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Journal of Neuroscience Methods 90 (1999) 47 – 55
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A comparison of cross-correlation and surface EMG techniques


used to quantify motor unit synchronization in humans
John G. Semmler, Michael A. Nordstrom *
Department of Physiology, Uni6ersity of Adelaide, Adelaide, SA 5005, Australia

Received 10 March 1999; accepted 8 May 1999

Abstract

Two methods used to estimate the strength of motor unit (MU) synchronization in a muscle are the direct cross-correlation of
MU discharge times, and averaging of the surface electromyogram (SEMG) with respect to discharge of a reference MU.
Although indirect, the latter approach has the advantage that a global estimate of MU synchrony can be obtained quickly and
easily. The two methods are generally regarded as providing equivalent information on the extent of MU synchronization in a
muscle, but this proposition has not previously been tested quantitatively. In the present study, we used both the SEMG technique
(189 MUs) and cross-correlation of MU discharge (498 MU pairs) to estimate MU synchrony in 28 first dorsal interosseus (FDI)
muscles from 16 subjects. Despite considerable overlap in the identity of MUs used to quantify synchrony with each method,
linear regression revealed no significant correlation between the estimates of MU synchronization in FDI muscles obtained with
the two techniques (r 2 =0.04, n=28). This discrepancy was not due to insufficient sampling of the MU population with the
cross-correlation method, although we found evidence for a non-uniform tendency for synchronous discharge in two of 13 motor
units providing sufficient data for the analysis. The most likely explanation for the discrepancy between the estimates of MU
synchrony is that methodological problems with the SEMG technique limit its accuracy. These problems are difficult to avoid
under normal experimental conditions, and we conclude that the SEMG method is not reliable for quantitative comparisons of
MU synchrony between muscles and subjects. © 1999 Elsevier Science B.V. All rights reserved.

Keywords: Cross-correlation; Motor unit; Surface electromyogram; Corticospinal tract; First dorsal interosseous; Synchrony

1. Introduction histogram constructed of the peri-event discharge times


of the other MU (Moore et al., 1966). Although this
During a voluntary isometric contraction there is an direct method accurately quantifies synchronization of
increased tendency for motor units (MUs) to discharge MU pairs, in practice it poses considerable technical
within a few milliseconds of each other more often than difficulties. Short-term synchrony is fairly weak, and
is expected due to chance. This MU synchronization is trials of several minutes duration are needed to provide
of interest because its features can be used to infer sufficient counts for a reliable estimate of synchrony
details of the operation of last-order neurons with from the cross-correlogram. It is difficult to identify the
widely divergent projections to motoneurons (Sears and same MUs in the intramuscular recordings from several
Stagg, 1976; Datta and Stephens, 1990). The corti- electrodes for long periods, as their action potential
cospinal projection appears to be of major importance waveforms may change. A further complication is that
for MU synchronization in distal hand muscles (Datta two intramuscular electrodes are needed, and each must
et al., 1991; Farmer et al., 1993). MU synchrony can be sample from a separate population of MUs to avoid
quantified directly by cross-correlation of individual spurious effects in the cross-correlogram due to non-
discharge times of MU pairs, in which the discharge random discrimination errors arising from action po-
times of one MU are used as a reference, and a tential superimpositions involving an MU detected in
both channels. The large territory occupied by muscle
* Corresponding author. Tel.: +61-8-8303-4567; fax: + 61-8-8303- fibres of a single MU make this difficult to avoid in
3356. small muscles. Finally, it is necessary to sample from a

0165-0270/99/$ - see front matter © 1999 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 5 - 0 2 7 0 ( 9 9 ) 0 0 0 6 9 - 2
48 J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55

number of MU pairs to obtain a reliable estimate of the years). Features of MU discharge, MU synchrony and
overall extent of MU synchrony in a muscle, because force tremor in these subjects have been reported previ-
the strength of MU synchrony varies over a wide range ously (Semmler and Nordstrom, 1995, 1998). The ex-
for different MU pairs in the same muscle (Bremner et tensive MU sample obtained in FDI muscles in these
al., 1991; Nordstrom et al., 1992). subjects made the present investigation possible. As the
The technical challenges of the cross-correlation tech- extent of FDI MU synchrony differs in the three
nique have prompted the use of alternative methods groups (Semmler and Nordstrom, 1998) these subjects
which are quicker and easier to use. Milner-Brown et exhibit a large range of MU synchrony in FDI, which
al. (1973) introduced a technique for estimating the improves the likelihood of demonstrating a significant
global MU synchronization in a muscle which consists correlation between estimates of MU synchrony ob-
of comparing the simultaneously recorded unrectified tained with the two methods.
and full-wave rectified surface EMG (SEMG) which The experimental arrangement and protocol for
have been averaged with respect to the discharge of a recording the surface EMG and MU discharge proper-
reference MU. Comparison of the areas of the rectified ties have been described previously (Semmler and
and unrectified SEMG averages (above baseline levels) Nordstrom, 1995, 1998). Briefly, the subjects attended
provides an index of synchronization of reference MU the laboratory on two or more separate occasions,
discharges with those of other active MUs. The SEMG where SEMG and MU activity were recorded from one
synchronization index derived using this method has hand on each occasion. The subjects’ right or left arm
the advantage that all active MUs make some contribu- and hand were secured in a manipulandum, and the
tion to the SEMG signal, and so an analysis of a force of abduction was measured by a load cell which
relatively short period of discharge of the reference MU was aligned with the distal interphalangeal joint of the
provides a global estimate of its synchronization with index finger. The SEMG of FDI was recorded with
other MUs in the muscle. Synchronization indices can bipolar Ag/AgCl electrodes placed 2–3 cm apart. MU
be quickly and easily obtained for a number of refer- activity was recorded simultaneously with two separate
ence MUs in a single session. Using this method, for fine-wire electrodes (1–2 cm interelectrode distance)
example, Milner-Brown et al. (1975) have reported that which were inserted percutaneously into the FDI with a
strength-training enhances MU synchrony in the first 25-gauge disposable needle. Myoelectric signals were
dorsal interosseous (FDI) muscle, a change which has amplified (1000× ), filtered (bandwidth 2 Hz–10 kHz)
been attributed to strengthening of transcortical reflex and recorded on FM tape (Vetter model 400D, Rebers-
pathways. burg, PA, USA, 22 kHz/ch) for off-line analysis. The
It is generally accepted that the SEMG and MU DC force signal was filtered (0–50 Hz) and digitised (1
cross-correlogram techniques provide equivalent infor- kHz) on-line on a Macintosh computer.
mation on the extent of MU synchronization in a To begin the experiment the subjects performed a
muscle. Surprisingly, however, this proposition has not steady, low-force, isometric abduction of the index
been quantitatively tested using experimental data. The finger. A single MU was chosen by the experimenters
SEMG technique has recently been shown to be subject for the subject to control at a comfortable discharge
to technical problems which raise questions about its rate (feedback unit). The subjects were provided with
reliability for estimating MU synchronization (Yue et audio and visual feedback of MU discharge on an
al., 1995). In the present study we have obtained spike- oscilloscope screen. The subject’s task was to control
train data from an extensive sample of MUs in 28 FDI the mean firing rate of the feedback unit at a constant
muscles, and used these data to estimate MU syn- target level (usually 8, 10 or 12 Hz) for 1–5 min. The
chrony in these muscles with both the SEMG technique activity of additional MUs was monitored during the
(189 MUs) and cross-correlation of MU discharge (498 trial to confirm that discriminable MU potentials were
MU pairs). The results show that the two methods do present in each channel for off-line cross-correlation.
not produce equivalent estimates of the overall extent The procedure was repeated following repositioning of
of MU synchronization in a muscle. both electrodes, in order to sample from as many
different MUs as possible in the session.
All analyses were performed off-line from the taped
2. Methods records. Single MUs from each intramuscular electrode
were discriminated using a computer-based template-
All subjects volunteered to participate in the study matching algorithm (SPS 8701, Signal Processing Sys-
and gave informed consent to the procedures, which tem, Malvern, S.A., Australia). Action potentials
were approved by the Human Research Ethics Com- belonging to a particular MU were identified on the
mittee at the University of Adelaide. MU activity was basis of waveform shape, and great care was taken to
recorded from 28 FDI muscles in five musicians, five confirm the identity of units discriminated during dif-
strength-trained and six untrained subjects (ages 18–47 ferent trials (see Nordstrom et al., 1992). ISIs of iden-
J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55 49

tified MUs were measured (9250 ms resolution) using was above the baseline activity (black and hatched area,
an in-built function of the SPS 8701 and stored on Fig. 1B and C), and above the voltage levels which
computer. ISI records were scrutinised for every trial occurred in the unrectified EMG trace (hatched area
and each discriminated MU to assess discrimination only, Fig. 1B and C). The strength of MU synchroniza-
accuracy. ISI histograms were constructed from the tion was determined by computing the ratio of the
discharge times of each MU. ISI files with \ 5% dis- synchronous area (black area, Fig. 1B and C) to the
crimination error (usually missed spikes due to super- area of the unrectified EMG within the peak region
impositions) were excluded from all analyses. (dotted and hatched area, Fig. 1B and C).
For 189 MUs, TTL pulses corresponding to the time MU synchronization was also assessed using cross-
of discharge of a single MU discriminated with the SPS correlation of MU discharge times (Nordstrom et al.,
8701 were sent to a second computer with the corre-
1992). MUs detected with separate electrodes in the
sponding surface EMG (2 kHz sampling rate). The
single MU discharge times were used as the reference
(t =0 ms) for spike-triggered averaging (STA) of the
SEMG. STA of the digitally full-wave rectified and
unrectified EMG was performed with a custom-de-
signed computer program. Each STA had a duration of
9 85 ms around the time of discharge of the reference
MU, and was based on 150 reference MU discharges.
A comparison of the area of the unrectified and
rectified EMG averages was used to provide an esti-
mate of the strength of MU synchronization within the
MU population. This method has previously been de-
scribed by Milner-Brown and co-workers (Milner-
Brown et al., 1973, 1975) and is shown for two MUs in
Fig. 1B and C. Briefly, STA of the unrectified and
full-wave rectified SEMG was performed for each refer-
ence MU. The unrectified SEMG average essentially
represents the contribution of only the reference MU to
the SEMG, because the contribution of the positive and
negative waveform components from other MUs not
synchronized to the reference MU average to zero. The
rectified SEMG average provides a measure of the total
electrical activity in the muscle around the time of
reference MU discharge, which includes the waveform Fig. 1. Estimation of the strength of MU synchronization from the
of the reference unit after rectification, an average cross-correlogram and the corresponding SEMG average. (A) Esti-
EMG level (background) due to independent firing of mation of MU synchrony using the cross-correlogram of MU dis-
charge. The lower trace is the cross-correlogram of the discharge
other MUs, an artifact associated with signal rectifica-
times of two concurrently active MUs in FDI. The mean bin count of
tion (due to a partial summation of the rectified wave- off-peak bins was 9.7 (horizontal line). This value served to distin-
form and the background activity; Milner-Brown et al. guish between the counts expected due to chance (vertically hatched
(1973); see also Yue et al., 1995) and the contribution area) from those counts in excess of chance (black area) in the peak
of other MUs that are synchronized to the reference region. The position and duration of the synchronous peak was
judged visually from the cumulative sum (CUSUM, upper trace). In
MU. For each reference MU, the unrectified (with
this example, peak duration (vertical dashed lines) was 19 ms centred
negative phases inverted) and rectified EMG averages at t = −2 ms. The synchronization index CIS for this MU pair was
were superimposed and the boundaries for the area of 1.21 extra counts s − 1. (B, C) Estimation of MU synchrony using the
synchronous activity were identified (dashed vertical SEMG spike-triggered averaging method (n = 150) for the two MUs
lines, Fig. 1B and C). This consisted of detecting the in (A). The area of the unrectified EMG average is represented by the
first location either side of the central peak, starting at dotted and hatched area. The area of the rectified EMG average is
represented by the black and hatched area. The duration of the peak
the point of MU discharge (arrow, Fig. 1B and C), is designated by the dashed vertical lines. The extent of MU synchro-
where the rectified EMG level was equal to the mean nization is calculated from the ratio of the synchronized area (black
EMG level due to the asynchronous firing of other area) to the total area of the unrectified EMG average within the
MUs (i.e. background EMG level; dashed horizontal peak region (dotted and hatched area). The arrow indicates the time
lines, Fig. 1B and C). From within the peak region, the of discharge of the reference MU. (B) The SEMG synchronization
index for this MU is 0.31 with a peak duration of 19 ms. (C) Data
area due to the synchronous firing of other MUs was from the second concurrently active MU in (A). This MU was used
established (black area in Fig. 1B and C). This area as the reference MU in the cross-correlogram (A). SEMG synchro-
represented the area in the rectified EMG trace which nization index for this MU was 1.03 with a peak duration of 24 ms.
50 J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55

same trial were paired for cross-correlation (498 MU


pairs). All cross-correlation histograms had 1-ms bin
widths and spanned a period 100 ms before and after
the discharge of the reference unit (Fig. 1A). His-
tograms with a mean bin-count B4 were not analysed
further. The position and duration of the synchronous
peak was judged visually through the use of the cumu-
lative sum procedure (CUSUM; Ellaway (1978); dotted
vertical lines, Fig. 1A). The significance of synchronous
peaks in the cross-correlogram was assessed using the
method described by Wiegner and Wierzbicka (1987).
A standard peak duration of 11 ms, centred at time
zero, was used for quantification of the strength of
synchrony in the MU pair if no significant peak from
Fig. 2. Relationship between the estimates of the strength of motor
the cross-correlogram was identified. The strength of
unit synchronization in FDI obtained using the surface electromyo-
MU short-term synchronization was quantified using gram (SEMG synchrony index) and the cross-correlogram (synchrony
the synchronization index CIS (common input index CIS). Data from 28 FDI muscles showing the mean SEMG
strength), which is the number of synchronous dis- synchrony index for each muscle (mean seven reference MUs per
charges in the cross-correlogram in excess of chance muscle, range 2 – 15) plotted against the mean MU synchrony index
CIS (mean 18 MU pairs per muscle, range 7 – 30) for the correspond-
(dark area, Fig. 1A) divided by the duration of the trial,
ing muscle. Linear regression revealed no significant correlation be-
i.e. the frequency of extra synchronous discharges in tween these two estimates of MU synchrony in FDI (r 2 =0.04).
the MU pair (Nordstrom et al., 1992).
Data are presented as mean9 S.E. unless otherwise Mean (9 S.D.) duration of MU synchrony in the 28
stated. Linear regression was used to assess the rela- FDI muscles was 15.79 3.2 ms when estimated using
tionships between estimates of MU synchronization the 291 cross-correlograms with significant central
obtained with the two methods. Statistical significance peaks (an average of 10 cross-correlograms per muscle,
was regarded as PB0.05.
range 1–23). The mean duration of MU synchrony in
these FDI muscles estimated with the SEMG method
3. Results
was 16.8 93.4 ms (189 MUs). The two estimates of the
mean duration of short-term synchrony in these mus-
The extent of MU synchrony in FDI was assessed
cles were not significantly different (paired t-test; P\
using the SEMG synchronization index for 189 MUs
0.05, n= 28). There was no significant correlation
from 28 FDI muscles in 16 subjects. MU synchroniza-
between the estimates of synchronous peak duration
tion was also assessed from cross-correlation of dis-
obtained by the two methods (r 2 = 0.095, n= 28, Fig.
charge of 498 MU pairs in the same muscles, which
3).
included the 189 MUs used for the SEMG analysis.
The relationship between the estimates of MU syn- The estimates of overall synchrony in FDI muscles
chrony in FDI obtained with the two techniques is reported in Figs. 2 and 3 were obtained using a rela-
shown for 28 muscles in Fig. 2. The mean strength of tively large sample of MUs for studies of this type (an
MU synchronization in FDI muscles estimated using average per muscle of seven reference MUs for the
the SEMG method varied over a 10-fold range (range SEMG index and 18 MU pairs for the cross-correlo-
0.12 –1.15), with an average of seven (range 2–15) grams). Nevertheless, in view of the poor correlation
reference MUs per muscle used to calculate the mean between the two estimates of synchrony in FDI, it is
SEMG synchronization index. The mean strength of necessary to consider whether the MU sample in each
MU synchronization (CIS) using cross-correlation of muscle was sufficiently large to provide a reliable esti-
MU discharge varied over a 200-fold range (range mate of the overall extent of MU synchrony in the
0.005–1.03 extra counts s − 1), with an average of 18 muscle. If some FDI MUs have consistently very weak
(range 7–30) MU pairs per muscle used to calculate the or strong synchronization with many other MUs, such
mean synchronization index CIS. With both techniques a non-uniformity in the distribution of synchronization
the estimates of the mean strength of MU synchroniza- within the MU population would increase the possibil-
tion varied over a large range in different muscles. For ity that under-sampling could produce errors in the
the 28 muscles, however, linear regression revealed no estimates of global synchronization in the muscles
significant correlation between the two measures of which may have obscured the expected positive correla-
MU synchronization (r 2 =0.04, Fig. 2). The two meth- tion between the two synchronization indices in Fig. 2.
ods clearly do not provide equivalent estimates of the Sampling error is more likely to be an issue for the
overall strength of MU synchrony in FDI. cross-correlogram analyses, because with the SEMG
J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55 51

technique all active MUs contribute to the SEMG


signals used to calculate the synchronization index for
each reference MU.
While it is common to encounter a MU pair with
unusually weak, or strong, synchronization in any mod-
erate-sized sample of the MU population of a muscle, it
is not known whether this represents a chance varia-
tion, or a consistent tendency for one or both of these
MUs to exhibit unusually weak or strong synchroniza-
tion with many or all MUs in the muscle. To address
this question, it is necessary to consider the strength of
synchrony exhibited by a single MU when paired with
many other concurrently active MUs, and compare this
with the overall extent of synchrony revealed in cross-
correlograms from a large sample of other MUs in that
muscle. Sufficient data for such a comparison are
difficult to obtain, but in our sample there were 13
MUs (from 10 FDI muscles) that were paired for
cross-correlation with at least five other concurrently
active MUs. For each of these 13 selected MUs, we Fig. 4. Comparison using cross-correlation of the strength of MU
calculated a measure of that MU’s synchronization synchrony for a restricted sample of MUs and a global estimate of
with other concurrently active MUs by pooling CIS MU synchrony in FDI. Abscissa: data from 13 selected FDI MUs
values from all available cross-correlograms for which which were cross-correlated with at least five other concurrently
active MUs. The mean synchrony index CIS from the restricted
that MU acted as the reference (this ranged between 5
sample of cross-correlograms involving the selected reference MU is
and 13, with an average of 7). This was compared with plotted against the mean synchrony index CIS from all remaining
the mean CIS obtained by pooling data from cross-cor- cross-correlograms for that FDI muscle (ordinate). The restricted
relograms involving all other MU pairs in that FDI sample was a reasonably good predictor of the global estimate of FDI
muscle (i.e. excluding pairs involving the selected MU). MU synchrony (linear regression line shown: r =0.86; P B0.001).
Filled symbols show the two selected MUs with significantly higher
This ranged between 9 and 23 cross-correlogram pairs, synchronization in their cross-correlograms than the global estimate
with an average of 14. The results of this analysis for for that FDI muscle.
the 13 MUs are summarised in Fig. 4.
Fig. 4 shows for each of 13 selected MUs, the mean obtained by pooling data from all other MU pairs in
synchronization index CIS for cross-correlograms with the muscle. Linear regression of these data revealed a
that unit as the reference, plotted against the mean CIS significant positive correlation (r= 0.86; PB0.001).
This indicates that data from a single reference MU
obtained using five or more cross-correlograms can be
used to provide a reasonable estimate of overall MU
synchrony in the FDI muscle. In two of 13 cases (Fig.
4, filled circles), however, the mean synchronization of
the selected reference unit with other MUs was signifi-
cantly different from the overall estimate of MU syn-
chrony obtained from cross-correlation of all other
available MU pairings (t-tests with Bonferroni correc-
tion for multiple contrasts). In one case, the mean9
S.E. (n) CIS of cross-correlograms involving the
selected MU was 0.7590.18 (7) versus 0.319 0.05 (23)
for all other MU pairings in that muscle (PB 0.05). In
the second case, mean CIS values were 0.46 9 0.05 (7)
Fig. 3. Relationship between the duration of the central synchronous for pairings with the selected MU versus 0.1590.02
peak in FDI obtained with the two methods. Data for 28 FDI (17) for all other MU pairings in the muscle (PB
muscles showing the mean duration of the central synchronous peak 0.001). These data suggest that a minority of MUs are
using the SEMG method plotted against the mean duration of the more highly synchronized than the rest of the motoneu-
significant synchronous peaks in the cross-correlograms (total 291
MU pairs) obtained from MUs in these muscles. Linear regression
ron pool, although in general MU synchrony is rela-
revealed no significant correlation between these two estimates of the tively uniformly distributed amongst the MU pool in
duration of MU synchrony in FDI (r 2 = 0.05). FDI.
52 J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55

The preceding analyses (Fig. 4) have established that pairs of MUs in FDI that exhibited large differences in
the overall extent of MU synchrony in FDI when the SEMG synchrony index (0.3 or greater), and for
quantified by pooling cross-correlation data from many which there were at least two cross-correlograms avail-
MU pairs is reasonably well estimated by sampling one able for each MU of the pair (sum total of cross-correl-
reference MU that is cross-correlated with at least five ograms involving the two units ranged from 5 to 12,
other MUs. For 11 of this sample of 13 MUs, the with a mean of 8). These selection criteria resulted in 20
pattern of MU synchrony was consistent with the be- MU pairs for the analysis. If the two methods of
haviour of a larger sample of MUs in the muscle. If the quantifying MU synchrony were related, it was ex-
SEMG index is a reliable estimate of the overall extent pected that the MU of the pair with the higher SEMG
of synchrony between the reference unit and other synchrony index would tend to have the higher mean
concurrently active MUs in the muscle, one would synchronization in the cross-correlograms. This pattern
expect a significant positive correlation between the was observed in only nine of 20 comparisons. In the
mean CIS estimated from cross-correlograms involving remaining 11 pairs, the MU with the higher SEMG
synchrony index had the lower mean CIS of the pair.
the 13 MUs selected for the Fig. 4 analysis, and the
This distribution of mean CIS differences for the 20
SEMG synchronization index calculated for the same
MU pairs was not significantly different from that
13 MUs. These data are shown in Fig. 5. There was no
expected due to chance (sign test; P = 0.83), so once
significant correlation between the two synchrony mea-
again for a group of MUs it was not possible to
sures obtained using these 13 MUs as the reference
demonstrate a relationship between the estimates of
(r = 0.24; P\0.05). When all available data were used
MU synchrony obtained with the SEMG and cross-cor-
for the regression analysis (184 MUs for which a relation methods.
SEMG synchrony index was calculated, and the MU
was paired with at least one other MU for an estimate
of synchrony from the cross-correlogram) the relation- 4. Discussion
ship between the SEMG synchrony index and mean
CIS was not significant (r =0.14, P =0.06). Implicit in the use of the SEMG synchronization
An additional comparison was used to assess the index is the assumption that it provides similar infor-
relationship between the two methods of quantifying mation on the extent of MU synchronization in a
MU synchrony for a population of MUs. We selected muscle as the cross-correlation technique, yet this
proposition had not previously been tested quantita-
tively. Fig. 2 shows that estimates of the strength of
MU synchrony in FDI muscles obtained using the two
techniques were not related in any meaningful way,
despite a wide range of values for both measures in the
28 FDI muscles studied and considerable overlap in the
identity of MUs used to quantify synchrony with each
measure. This suggests that either one or both of the
techniques are not providing an accurate measure of
MU synchrony in the muscle. Both techniques seem to
be reflecting short-term synchronization, as the mean
duration of the synchronous peaks was brief (approxi-
mately 16 ms), and estimates of peak duration obtained
with the two techniques were not significantly different
(Fig. 3).
The discrepancy between the estimates of MU syn-
chronization obtained with the two methods is nicely
illustrated by the data from two MUs shown in Fig. 1.
For this MU pair, cross-correlation revealed a high
degree of synchronization (Fig. 1A, CIS of 1.21). The
Fig. 5. Relationship between estimates of MU synchrony obtained SEMG synchronization index for one of these units was
with SEMG and cross-correlation techniques for 13 FDI MUs. Data rather low (Fig. 1B, 0.31) and for the other unit it was
from 13 FDI MUs selected because they were cross-correlated with at rather high (Fig. 1C, 1.03). One interpretation of these
least five other concurrently active MUs. For each MU, the SEMG data is that the discharge of this pair of MUs is highly
synchrony index is plotted against the mean synchronization index
CIS from all cross-correlograms involving that MU. There was no
synchronized, but the MU with the low SEMG syn-
significant correlation between the two estimates of MU synchrony chronization index (Fig. 1B) has weak synchrony with
obtained using these 13 reference MUs. the majority of other active MUs in FDI, while the unit
J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55 53

with the high SEMG index (Fig. 1C) is consistently ence MU did not differ significantly from the mean
strongly synchronized with other MUs. If MU syn- synchronization index obtained by pooling cross-corre-
chrony is non-uniformly distributed within FDI, this lation data from all other available MU pairings in that
would raise some concerns about the sample size muscle (Fig. 4). There was a significant positive correla-
needed to obtain a good estimate of the overall strength tion (r= 0.86) between the two synchrony measures
of MU synchrony in a muscle. An alternative explana- obtained from the restricted and larger sample of cross-
tion is that the high SEMG synchrony index for the correlograms (Fig. 4). However, two of 13 MUs studied
MU depicted in Fig. 1C is artifactual, and arises from were significantly more synchronized with other MUs
technical limitations of the SEMG technique (cf. Yue et than the global average obtained from all other MU
al., 1995). One problem seen here is that this MU’s pairings in the muscle. This indicates that there is not a
representation in the SEMG signal is small, and barely completely uniform distribution or effectiveness of
sufficient to exceed the baseline average rectified EMG shared, branched-axon inputs to all motoneurons in the
level (compare the size of the diagonally hatched areas pool; a minority (approximately 15% in our sample) of
in Fig. 1B and C). This poor signal-to-noise ratio will motoneurons share more common inputs than is the
lead to a high value for the synchronization index, even norm for their motoneuron pool. This finding supports
with a constant contribution to the peak in the rectified the view that the previously observed large variation in
EMG average from synchronized MUs (Yue et al., the strength of MU synchrony for individual MU pairs
1995). in a single FDI muscle (Bremner et al., 1991; Nord-
strom et al., 1992) does not arise by chance.
4.1. Sampling issues and the uniformity of MU The Fig. 4 analysis indicates that a reasonable esti-
synchronization for different MUs in a muscle mate of the overall extent of MU synchrony in FDI can
be obtained by cross-correlating the discharge of one
Both techniques rely on sampling a proportion of reference MU with as few as five other MUs. This
active MUs in FDI to estimate the overall extent of procedure provided an estimate of MU synchrony that
MU synchrony in the muscle. The human FDI has was positively correlated with the global estimate of
about 120 MUs (Feinstein et al., 1955). All MUs synchrony, and not significantly different from it in
included in the present study had recruitment force 85% of comparisons. As the average number of MU
thresholds B4 N, which is equivalent to about 10–15% pairs/muscle used to provide a global estimate of FDI
of FDI maximal voluntary force. It is difficult to esti- MU synchrony from the cross-correlograms was 18 for
mate how many MUs are active in FDI at these low the Fig. 2 analysis (minimum seven), and these did not
force levels, but it is likely to be a small fraction of the include the same reference MU in every pairing, we feel
total, as the abduction force during the trials was justified to conclude that the number and diversity of
commonly between 0.5 and 1 N. The mean number of MU pairs sampled per muscle was sufficient to provide
reference MUs per muscle used for the SEMG syn- a reliable global estimate of the strength of FDI MU
chrony analyses was seven, and the mean number of synchronization with the cross-correlation technique.
cross-correlograms analysed per muscle was 18. The The 13 MUs that could be cross-correlated with five
MU sample was considered to be reasonable for the or more MUs were used for a direct comparison of the
SEMG technique, in view of the fact that all active synchrony measures obtained with the SEMG and
MUs potentially contribute to the SEMG synchrony cross-correlation methods (Fig. 5). Even in this ‘best-
index obtained for each reference MU. The number of case’ comparison, the synchrony measures obtained
MU pairs used for the cross-correlogram analyses is a with the two techniques were not significantly corre-
more limited proportion of the possible combinations lated. Similarly, a comparison using 20 MU pairs with
of active MUs, and for this reason we looked for large differences in the SEMG synchrony index re-
evidence on the sample size needed to provide a reason- vealed that the MU of the pair with the higher SEMG
able estimate of overall MU synchronization using the synchrony index had no significant tendency to exhibit
cross-correlation technique. Lower limits on the sample higher mean synchrony in the cross-correlograms with
size required for a valid estimate of overall MU syn- other MUs.
chrony in a muscle, and the issue of the uniformity of Our data consistently show that measures of MU
the distribution of MU synchrony within the MU pop- synchrony obtained using the SEMG and cross-correlo-
ulation, have not been explored previously. This is gram techniques do not bear any useful relationship to
presumably because sufficient data for the comparisons one another. Our evidence suggests that these disparate
are difficult to obtain. findings are not due to under-sampling of the MU
In 11 of 13 (85%) cases in which an MU was paired population, despite a tendency for non-uniform syn-
with at least five other concurrently active MUs for chronization within a sub-set of the MU population in
cross-correlation analysis, the mean synchronization in- FDI. We now turn to consideration of methodological
dex for the restricted sample involving a common refer- limitations of the SEMG method.
54 J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55

4.2. Technical limitations of the surface EMG method that these were likely to be so difficult to achieve under
for quantifying MU synchrony normal experimental conditions that use of the method
to accurately assess MU synchrony was not practical.
The SEMG technique is an indirect estimate of MU The first was that the SEMG technique only be used
synchrony, with the complication that contributions of when the rectification artifact was negligible, which in
single MUs to the SEMG are not equal (depending on their simulation occurred when the peak rectified EMG
the size and number of muscle fibres, and their location signal in the average was more than three times as large
and orientation with respect to the recording elec- as the background EMG level. In the present study,
trodes). An important limitation of the SEMG method only 12 of 189 MUs (range 1–6 reference MUs per
is that, on theoretical grounds (Milner-Brown et al., muscle) in six muscles satisfied this rather stringent
1973), slightly positive synchronization ratios would be criterion. For these MUs, there was no significant
expected under asynchronous conditions. This is due to relationship between their SEMG synchrony index and
the partial summation of the rectified waveform and the the mean synchronization index CIS from all cross-cor-
background EMG activity, and is termed the rectifica- relograms (ranging 1–3) involving the MU (r=0.52,
tion artifact. This problem was recognised originally by P\ 0.05). The rectified EMG peak was more than
Milner-Brown et al. (1973) who used a theoretical twice as large as the background EMG level for only 43
approach to calculate the contribution of the rectifica- MUs in the present study. For these 43 MUs there was
tion artifact that accounted for changes in the signal-to- also no significant relationship between SEMG syn-
noise ratio. When the rectification artifact is calculated chrony index and the mean synchronization index CIS
and subtracted from the synchronization index, the from all cross-correlograms (ranging 1–10) involving
SEMG technique has been shown to provide a sensitive the MU (r= 0.17, P\ 0.05).
measure of the level of MU synchronization within a The second recommendation of Yue et al. (1995) was
population of neurons (Roscoe et al., 1985). However, that comparisons between different muscles should only
due to the extra data processing required to calculate be made for contractions involving a quite narrow
the rectification artifact, Milner-Brown et al. (1975) range of forces. This was because of the dependence of
chose to simplify the method by assuming that the the SEMG synchronization index on the level of muscle
rectification artifact was fixed, and was independent of excitation (presumably because recruitment influences
the signal-to-noise ratio. This simplified SEMG method the number of active MUs available to contribute to
was used by these authors to show an increase in MU the global synchrony measure). In their study of 56
synchronization with strength-training (Milner-Brown FDI MUs tested at two different levels of muscle
et al., 1975), a result that is widely cited in the literature activation, Yue et al. (1995) found that the surface
as a neural adaptation to strength training (Sale, 1987). synchrony index was 31% higher when the average
Technical problems associated with the SEMG muscle contraction force increased from about 4 to 7%
method have recently been highlighted by Yue et al. of MVC. Most of the index finger abduction forces at
(1995). Using experimental data from FDI MUs made which FDI MUs were studied in the present study were
to discharge under low- and high-rate conditions, Yue in the range 0.1–5% MVC, with no contraction over
et al. (1995) showed that the SEMG synchrony index 12% MVC. It is likely that differences in the number of
was significantly positively correlated with the level of active MUs over the force range at which different
EMG activity in the muscle. This was attributed to the MUs were tested were responsible for considerable
recruitment of additional synchronized MUs at higher variation in the SEMG synchronization indices in the
levels of muscle activation. Furthermore, using a com- present study.
puter simulation of 100 FDI MUs discharging asyn- In summary, there is a poor correlation between
chronously, Yue et al. (1995) reported that the global estimates of MU synchronization in FDI mus-
rectification artifact contributed from 0 to 0.15 to the cles using MU cross-correlation and SEMG methods.
SEMG synchronization index, and this figure varied Cross-correlation of MU discharge times is a more
non-linearly with the signal-to-noise ratio (defined in direct, and therefore more reliable measure of MU
this case as the ratio of the peak amplitude of the synchronization, provided that sufficient MU pairs are
rectified average relative to the average background included in the analysis. Although the tendency for
EMG level). The signal-to-noise ratio was reduced at synchronous discharge was higher than average for a
higher levels of background EMG activity, and as the minority (15%) of MUs, we have presented evidence
signal-to-noise ratio decreased the accuracy of the that our sample size was sufficient to obtain a reliable
SEMG index as an estimate of MU synchronization global estimate of MU synchrony in FDI using cross-
declined. correlation. The most likely explanation for the poor
Two recommendations were made by Yue et al. correlation between the estimates of MU synchrony
(1995) to minimise errors when using the SEMG obtained with the two methods is technical limitations
method to estimate MU synchrony, but they predicted of the SEMG method (Yue et al., 1995), which are
J.G. Semmler, M.A. Nordstrom / Journal of Neuroscience Methods 90 (1999) 47–55 55

difficult to overcome under normal experimental condi- Feinstein B, Lindegard B, Nyman E, Wohlfart G. Morphological
tions. We conclude that the simplified SEMG method studies of motor units in normal human muscles. Acta Anat
1955;23:127 – 42.
(Milner-Brown et al., 1973, 1975) is not reliable for Milner-Brown HS, Stein RB, Yemm R. The contractile properties of
quantitative comparisons of MU synchrony between human motor units during voluntary isometric contractions. J
muscles and subjects. Physiol (Lond) 1973;228:285 – 306.
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Acknowledgements Moore GP, Perkel DH, Segundo JP. Statistical analysis and func-
tional interpretation of neuronal spike data. Ann Rev Physiol
This work formed part of the PhD studies of JGS, 1966;28:493 – 522.
who was supported by a University of Adelaide Post- Nordstrom MA, Fuglevand AJ, Enoka RM. Estimating the strength
graduate Research Scholarship. MAN was an R.D. of common input to human motoneurons from the cross-correlo-
gram. J Physiol (Lond) 1992;453:547 – 74.
Wright Fellow of the NH&MRC of Australia.
Roscoe DD, Hamm TM, Reinking RM, Stuart DG. Detection of
synchrony in the discharge of a population of neurons. II. Imple-
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