5-4.5 min). Exertion of static force led to activation which was at least as extensive as that during exertion of repetitive dynamic force pulses. Despite a considerable sense of fatigue and increased effort at the end of a 4.5 min key press, no compensatory changes of activity were detected in motor or sensory related structures. The right dorsolateral prefrontal cortex demonstrated a significant correlation between rCBF and duration of key-press, possibly reflecting processes over-riding fatigue. Prominent basal ganglia activation was demonstrated in this static force task, but not in a previous force task involving repetitive dynamic force pulses. This suggests that sustained exertion of a static force is an active process modulated, at least in part, by the basal ganglia.

Comparison of regional cerebral blood flow with transcranial magnetic stimulation at different forces.

This study's objective was to investigate regional cerebral blood flow (rCBF) within the primary motor cortex (M1) and to compare it with thresholds of transcranial magnetic stimulation (TMS) and electromyographic recordings during exertion of different force levels with the right index finger. Quantitative electromyographic recordings, TMS, and positron emission tomography scans were performed while five and six volunteers, respectively, pressed a Morse key repetitively or with constant force with the right hand at five different force levels: 5, 10, 20, 40, and 60% of the individual's maximum voluntary contraction (MVC). Although at 5% MVC muscle activity was restricted to the first dorsal interosseus muscle, superficial finger flexors, and extensors, there was progressive involvement of proximal muscles during finger flexion with increasing force. rCBF increased logarithmically in the contralateral M1 with increasing force. In ipsilateral M1, rCBF decreased at 5% MVC and then increased logarithmically at higher force levels. TMS thresholds in the contralateral hemisphere declined logarithmically to reach a plateau at high force levels. The threshold in the ipsilateral hemisphere decreased slightly at high force levels. The logarithmic increase of rCBF and decrease of TMS thresholds in the contralateral hemisphere suggest related underlying physiological phenomena; increased cortical synaptic activity and increased excitability. It suggested that the pronounced ipsilateral rCBF alterations reflect transcallosal inhibition and are more prominent during repetitive movements (as used in the positron emission tomography study) than during the generation of a constant force (as exerted during TMS).

Non-linear summation of responses in averages of rectified EMG.

Studies on the motor system commonly use averages of rectified electromyogram (EMG) to measure muscular response. The assumption is usually made that this is a linear measure of response magnitude. It is shown here using a theoretical and experimental model that averages of rectified EMG can lead to non-linearities, which are of particular importance when considering the summation of two independent responses. When responses had a highly stereotyped waveform from trial to trial, so that averages of unrectified EMG revealed a deflection from zero, in measurements from averages of rectified EMG the size of a response to two stimuli delivered together was greater than the linear sum of the responses to each stimulus delivered alone. This deviation from linearity was greatest when two responses which were small relative to the ongoing background EMG activity were combined. The total response was then as much as twice as large as the linear sum of the individual component responses. Conversely, under conditions where responses were highly variable from trial to trial, resulting in no consistent deflection being seen in an average of unrectified EMG, the summed responses in rectified and averaged EMG were smaller than expected if summation were linear. This effect was most pronounced when both component responses were relatively large; the combined response tended asymptotically to approximately 70% of the linear sum of the component responses. A practical method is presented which allows prediction of the size of a response to two stimuli given together, when measured from averages of rectified EMG, on the assumption that they act independently.

Antidromic excitation of motoneurons by intramuscular electrical stimulation.

The terminal branches of motoneurons can be excited by electrical current pulses delivered via needles inserted intramuscularly. Observations are reported which indicate that tungsten semi-microelectrodes suitably positioned in a muscle belly can excite antidromically a large proportion of the alpha-motoneurons of that muscle, at stimulus strengths weak enough to avoid spread of excitation to motor units of adjacent muscles. Some gamma-motoneurons and afferent axons can also be excited by intramuscular stimulation. This technique of antidromic excitation will find application in electrophysiological studies on motoneurons of small muscles, such as intrinsic hand and foot musculature, where the individual muscle nerves are too fragile to be dissected and mounted on stimulating electrodes in the conventional manner.

Multiple single unit recording in the cortex of monkeys using independently moveable microelectrodes.

Simultaneous recording from multiple single neurones presents many technical difficulties. However, obtaining such data has many advantages, which make it highly worthwhile to overcome the technical problems. This report describes methods which we have developed to permit recordings in awake behaving monkeys using the 'Eckhorn' 16 electrode microdrive. Structural magnetic resonance images are collected to guide electrode placement. Head fixation is achieved using a specially designed headpiece, modified for the multiple electrode approach, and access to the cortex is provided via a novel recording chamber. Growth of scar tissue over the exposed dura mater is reduced using an anti-mitotic compound. Control of the microdrive is achieved by a computerised system which permits several experimenters to move different electrodes simultaneously, considerably reducing the load on an individual operator. Neurones are identified as pyramidal tract neurones by antidromic stimulation through chronically implanted electrodes; stimulus control is integrated into the computerised system. Finally, analysis of multiple single unit recordings requires accurate methods to correct for non-stationarity in unit firing. A novel technique for such correction is discussed.

The involvement of monkey premotor cortex neurones in preparation of visually cued arm movements.

Some neurones in macaque postarcuate premotor area modulate their firing frequency in relation to motor tasks which require visual information. We previously reported that a large proportion of these neurones modulate during execution of a detour reaching task in which the movement phase was separated in time from the phase in which the monkey received a visual cue for the movement required to retrieve a food reward. A large proportion of task-related neurones (75%) modulated during this 'visual' phase, in which no task-related movements were made. This modulation was related to the position of the food reward, which served as the visual cue. Most of these neurones were located in cortical area 6, close to the arcuate curvature and its spur, but also more caudally in area 4 and rostrally in area 8. In the present chronic recording experiments in monkeys, several variations of the original task were used in order to test whether the 'visual'-related neuronal modulation could be involved in preparation of the upcoming movement. This modulation is unlikely to be related to any eye or arm movements occurring during the visual phase or to changes in environmental illumination. Neither can it be related to the presence of the visual cue in a particular part of the visual field, since the pattern of neuronal modulation was similar when a cue with a fixed position was used. This modulation was, however, contingent upon the occurrence of food retrieval during the subsequent 'movement phase', since it was abolished or diminished during presentation of a 'food-reward' which the monkey did not retrieve. For several neurones, modulation pattern during the visual phase depended on whether the food reward was to be retrieved with a gross hand movement or with relatively independent finger movements. It is likely, therefore, that neurones in the postarcuate premotor cortex are involved in preparation for arm movements with the help of visual cues. The results are discussed in view of corticocortical pathways which might be involved in transmission of visual information from visual areas through parietal association areas and premotor cortex to the primary motor cortex.

Comparison of direct and indirect measurements of the central motor conduction time in the monkey.

OBJECTIVE: The goal of the present study was to compare the indirect estimate of the central motor conduction time (CMCT) with direct measurement of the corticospinal conduction time between the motor cortex and cervical enlargement in macaque monkeys. METHODS: Responses to transcranial magnetic stimulation of the motor cortex were recorded from intrinsic hand muscles in adult macaque monkeys. The CMCT was calculated by subtracting the peripheral conduction time, measured with the F-wave method, from the latency of the motor evoked potentials (MEPs). In two monkeys, the actual conduction time between the motor cortex and cervical enlargement was measured directly by different invasive techniques. RESULTS: We found that the indirect calculation of CMCT overestimates the corticospinal conduction time to a significant extent. CONCLUSIONS: One possible source of error is an underestimate of the MEP peripheral conduction time. A collision test confirmed this hypothesis and showed that only a marginal proportion of the motoneurones that respond to a weak corticospinal input also participate in the F-wave. A more accurate estimate of the CMCT could be obtained by using the longest F-wave latency, rather than the shortest, to calculate the peripheral conduction time.