a small laminotomy. Each electrode consisted of a 50 micron shaft of. 80-20% Pt-Ir alloy welded to a twisted pair of very flexible, 25 micron diameter, gold wires ...
ACTIVITY PATTERNS OF IDENTIFIED ALPHA MOTONEURONS TO CAT ANTERIOR THIGH MUSCLES DURING NORMAL WALKING AND FLEXOR REFLEXES
G. E. Loeb*, J. A. Hoffer, N. Sugano, W. B. Marks M. J. O'Donovan, and C. A. Pratt Laboratory of Neural Control National Institutes of Health Bethesda, MD INTRODUCTION The organization of the motor apparatus into anatomically and functionally defined pools of regularly recruited motor units derives from some of the earliest and most enduring observations of neurophysiology. In recent years, there has been much productive research concentrated on discovering the anatomical bases of this organization in the spinal cord circuitry and the properties of the final common pathway, the alpha motoneurons themselves (for review, see Burke, 1981a). However, almost all of the direct evidence for the function of this system has been derived from experiments on reduced or anesthetized animals and on human subjects performing highly constrained and artificial motor tasks. Over the past few years, we have been able to obtain recordings in intact, normally walking cats from single ventral root axons projecting to identified hindlimb muscles. This paper presents a brief summay of the most salient features noted in these studies (full reports are in preparation; abstracts of these data have been presented elsewhere). In general, the results have confirmed the broad outlines of current motor recruitment theories, while emphasizing new or overlooked features such as the importance of rate modulation and the complexities of multifunctional muscles. METHODS Seventeen adult cats were implanted chronically with a variety of recording electrodes and transducers, using general anesthesia and aseptic surgical techniques. Data from a total of 164 axons in the fifth lumbar ventral roots (L5 VR) were recorded during normal motor behavior at 1-10 weeks postoperatively, using a 40 pin percutaneous connector and multichannel analog and video recording equipment to provide correlary kinesiological information from the hindlimb.
*Correspondence: G. E. Loeb, National Institutes of Health, Bldg. 36, Rm. 5A29, 9000 Rockville Pike, Bethesda, MD 20892
Unit Recording and Identification Up to twelve floating microelectrodes were implanted in L5 VR via a small laminotomy. Each electrode consisted of a 50 micron shaft of 80-20% Pt-Ir alloy welded to a twisted pair of very flexible, 25 micron diameter, gold wires, in turn welded to stranded stainless steel cable for percutaneous exit, with the entire assembly overcoated with 15 microns of Parylene-C polymer (Loeb et al., 1977), and strain-relieved by passing through a short piece of silicone rubber tubing affixed to the dorsal spines. Once implanted, each electrode was tested daily for impedance (typically 80-220 kilohms at 1 kHz) and the presence of discriminable single unit action potentials during walking on a treadmill. Spike-triggered signal averaging was used successfully on 43 units to identify both the efferent conduction velocity and the muscle of destination. The femoral nerve (to which about half of the L5 segment projects) was chronically instrumented with a nerve cuff electrode that provided two adjacent tripolar recording sites for averaging the efferent action potentials from the triggering unit (Hoffer et al., 1981). The complete motor projections of the femoral nerve distal to the cuff include the monarticular vasti (lateralis-VL, medialis-VM, and intermedius-VI) and biarticular rectus femoris-RF and sartorius (pars medialis-SA-m and anterior-SA-a), all of which were chronically implanted with bipolar recording electrodes. Spike-triggered averaging of all of these EMG records usually revealed one predominant record, by which we identified the muscle innervated by the triggering unit. Neurokinesiological Data Processing The locomotor cycle was recorded by a high resolution video system with a time code to permit correlation with the analog signals recorded on 18-track FM tape (DC-10 kHz). All unit identification and analysis was performed off-line on the same recorded sequences, using conventional spike-discrimination equipment (Bak and Schmidt, 1977) and analog rectification and integration of EMG activity (Bak and Loeb, 1979) prior to digitization and computer analysis. For each motor unit, covariances were calculated between the instantaneous frequencygram (inverse of interspike interval) and the complete set of EMG signals (digitally smoothed with about 30 msec time-constant). The limb was also implanted with length gauges (saline-filled lengths of elastic silicone rubber tubing with electrodes in each end; Loeb et al., 1980) across the vasti muscle group (pure knee extensors) and RF-SA-a group (knee extensors plus hip flexors) These records plus their electronic time derivatives (velocity) are shown in the figures, along with the output of an implanted tendon strain gauge on the patellar ligament (Walmsley et al., 1978), where all muscles except SA-m insert.
A nerve cuff electrode on the saphenous nerve was electronicaUy stimulated with single current pulses (0.1 msec duration, 1.1-10 times threshold for group I fibers) to provide segmental cutaneous perturbations at random points in the step cycles. The prestimulus controls and flexor reflex responses of the various muscles and 11 motor axons were ordered into rasters on the basis of the phase in the step cycle at which each stimulus occurred (Abraham et al., 1985). FINDINGS Figures 1 and 2 show typical sets of tracings of unit frequencygrams plus rectified EMG from various muscles, including that innervated by the
Stance T a s k Group SA Motoneuron
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Fig. 1. Activity of a ventral root axon projecting to anterior sartorius muscle, during four normal walking step cycles (heavy bars denote stance phase). Traces from top down: frequency of unit firing, window discriminator output, velocity of muscle (stretch upward), length of muscle, rectified EMG integrated into 2 msec bins from anterior sartorius, medial sartorius, rectus femoris, and vastus medialis muscles, force output at patellar ligament strain gauge, and treadmill speed. Note unit activity only during stance phase EMG activity of parent muscle SA-a.
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unit, and length and force records during unperturbed treadmill walking. All of the motor units tended to be recruited in a single burst of activity each step cycle, beginning firing at about 8-15 pps whenever the amplitude of the parent muscle EMG exceeded a threshold that was typical for each unit and relatively independent of walking speed. The unit firing rate was always well-modulated in close correlation with parent muscle EMG, and could approach 50 pps even for first-recruited, presumably slow-twitch motoneurons. Units recruited at less than 50% of the peak EMG level tended to have axon conduction velocities of 50-90 m/s (8 of 10 units) whereas those recruited above 50% had axon conduction velocities of 90-120 m/s (14 of 18 units). Initial doublets (interspike intervals of less than 20 msec at the beginning of recruitment) were seen in only four out of 51 units. The two motoneurons shown in the figures are of particular interest because they have typical single activity bursts per step cycle despite the fact that their parent muscle, SA-a, had two EMG bursts per step cycle. Unit L3A5 in Figure 1 was active only during and in proportion to the stance phase muscle activity, whereas unit L2A42 in Figure 2 was active only during and in proportion to the swing phase muscl-e activity. Even when the EMG in the opposite phase exceeded levels usually associated with unit recruitment, all 13 motoneurons projecting to either part of the sartorius muscle maintained their phase-dependent selectivity without exception. For the phase of EMG recruitment associated with the unit discharge, the covariance of unit activity and EMG amplitude was just as high as for units projecting to unifunctional muscles, typically accounting for 88-95% of the variance of unitary spike rates.
Swing Task Group SA Motoneuron
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A c t i v i t y of a v e n t r a l r o o t axon p r o j e c t i n g t o SA-a muscle d u r i n g normal walking, t r a c e s a s i n Figure 1. Note u n i t a c t i v i t y only during swing phase EMG a c t i v i t y of parent muscle SA-a.
During r e f l e x responses e l i c i t e d by saphenous nerve s t i m u l a t i o n , a l l of t h e b i f u n c t i o n a l muscles (SA-a, SA-my and RF, which i s normally a c t i v e only during s t a n c e f o r slow walking) and t h e i r motor axons were r e c r u i t e d q u i t e homogeneously. S i x u n i t s s t u d i e d thoroughly under t h i s paradigm ( t h r e e from SA-a and t h r e e from RF) a l l responded with t y p i c a l f l e x o r muscle responses including s h o r t l a t e n c y (5-15 msec) and/or long l a t e n c y (20-35 msec) e x c i t a t o r y responses t h a t were complexly g a i t e d i n both t h e stance and swing phases of locomotion. I n p a r t i c u l a r , u n i t s p r o j e c t i n g t o b i f u n c t i o n a l muscles t h a t were normally a c t i v e only during t h e s t a n c e (extensor) phase produced r e f l e x r e s p o n s e s t h a t were t y p i c a l of f l e x o r muscles, whereas u n i t s p r o j e c t i n g t o pure extensor muscles ( v a s t i ) produced t y p i c a l extensor r e f l e x e s , t y p i f i e d by i n h i b i t i o n and occasional l a t e e x c i t a t o r y rebound. CONCLUSIONS These s t u d i e s have confirmed many important concepts of motor c o n t r o l derived from reduced p r e p a r a t i o n s , w h i l e a t t h e same time p o i n t i n g o u t important d e t a i l s and exceptions t h a t have not been seen i n such preparations : 1.
Motor u n i t s a r e r e c r u i t e d i n a r e g u l a r and reproducible manner, with recruitment order a t l e a s t l o o s e l y c o r r e l a t e d with t h e conduction v e l o c i t y of t h e motor axons ( s e e Henneman and Mendell, 1981, f o r
The main d i f f e r e n c e between e a r l y r e c r u i t e d (presumably slow t w i t c h , f a t i q u e r e s i s t a n t ) and l a t e r e c r u i t e d (high threshold, presumably f a s t , f a t i g u a b l e ) motor u n i t s i s i n t h e proportion of t h e time t h a t they a r e a c t i v e , r a t h e r t h a n i n t h e p a r t i c u l a r spike r a t e s o r
patterns generated. This is in contrast to earlier findings regarding 'tonic' vs. 'phasic' patterns reported for low and highthreshold units in reduced preparations (Granit et al., 1957), and it has important implications for theories regarding plasticity of muscle unit properties (see e.g. Salmons and Vrbova, 1969; reviewed by Burke, 1981b). The very close covariance between unit spike rates and EMG amplitude suggests that all members of each functional recruitment group receive the same net input signal, differing only in amplitude and/or threshold of response. Conversely, EMG signals recorded and processed in the manner employed here appear to provide a remarkably good indication of this command signal, including fine details of temporal modulation resulting from the interaction of the locomotor pattern generator and various descending and segmental influences. Bifunctional muscles such as anterior sartorius may be divided functionally into separately recruited groups, perhaps related to kinematically opposite conditions such as active lengthening during stance phase and active shortening during swing (see Loeb, 1985). The above-noted task-dependent recruitment groups can be over-ridden during different motor tasks such as the flexor reflex, where all units with a similar mechanical action (in this case, hip flexion) were recruited together regardless of their locomotor task group participation. REFERENCES
Abraham, L. D., Marks, W. B,, and Loeb, G. E., 1985, The distal hindlimb musculature of the cat: Cutaneous reflexes during locomotion,
pp.345-422, ~ashin~tbn, DC. Burke, R. E., 1981b, The stability of motor unit types in response to altered functional demand: Hypertrophy, atrophy and reinnervation
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Applications. Proc. of Workshop on Solid State Physical Sensors for Biomedical Application," M. R. Neuman et al., eds., CRC Press, Inc., Boca Raton. Salmons, S., and Vrbova, G., 1969, The influence of activity on some contractile characteristics of mammalian fast and slow muscles, J.Physiol.(Lond.), 201:535-549. Walmsley, B., Hodgson, J. A., and Burke, R. E., 1978, Forces produced by medial gastrocnemius and soleus muscles during locomotion in freely moving cats, J.Neurophysiol., 41:1203-1216. Zajac, F. E., and Young, J. L., 1980, Discharge properties of hindlimb motoneurons in decerebrate cats during locomotion induced by mesencephalic stimulation, J.Neurophysiol., 43:1221-1235.