European Journal of Neuroscience

European Journal of Neuroscience, Vol. 28, pp. 1427–1435, 2008 doi:10.1111/j.1460-9568.2008.06458.x

COGNITIVE NEUROSCIENCE

Bilateral motor resonance evoked by observation of

a one-hand movement: role of the primary motor cortex

Paola Borroni,1 Marcella Montagna,2 Gabriella Cerri2 and Fausto Baldissera2

1
Dipartimento di Medicina, Chirurgia e Odontoiatria, Università degli Studi di Milano, Via A.di Rudinı̀ 8, 20142 Milano, Italy
2
Istituto di Fisiologia Umana II, Università degli Studi di Milano, Milano, Italy

Keywords: human, mirror neurones, monosynaptic reflex, premotor cortex, transcranial magnetic stimulation

Abstract
In humans, observation of movement performed by others evokes a subliminal motor resonant response, probably mediated by the
mirror neurone system, which reproduces the motor commands needed to execute the observed movement with good spatial and
temporal fidelity. Motor properties of the resonant response were here investigated with the ultimate goal of understanding the
principles operating in the transformation from observation to internal reproduction of movement. Motor resonance was measured
as the modulation of excitability of spinal motoneurones, evoked by the observation of a cyclic flexion-extension of one hand. The
first two experiments showed that the observation of a one-hand movement always evoked a bimanual resonant response
independent of which hand was observed and that these bilateral responses were consistently phase-linked. H-reflexes
simultaneously recorded in right and left flexor carpi radialis muscles were always modulated ‘in-phase’ with each other. The goal of
the third experiment was to define the role of primary motor cortex in the bilateral resonant response. Bilateral H-reflexes were
recorded during a temporary inactivation induced by transcranial magnetic stimulation over the left cortical hand motor area of
observers. The finding that such cortical depression abolished the H-reflex modulation of only the right flexor carpi radialis
motoneurones, leaving it unchanged on the left side, suggested that both primary motor areas were activated by the premotor
cortex and transmit the resonant activation through crossed corticospinal pathways. The data provide further evidence that the
subliminal activation of motor pathways induced by movement observation is organized according to general rules shared with the
control of voluntary movement.

Introduction
Motor resonance in humans, the subliminal activation of motor
pathways induced by the mere observation of somebody else’s
movement, is thought to result from the activity of neurones
homologous to the mirror neurones described in the monkey ventral
premotor cortex (diPellegrino et al., 1992; Gallese et al., 1996).
A large number of studies, utilizing various imaging and recording
techniques, have demonstrated the activation of the human ventral
premotor cortex during hand action observation (Rizzolatti et al.,
2001; Rizzolatti & Craighiero, 2004), as well as the subliminal
modulation of both cortical and spinal hand motor pathways of human
observers (for a review see Fadiga et al., 2005).
The resonant response induced by observation of hand movements
generates a modulation of both H-reflex and motor evoked potentials
(MEPs) in the muscles producing the observed action (Fadiga et al.,
1995; Strafella & Paus, 2000; Borroni et al., 2005). This modulation
replicates the temporal profile and muscle selection of the motor
command that the observer would utilize if executing the same
Correspondence: Dr P. Borroni, as above.
E-mail: paola.borroni@unimi.it
Received 26 March 2008, revised 11 August 2008, accepted 15 August 2008
movement (Montagna et al., 2005). During the observation of a
flexion-extension hand movement, the H-reflex in flexor carpi radialis
(FCR) shows a facilitation maximum during flexion and minimum
during extension. Moreover, MEPs in antagonist muscles (flexor and
extensor carpi radialis) are modulated in phase opposition, reflecting
their natural reciprocal activation during execution of oscillations with
the hand (Borroni et al., 2005). Further experiments (Borroni &
Baldissera, 2008) have revealed that, in addition to the strict matching
of activation pattern during observation and execution, observing the
movement of one hand can also modulate motor pathways to the
contralateral hand.
Many questions still remain about the functional connections and
the specific rules that link and regulate the activation of the mirror
system to the activation of motor pathways. The evidence available so
far suggests that the resonant response originates in the ventral
premotor cortex (Nishitani & Hari, 2000; Avenanti et al., 2007) either
bilaterally or just in the left hemisphere and descends to spinal
motoneurones via the primary motor cortex (M1).
In the present study, two novel aspects of motor resonance are
explored. First, it is demonstrated that the resonant response develops
in muscles moving both hands during the observation of a one-hand
movement and the temporal relationship of the resonant responses

ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
1428 P. Borroni et al.
between hands and with the observed movement is described. Second,
attention is directed to the neural pathways involved in the bimanual
motor resonance. As the temporary inhibition of the left M1 eliminates
the FCR H-reflex resonant modulation on the right side (Borroni &
Baldissera, 2008), we investigated whether the same left M1
inactivation leaves the reflex modulation on the left side unaffected;
this result would indicate that the right and left M1s are activated
in parallel by the mirror system in one or both hemispheres.
Conversely, the loss of the motor resonant response on both sides
would suggest that it originates in the left hemisphere and is only
subsequently transmitted to both sides, either through commissural
connections between motor cortices or through bilateral projections of
the left M1.
Materials and methods
Experiments were performed with informed and consenting subjects,
according to the rules of the Declaration of Helsinki, and approved by
the local ethics committee (Comitato Etico dell’Università degli Studi
di Milano). Subject handedness was established using the standard
Edinburgh Handedness Inventory (Oldfield, 1971).
A total of 14 right-handed subjects (nine females and five males,
aged 18–48 years) participated in the three experiments: 11 subjects in
Experiment 1, two of these subjects plus three new ones in Experiment
2 and five subjects in Experiment 3. In all experiments, subjects
observed a series of four to five flexion-extension cycles of the prone
hand, performed in front of them by one of the experimenters (mover),
who followed the 1 Hz tempo of a metronome, heard through
headphones. The mover oscillating hand (either right or left depending
on the experiment) rested on a metal platform that could oscillate
around the wrist axis. The platform angular position was continuously
recorded with a Spectrol 534 1 kW potentiometer coaxial with its pivot
and digitized at 250 Hz. When, during the third oscillation, the
platform reached a preselected position in the cycle, a trigger signal
was released to activate the stimulator and elicit H-reflexes in the FCR
muscle of observers. H-reflexes were evoked by electrical stimulation
of the median nerve at the elbow (square pulse, 0.8 ms duration) and
recorded with bipolar surface electrodes placed on the FCR muscle
belly. The same electrodes were also used to confirm the lack of any
electromyographic (EMG) activity in this muscle during movement
observation. Signals were amplified, filtered (10–1000 Hz) and A ⁄ D
converted (5 kHz). All recorded signals (H-reflex and movement
traces) were stored and peak-to-peak amplitudes of H-reflexes were
measured. Amplitude of the FCR H-reflex at rest was maintained
between 5% and 15% of the maximal direct motor response.
Synchronous stimulation of the median nerve on the two sides was
used to elicit H-reflexes simultaneously in the right and left FCR
muscles of observers.
Preliminary experiments were carried out to verify that the
amplitude of FCR H-reflexes is not different when obtained during
simultaneous stimulation of both median nerves vs. stimulation of
each median nerve separately. Five subjects (different from the 14
subjects who participated in the main experiments) were prepared for
stimulation and recording on both sides and H-reflexes (20 right alone,
20 left alone, and 40 right and left together) were recorded in random
order. Data were analysed with a two-way, repeated measures anova
with H-reflex side (right or left) and condition (alone or together) as
within-subject factors. No significant main effect or interaction was
found. Mezzarane & Khon (2002) have also reported a lack of
interference when simultaneously evoking H-reflexes in the soleus
muscles of the two sides.
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
In the first two experiments a stimulus was delivered in each trial at
one of five fixed delays (0, 200, 400, 600 and 800 ms), dividing the
cycle period in equal parts; a total of 75 trials, 15 per delay, were
obtained in each subject. Delays were selected in semi-random order,
i.e. completing a cluster of all five delays before starting the new
random selection again. In order to eliminate long-term variability
independent of the stimulus position in the cycle, in each observer the
deviation (in lV) from the mean of the five responses recorded in each
cluster was calculated for each delay. This last value was then
averaged with those obtained at the same delay in the other clusters.
For each subject, the average of all observed hand movements was
calculated and fitted by a four-parameter (period, offset, amplitude and
phase) sinewave function. As cycle period varied among trials and
among movers by about 5% of its average value, normalization was
necessary in order to bring to unity movement records from different
trials and different movers. To maintain the temporal correlation
between the time courses of observed movement and response
modulation in observers, the same normalization was performed on
the five delays at which the H-reflexes were recorded (normalized
delays are indicated as d1–d5). Average reflex values obtained at the
same delay (see above) were assigned to the normalized delays.
Averages were fitted with a two-parameter sinewave function
(amplitude and phase, as after normalization period = 1 and offset = 0
for all). Finally, H-reflex modulation from all different subjects was
plotted, after normalization to each subject’s sinewave amplitude,
together with a grand-average movement trace in a single-cycle
diagram and fitted with a common two-parameter sinewave function.
In the third experiment the modulation of the FCR H-reflex was
tested during a transcranial magnetic stimulation (TMS)-induced
inhibition of the hand area of the left M1. The subject’s head was
restrained by a fitted support and a figure-of-eight-shaped coil,
connected to a magnetic stimulator (Magstim 200, maximal power 2.2
T), was placed, tangentially to the scalp, over the cortical focus for
TMS activation of hand muscles. The optimal position for eliciting
MEPs in the FCR muscle was located and marked on the subject’s
scalp. MEPs were monitored with the same surface electrodes used
during all experiments to record H-reflexes and electromyograms (see
above). Motor threshold at rest was defined as the minimum TMS
stimulus intensity that produced an MEP in the contralateral FCR
muscle at least three of six consecutive times. After the beginning of
the experiment, the effectiveness of TMS was regularly ascertained.
After verifying that TMS given during voluntary contraction
produced an EMG silent period lasting 60–70 ms, in the actual
experiment, in which there was no contraction of the FCR muscle, we
used the highest TMS intensity that did not produce MEPs at rest. In
this way a cortical inhibition without any spinal component was
obtained (Davey et al., 1994; Classen & Benecke, 1995; Reid et al.,
2002). The H-reflex excitability was tested during movement obser-
vation at 50 ms after subliminal TMS as well as without any TMS
conditioning.
For practical reasons, in the cortical inhibition experiments
H-reflexes were recorded at only two points during the observation
of the movement cycle, corresponding to the minimum (D1) and
maximum (D2) of the five-delay reflex modulation cycle, as previ-
ously determined for every observer (the full, five-delay data from the
five observers who participated in this experiments are included in
Fig. 1). A total of 30 TMS conditioned and unconditioned H-reflexes
were gathered for each subject, at each delay.
The significance of all best-fit sinewave regressions was ascertained
with a standard anova. A two-way anova was used to analyse data
from Experiments 1 to 3. A paired t-test was utilized to compare
H-reflex modulations in the single subject presented in Fig. 3 and
European Journal of Neuroscience, 28, 1427–1435

Fig. 1. Modulation of H-reflexes recorded simultaneously in right (A) and left
(B) FCR muscles during observation of one cycle of a flexion-extension
movement of the right hand (C, average movement ± SEM). (A and B)
Cumulative plots for 11 right-handed subjects, aligned after time and amplitude
normalization. Average data points from all subjects were fitted with a common
sinewave function (A and B, continuous lines), with the same period as that of
the average movement (C). D/, phase difference between reflex modulation
(vertical dashed line) in right and left flexor muscles of all observers and right
hand oscillation of the mover (vertical dotted line). Ext, upward direction of the
moving hand. Flex, downward direction of the moving hand. At the bottom is a
graphic representation of the experiment.
H-reflex amplitudes of all subjects in Fig. 4, with and without TMS
conditioning.
Results
In the present experiments the H-reflex excitability in the FCR
muscles of the resting right and left forearms was recorded simulta-
neously in subjects observing a cyclic flexion-extension movement of
one hand executed by a different individual. With this simple
movement, the same sinewave function can be used to fit both the
observed wrist oscillation and the resonance-induced reflex modula-
tion, thus quantifying the temporal correspondence between the events
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
European Journal of Neuroscience, 28, 1427–1435
Bilateral response to observation of one-hand movement 1429
and determining the precise phase relation of the left and right
modulations with the observed movement and with each other.
Bilateral resonant response during the observation
of movements of one hand
In the first experiment, H-reflex modulations in right and left FCR
muscles were recorded simultaneously in 11 right-handed subjects
observing the cyclic flexion-extension movement (Fig. 1C, sinewave
fitting, R2 = 0.99; P < 0.0001) executed with the right hand. Figure 1
also shows H-reflex modulations of all subjects pooled together and
fitted with a sinewave function with the same period as that obtained
for the movement [R2 = 0.48, P < 0.0001 for the right FCR (Fig. 1A);
R2 = 0.18, P < 0.002 for the left FCR (Fig. 1B)]. Note that the time
course of the H-reflex modulation in the observer’s right and left wrist
flexors closely reproduces that of the mover’s right hand movement,
with maximum H-reflex amplitude during flexion and minimum
during extension. Normalized H-reflex modulation values were
analysed by means of a two-way, repeated measures anova, with
delay (d1–d5) and H-reflex side (right or left) as within-subject factors.
A significant main effect was found for the delay factor (F4,40 = 4.02;
P = 0.0078), indicating that H-reflex modulation is significant for
FCR muscles on both sides. Intersubject variability of the modulation
amplitude appears greater for the left side reflex but no significant
interaction between factors was found, indicating that the reflex
modulation was the same in the left and right FCR muscles, when
subjects were observing the movement of the right hand.
The phase of the function fitting the H-reflex modulation best was
virtually identical on the two sides, i.e. 52 on the right and 59 on the
left. These values of the phase differences (D/) between the reflex
modulation and observed movement reflect those found between the
overt muscular contraction and the actual movement, when executed
by the observer with the hand in the same position and at the same
oscillation frequency (Borroni et al., 2005).
To evaluate whether observing the movement of the dominant hand
might have played an important role in determining the bimanual
resonant response, a second experiment was performed by simulta-
neous recording of the H-reflex modulation in the right and left FCR
muscles in five right-handed subjects observing the same movement
executed with the left hand. Results were very similar to those
obtained in the first experiment. Figure 2 shows that the time course of
the H-reflex modulation in both the right (Fig. 2A) and left (Fig. 2B)
wrist flexors of right-handed observers followed that of the observed
movement of the left hand (Fig. 2C). The sinusoidal fit of the reflex
modulations was again highly significant (Fig. 2A, R2 = 0.53,
P < 0.0001; Fig. 2B, R2 = 0.38, P < 0.0001). Normalized H-reflex
modulation values were analysed by means of a two-way, repeated
measures anova, with delay (d1–d5) and H-reflex side (right or left)
as within-subject factors. As for data in Fig. 1, a significant main effect
was found for the delay factor (F4,16 = 3.63, P = 0.0273), indicating
that H-reflex modulation is significant for FCR muscles on both sides.
Again, the intersubject variability of the modulation amplitude appears
greater for the left side reflex, this time during the observation of the
movement of the left hand but no interaction between factors was
found and thus no difference between the H-reflex modulations on the
left and right sides.
Note that phase differences (D/) between modulation and
movement are very consistent in both experiments, being 52 and
64 on the right, and 59 and 54 on the left in Experiments 1 and 2,
respectively. These results confirm that the laterality of the observed
hand (right or left) does not have an important influence in shaping
1430 P. Borroni et al.
Fig. 2. H-reflex modulation recorded simultaneously in right (A) and left (B)
FCR muscles during observation of one cycle of a flexion-extension movement
of the left hand (C, average movement ± SEM). (A and B) Cumulative plots for
five right-handed subjects, aligned after time and amplitude normalization.
Average data points from all subjects were fitted with a common sinewave
function (A and B, continuous lines) with the same period as that of the average
movement (C). D/, phase differences between reflex modulation (vertical
dashed lines) in right and left flexor muscles of all observers and left hand
oscillation of the mover (vertical dotted lines). Ext, upward direction of the
moving hand. Flex, downward direction of the moving hand. At the bottom of
the figure is a graphic representation of the experiment.
the bimanual resonant response and the strict phase correlation of the
reflex modulation in both hands reveals a strong bimanual coordi-
nation between hands, evoked by the observation of a one-hand
movement.
Neural pathways involved in the bilateral resonant response
The involvement of the M1s in the transmission of resonant activation
to bilateral spinal circuits was studied by evoking H-reflexes in FCR
muscles on both sides simultaneously, during the observation of a
flexion-extension movement of the right hand, while a TMS-elicited
cortical inhibition inactivated the left primary motor hand area of
observers. Reflexes were evoked only at the minimum (D1) and
maximum (D2) of the reflex modulation cycle, as previously
determined with a full, five-delay curve in each subject.
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
Results of this experiment are illustrated in Figs 3 and 4. Figure 3
shows H-reflexes recorded in one subject; the modulation induced by
movement observation in the control condition (reflexes not condi-
tioned with TMS) is evident in both right and left FCR muscles
(Fig. 3A and B), with reflexes being significantly larger in D2 than in
D1 (paired t-test: P = 0.0143, t = )2.3287 for the right side;
P = 0.0151, t = )2.5841 for the left side). In the control condition,
the modulation of the H-reflex in both right and left FCR muscles was
significantly different in D1 and D2 in each of the five subjects. When
the H-reflex was evoked at 50 ms from the TMS, i.e. during the
inhibitory conditioning of the left M1, the difference in reflex
amplitude disappeared on the right side (Fig. 3C, paired t-test,
P = 0.4107, t = )0.8316), whereas it remained significant on the left
(Fig. 3D, paired t-test, P = 0.0486, t = )2.3417).
Figure 4 shows the results of the five subjects pooled together;
the significant H-reflex modulation normally present on both sides
in the control condition (filled circles, Fig. 4A, right and Fig. 4B,
left) was abolished on the right side (Fig. 4C, empty circles) when
the reflex was evoked during the TMS-conditioned inhibition of the
left M1, whereas it remained unchanged on the left side (Fig. 4D,
empty circles). Left and right H-reflex modulations were analysed
separately as reflexes on the left and right sides are independent
measures obtained by separate stimulation of the two median
nerves, evoking responses in autonomous spinal circuits. This
assumption is also supported by the results of our preliminary
experiments, carried out to ensure that the amplitude of FCR
H-reflexes is not different when obtained during simultaneous
stimulation of both median nerves vs stimulation of each median
nerve separately (see Materials and methods). Thus, normalized
H-reflex modulation values were analysed by means of a two-way,
repeated measures anova, with delay (D1 and D2) and TMS
conditioning as within-subject factors. A significant main effect was
found for the delay factor, in both right and left FCR reflexes,
indicating an overall significant H-reflex modulation on both sides
(F1,4 = 35.00, P = 0.0041 for the right FCR; F1,4 = 26.64,
P = 0.0077 for the left FCR). In addition, a significant interaction
between delay and TMS conditioning (F1,4 = 88.29, P = 0.0007)
was found on the right but not on the left side (F1,4 = 5.67,
P = 0.0758). Posthoc analysis (Tukey HSD test) revealed that, in
the right FCR, muscle reflexes at D1 and D2 were significantly
different in the control (P = 0.000883) but not in the TMS
(P = 0.999100) condition.
The overall mean amplitudes of TMS conditioned and uncondi-
tioned reflexes were not different (average amplitudes ± SD: right
FCR control, 237 ± 94 lV and silent period, 229 ± 89 lV; left FCR
control, 285 ± 139 lV and silent period, 301 ± 109 lV; paired
t-test, P = 0.2604, t = 1.1276 on the right and P = 0.1165,
t = 1.5742 on the left), showing that TMS alone did not produce
any effect on the amplitude of the H-reflex itself. This methodo-
logical aspect is important because it verifies that TMS conditioning
per se does not change the amplitude of the H-reflex, i.e. that a
TMS pulse subliminal for producing MEPs and delivered with a
50 ms delay does not have any spinal inhibitory effect on the reflex
transmission, and confirms the conclusions of Experiment 3
regarding the cortical origin of the inhibition responsible for
eliminating the modulation of the right FCR H-reflex.
The fact that, in all subjects, during the inhibition of the left M1 the
resonant modulation of the H-reflex is depressed on the right
(contralateral) side but remains unaffected on the left (ipsilateral) side
supports the view that observation of a unilateral movement induces a
parallel activation of M1s on both sides and that each autonomously
drives the controlateral spinal neurones.
European Journal of Neuroscience, 28, 1427–1435
 Bilateral response to observation of one-hand movement 1431

Fig. 3. Average traces ± SD of 30 H-reflexes recorded simultaneously in the right and left FCR muscles of a single observer during observation of a flexion-
extension movement of the right hand (not shown) at delays corresponding with the subject’s minimum (D1) and maximum (D2) of the reflex modulation cycle
previously measured in that subject. Top panels (A, right hand and B, left hand) show the modulation of H-reflex not conditioned by TMS (control). (C) The
modulation of the right FCR H-reflex is eliminated during a cortical inhibition induced by TMS conditioning of the left M1 of the observer. (D) Such TMS
conditioning does not affect the FCR H-reflex modulation on the left. Time on the abscissa shows the latency of the H-reflex response (about 15 ms) from median
nerve stimulation (0 ms). At the bottom the graphic representation of the experiment indicates that, during TMS stimulation of the left primary cortex, motor
commands to the contralateral hand are temporarily silenced.

Discussion
The main result of the present experiments is that the observation of a
one-hand movement evokes the activation of a coordinated subliminal
motor command to both hands, which is transmitted bilaterally to
spinal motoneurones through their contralateral M1.
Bilateral resonant response during the observation
of movements of one hand
In summary, the excitability of motoneurones innervating wrist flexor
muscles (FCR) of both sides was sinusoidally modulated by
observation of a sinusoidal flexion-extension movement of one hand,
irrespective of whether the right or left hand was observed. More
importantly, the modulation was synchronous on the two sides and the
rising phase to maximal facilitation coincided with the flexion phase of
the observed movement.
Such a bilateral involvement indicates that motor resonance is not
limited to a one-to-one correspondence with the observed moving
hand but spreads to the other hand, even though it is not included in
the observed movement. This is in line with earlier experiments in the
monkey, in which single mirror neurones in the ventral premotor
cortex (in both hemispheres) were shown to respond to the observation
of movement by either hand of the experimenter (Gallese et al., 1996);
these neurones were also active during movement of either hand of the
monkey, consistent with the existence of abundant bilateral premotor-
to-motor cortical projections (Matsumura & Kubota, 1979; Muakkassa
& Strick, 1979; Ghosh et al., 1987) and with the activation of the
premotor cortex (along with other cortical areas) during bimanual
movements (Baraldi et al., 1999; Binkofski et al., 1999; Rizzolatti &
Luppino, 2001; Swinnen, 2003).
Although it is possible that the premotor cortex is engaged bilaterally
in motor resonance during observation of either left or right hands
because it does not code the laterality of the observed hand, but a more
abstract representation of the movement, the present data do not allow
testing of this hypothesis. Rather, having recorded the excitability of a
specific spinal motor neuron population, driving a single muscle (the
FCR) directly involved in the execution of the observed wrist flexion, it
may be more useful to interpret our results in the context of the known
organization and control of explicit motor acts. In humans, bilateral
activation of homologous upper limb muscles seems to regularly occur
during voluntary movements of one single hand or arm. Thus, the
coordinated bilateral modulation revealed by the present experiments
could also be a consequence of the neural links normally involved in the

ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
European Journal of Neuroscience, 28, 1427–1435
1432 P. Borroni et al.

Fig. 4. Panels are organized as in Fig. 3. The top panels show the H-reflex modulation recorded in five subjects at two delays corresponding to the minimum (D1)
and maximum (D2) of their reflex modulation cycle. H-reflexes were recorded simultaneously in the right (A) and left (B) FCR muscles during observation of a
flexion-extension movement of the right hand (symbolized by the thin dotted lines). Data points of different subjects were aligned after time and amplitude
normalization. The modulation of the right FCR H-reflex is eliminated during a cortical inhibition induced by TMS conditioning of the left M1 of observers (C),
whereas it remains unchanged in the left FCR H-reflex (D). In all panels triangles are averages of the five subjects (±SD).

execution of coordinated bimanual tasks. Indeed, our results agree with
the common principle that ‘in-phase’ coupling is the preferred and most
stable mode of coordination during the execution of many bimanual
tasks, particularly rhythmic oscillations (for a review see Swinnen,
2003). Furthermore, the synergy of muscular activation may change
from subliminal to supraliminal according to the postural context. In
comfortably sitting subjects, during the execution of rhythmic oscilla-
tions of the wrist, activation in the contralateral resting FCR is
subliminal, as revealed by the amplitude modulation of both the H-reflex
and the TMS elicited MEPs observed by Carson et al. (2004).
Ridderikhoff et al. (2005) also found evidence for bursting activity in
the resting extensor carpi radialis, phase-linked with the extension of the
contralateral wrist, and Baldissera et al. (2008) observed that, in standing
subjects who used the left hand for postural fixation, flexion-extension
movements of the right hand in the horizontal plane are accompanied by
frank EMG activation of the contralateral homologous muscles.
In this light, the present finding that motor resonance involves
muscles on both sides suggests that the ‘mirror imitation’ is a
subliminal replica not only of the explicit actions on which
observation is focused but of the whole activation pattern utilized
during execution, including not directly observable muscular syner-
gies and anticipatory postural adjustments occurring in other limbs.
This conclusion is also supported by preliminary experiments in which
a resonant response is evoked in hand flexor muscles upon observation
of a flexion-extension movement of the foot (Borroni et al., 2006).
Neural pathways involved in the bilateral resonant response
The second series of experiments showed that temporary inhibition of
the left M1 eliminates the excitability modulation of spinal motoneu-
rones innervating the right FCR muscle, leaving the resonant response
unchanged on the left side.
Had the modulation been eliminated on both sides, two conclusions
would have been possible: (i) the resonant command travels from the
left premotor cortex to the left M1, then to the other side through
commissural connections between M1s and then to spinal circuits or,
less likely, (ii) it travels to the left M1 only, which in turn modulates
spinal motoneurones on both sides via both contra- and ipsilateral
corticospinal projections. As, however, modulation of the FCR
motoneurones remained unaltered on one (left) side, we conclude
that bilateral activation does not depend on commissural connections
between the M1s and that, after activation from the premotor cortex,
each M1 modulates the excitability of its spinal motoneurones via
crossed corticospinal projections. As the TMS-induced cortical
inhibition should affect neurones giving origin to both crossed and
ipsilateral corticospinal pathways, the fact that the H-reflex modula-
tion disappears on the right side and is maintained on the left indicates
that the resonant effects are transmitted by the crossed, not by the
ipsilateral, corticospinal projection. In turn, this suggests that, during
the resonance phenomena, the premotor cortex ⁄ cortices activate(s) in
parallel the M1s of the two sides.
At variance with the above conclusions about the role of the M1,
subliminal excitation of spinal motoneurones during action observa-
tion might, at least theoretically, also be achieved through direct
premotor projections to the spinal cord (Dum & Strick, 2002).
However, the involvement of the M1 in mediating the motor resonant
response is supported by several lines of evidence. First suggested by
the strong anatomical and physiological connections between premo-
tor and motor cortices, particularly abundant for the hand area,
described in both monkey and humans (Geyer et al., 2000; Munchau
et al., 2002; Cerri et al., 2003; Shimazu et al., 2004; Umiltà et al.,

ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
European Journal of Neuroscience, 28, 1427–1435

2007; Boros et al., 2008; Schmidlin et al., 2008), it has recently
gathered much support from motor resonance studies on human
subjects. Experiments showing that MEPs induced by TMS in the
motor cortex are modulated by hand movement observation (Fadiga
et al., 1995; Gangitano et al., 2001; Aziz-Zadeh et al., 2002) have
suggested a role for the primary motor area in motor resonance; this
role has been confirmed by further TMS experiments in which care
was taken to separate the cortical and spinal components of the MEP
response modulation (Strafella & Paus, 2000; Montagna et al., 2005).
Although activation of M1 is not demonstrated by neuroimaging
techniques (not surprisingly, in the absence of overt movement)
(Rizzolatti et al., 1996; Iacoboni et al., 1999; Perani et al., 2001),
magneto- and electroencephalographic recordings have shown that M1
is activated in both hemispheres during the observation of unilateral
(Hari et al., 1998; Nishitani & Hari, 2000) and bilateral (Cochin et al.,
1999) hand movements. Finally, the involvement of M1 in the
transmission of motor resonance to spinal circuits was more directly
revealed by experiments in which the left M1 hand area was
temporarily inactivated by TMS conditioning, resulting in the loss of
the resonant H-reflex modulation in the right FCR muscle (Borroni &
Baldissera, 2008).
There are a few considerations about the use of TMS-induced cortical
inhibition in the present experiments. This temporary depression of
cortical activity can be measured as the silencing of voluntary EMG
activity in contralateral muscles after a TMS stimulus (silent period) and
is due to the after-hyperpolarization refractoriness and postsynaptic
inhibition that follow the firing of both cortical neurones and spinal
motoneurones (Mills, 1999; Reid et al., 2002). It can also be measured in
paired-pulsed TMS experiments, which show that the response to the
second pulse is inhibited from about 30 to 200 ms following the first
pulse, irrespective of whether the target muscle is contracted or at rest
(Wassermann et al., 1996). If, as done here, the TMS intensity is
maintained subthreshold for inducing an MEP in contralateral muscles,
and thus no spinal motoneuronal firing is produced, the remaining
inhibition can be considered to be exclusively the result of cortical
processes (Davey et al., 1994; Classen & Benecke, 1995).
In addition to the development of a contralateral inhibition, a much
shorter inhibitory effect has also been described in ipsilateral muscles,
thought to be mediated by intracortical circuits that inhibit transcallosal
excitatory fibres connecting the two M1s (Avanzino et al., 2007;
Trompetto et al., 2004). This ipsilateral silent period is normally evoked
with TMS stimulations of intensity well above threshold for inducing a
contralateral MEP (Wassermann et al., 1991; Ferbert et al., 1992;
Avanzino et al., 2007; Trompetto et al., 2004). Thus, an ipsilateral
inhibition is reasonably not evoked in our stimulation paradigm, as
subthreshold TMS intensities were always used; even in the unlikely
event that some minimal ipsilateral inhibitory effect might have been
produced, the H-reflex modulation was measured at a conditioning
interval of 50 ms from the TMS, which is likely to be beyond the
duration of any ipsilateral effect. In any event, the result itself, i.e. the fact
that the reflex modulation remained unchanged on the side ipsilateral to
TMS stimulation, indicates that no interhemispheric cortical inhibition
was interfering with the development of the resonant response.
The results of brain imaging studies in human subjects have been
somewhat inconsistent with respect to the lateralization of activation of
the premotor cortex during observation of unilateral hand movements.
Some studies show an almost equal bilateral activation of the ventral
premotor cortices (Buccino et al., 2001, 2004; Iacoboni et al., 2005),
whereas others report a stronger or even exclusive activation in the left
hemisphere (Iacoboni et al., 1999; Buccino et al., 2004; Gallagher &
Frith, 2004; Tai et al., 2004). The present study shows that both
hemispheres are involved in shaping the motor resonant response, at
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
European Journal of Neuroscience, 28, 1427–1435
Bilateral response to observation of one-hand movement 1433
least at the level of the M1 but, given the abundant bilateral projections
from premotor to motor cortices (Matsumura & Kubota, 1979;
Muakkassa & Strick, 1979; Ghosh et al., 1987), it does not resolve the
question relative to the hemisphere of origin. Addressing this specific
point, a recent study (Fecteau et al., 2005) reports that motor resonance
in a callosotomized patient can be induced by the observation of both
the ispilateral and contralateral hand but remains localized in the left
hemisphere. This observation supports the view that motor resonance
might originate in the left premotor cortex and from there spread to
both M1s.
What complicates matters is that results of different studies are not
always fully comparable due to the great variability in complexity of the
observed actions and to the diversity of the context in which they are
performed. Just to mention one example from a strictly motor point of
view, the left hemisphere appears to have a special role in visually
guided precision grasping but not power grasping tasks (Gonzales et al.,
2006; Vainio et al., 2006). At a more abstract level, given the fact that the
left ventral premotor cortex in humans coincides with, or at least
includes, Broca’s area (Petrides et al., 2005), the stronger activation of
the left hemisphere reported by some authors could be interpreted in the
context of the hypothesized role of the mirror neuron system in language
acquisition (Rizzolatti & Arbib, 1998; Rizzolatti, 2005). Indeed, motor
resonance has often been reported to develop preferentially in the left
hemisphere when the observed actions had a communicative value,
whereas other non-communicative actions evoked a response in both
hemispheres (Buccino et al., 2004; Gallagher & Frith, 2004). Alterna-
tively, a predominant response of the left premotor cortex might have
been explained by the fact that, in most studies, observing subjects are
invariably right-handed and ⁄ or are usually observing movements
performed with the right hand. Our results suggest that, during the
observation of a flexion-extension movement of either hand, the
resonant response does not develop strictly or even better in the right
hand of right-handed observers or in the hand corresponding to the
observed one but rather it always develops as a coordinated bimanual
response, similar to the bilateral activation normally occurring during
voluntary movements of a single hand.
With the present experiments we hope to begin bridging the gap
between the more abstract functions of the mirror neuron system and its
more ‘basic’ motor functions, with the ultimate goal of defining and
understanding the neural properties of the system, which necessarily
underlie and constrain its function(s). From our results we conclude that:
(i) observation of a specific hand movement evokes the subliminal
implementation of the full motor programme necessary for that
movement; (ii) the motor programme includes both hands and (iii) it
is the simplest, most common one, i.e. the one that couples hands ‘in-
phase’; and (iv) M1 is activated on both sides, reaching spinal
motoneurones by way of crossed corticospinal projections. We interpret
the existence of features common to both motor resonance and the
control of voluntary movement in the light of our working hypothesis,
i.e. when a movement is executed explicitly or ‘mirrored’ subliminally,
the same rules apply. With this approach we intend to provide plausible
evidence that the observation of a given movement, which activates the
mirror system, is transformed into a specific subliminal motor activity
according to the same principles that organize the generation of
voluntary commands for that same movement.
Acknowledgements
This study was supported by grants from the Ministero dell’Università e della
Ricerca and by the Università degli Studi di Milano (PRIN 2006). We thank
Professor Alberto Morabito for his valuable advice on the statistical analysis of
the present data.
1434 P. Borroni et al.
Abbreviations
EMG, electromyographic; FCR, flexor carpi radialis; M1, primary motor
cortex; MEP, motor evoked potential; TMS, transcranial magnetic stimulation.
References
Avanzino, L., Teo, T.H. & Rothwell, J.C. (2007) Intracortical circuits modulate
transcallosal inhibition in humans. J. Physiol., 583, 99–114.
Avenanti, A., Bolognini, N., Maravita, A. & Aglioti, S.A. (2007) Somatic and
motor components of action simulation. Curr. Biol., 17, 2129–2135.
Aziz-Zadeh, L., Meada, F., Zaidel, E., Mazziotta, J. & Iacoboni, M. (2002)
Lateralization in motor facilitation during action observation: a TMS study.
Exp. Brain Res., 144, 127–131.
Baldissera, F., Rota, V. & Esposti, R. (2008) Anticipatory postural adjustments
in arm muscles associated with movements of the contralateral limb and their
possible role in interlimb coordination. Exp. Brain Res., 185, 63–74.
Baraldi, P., Porro, C.A., Serafini, M., Pagnoni, G., Murari, C., Corazza, R. &
Nichelli, P. (1999) Bilateral representation of sequential finger movements in
human cortical areas. Neurosci. Lett., 269, 95–98.
Binkofski, F., Buccino, G., Posse, S., Seitz, R.J., Rizzolatti, G. & Freund, H.-J.
(1999) A fronto-parietal circuit for object manipulation in man: evidence
from an fMRI-study. Eur. J. Neurosci., 11, 3276–3286.
Boros, K., Poreisz, C., Munchau, A., Paulus, W. & Nitsche, M.A. (2008)
Premotor transcranial direct current stimulation (tDCS) affects primary motor
excitability in humans. Eur. J. Neurosci., 27, 1292–1300.
Borroni, P. & Baldissera, F. (2008) Activation of motor pathways during
observation and execution of hand movements. Social Neurosci., 3, 276–
288.
Borroni, P., Montagna, M., Cerri, G. & Baldissera, F. (2005) Cyclic time course
of motor excitability modulation during the observation of a cyclic hand
movement. Brain Res., 1065, 115–124.
Borroni, P., Cerri, G., Montagna, M. & Baldissera, F. (2006) The observation of
a single limb movement induces multi-limb motor resonance in the observer.
Acta Physiol., 188(Suppl. 652), 77.
Buccino, G., Binkofski, F., Fink, G.R., Fadiga, L., Fogassi, L., Gallese, V.,
Seitz, R.J., Zilles, K., Rizzolatti, G. & Freund, H.-J. (2001) Action
observation activates premotor and parietal areas in a somatotopic manner:
an fMRI study. Eur. J. Neurosci., 13, 400–404.
Buccino, G., Vogt, S., Ritzl, A., Fink, G.R., Zilles, K., Freund, H.-J. &
Rizzolatti, G. (2004) Neural circuits underlying imitation learning of hand
actions: an event related fMRI study. Neuron, 42, 323–334.
Carson, R.G., Riek, S., Mackey, D.C., Meichenbaum, D.P., Willms, K., Forner,
M. & Byblow, W.D. (2004) Excitability changes in human corticospinal
forearm projections and spinal reflex pathways during rhythmic voluntary
movement of the opposite limb. J. Physiol., 560, 929–940.
Cerri, G., Shimazu, H., Maier, M.A. & Lemon, R.N. (2003) Facilitation from
ventral premotor cortex of primary motor cortex outputs to macaque hand
muscles. J. Neurophysiol., 90, 832–842.
Classen, J. & Benecke, R. (1995) Inhibitory phenomena in individual motor
units induced by transcranial magnetic stimulation. Electroencephal. Clin.
Neurophysiol., 97, 264–274.
Cochin, S., Barthelemy, C., Roux, S. & Martineau, J. (1999) Observation and
execution of movement: similarities demonstrated by quantified electroen-
cephalography. Eur. J. Neurosci., 11, 1839–1842.
Davey, N.J., Romaiguere, P., Maskill, D.W. & Ellaway, P.H. (1994) Suppres-
sion of voluntary motor activity revealed using transcranial magnetic
stimulation of the motor cortex in man. J. Physiol., 477, 223–235.
Dum, R.P. & Strick, P.L. (2002) Motor areas in the frontal lobe of the primate.
Physiol. Behav., 77, 677–682.
Fadiga, L., Fogassi, L., Pavesi, G. & Rizzolatti, G. (1995) Motor facilitation
during action observation: a magnetic stimulation study. J. Neurophysiol.,
73, 2608–2611.
Fadiga, L., Craighiero, L. & Olivier, E. (2005) Human motor cortex excitability
during the perception of others’ action. Curr. Opin. Neurobiol., 15, 213–218.
Fecteau, S., Lassonde, M. & Théoret, H. (2005) Modulation of motor cortex
excitability during action observation in disconnected hemispheres. Neuro-
report, 16, 1591–1594.
Ferbert, A., Priori, A., Rothwell, J.C., Day, B.L., Colebatch, J.G. & Mardsen,
C.D. (1992) Interhemispheric inhibition of the human motor cortex.
J. Physiol., 453, 525–546.
Gallagher, H.L. & Frith, C.D. (2004) Dissociable neural pathways for the
perception and recognition of expressive and instrumental gestures. Neuro-
psychologia, 42, 1725–1736.
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
Gallese, V., Fadiga, L., Fogassi, L. & Rizzolatti, G. (1996) Action recognition
in the premotor cortex. Brain, 119, 593–609.
Gangitano, M., Mottaghy, F.M. & Pascual-Leone, A. (2001) Phase-specific
modulation of cortical motor output during movement observation. Neuro-
report, 12, 1489–1492.
Geyer, S., Matelli, M., Luppino, G. & Zilles, K. (2000) Functional neuro-
anatomy of the primate isocortical motor system. Anat. Embryol., 202, 443–
474.
Ghosh, S., Brinkman, C. & Porter, R. (1987) A quantitative study of the
distribution of neurones projecting to the precentral motor cortex in the
monkey (M. fascicularis). J. Comp. Neurol., 259, 424–444.
Gonzales, C.L.R., Ganel, T. & Goodale, M.A. (2006) Hemispheric specializa-
tion for the visual control of action is independent of handedness.
J. Neurophysiol., 95, 3496–3501.
Hari, R., Forss, N., Avikainen, S., Kirveskari, S., Salenius, S. & Rizzolatti, G.
(1998) Activation of human primary motor cortex during action observation:
a neuromagnetic study. Proc. Natl Acad. Sci. USA, 95, 15061–15065.
Iacoboni, M., Woods, R.P., Brass, M.P., Bekkering, H., Mazziotta, J.C. &
Rizzolatti, G. (1999) Cortical mechanisms of human imitation. Science, 286,
2526–2528.
Iacoboni, M., Molnar-Szakacs, I., Gallese, V., Buccino, G., Mazziotta, J.C. &
Rizzolatti, G. (2005) Grasping the intentions of others with one’s own mirror
neuron system. PLoS Biol., 3, 529–535.
Matsumura, M. & Kubota, K. (1979) Cortical projections to hand-arm motor
area from post-arcuate area in macaque monkeys: a histological study of
retrograde transport of horseradish peroxidase. Neurosci. Lett., 11, 241–246.
Mezzarane, R.A. & Khon, A.F. (2002) Bilateral soleus H-reflexes in humans
elicited by simultaneous trains of stimuli: symmetry, variability, and
covariance. J. Neurophysiol., 87, 2074–2083.
Mills, K.R. (1999) Magnetic Stimulation of the Human Nervous System.
Oxford University Press, Oxford, NY.
Montagna, M., Cerri, G., Borroni, P. & Baldissera, F. (2005) Excitability
changes in human corticospinal projections to muscles moving hand and
fingers while viewing a reaching and grasping action. Eur. J. Neurosci., 22,
1513–1520.
Muakkassa, K. & Strick, P. (1979) Frontal lobe inputs to primate motor cortex:
evidence for four somatotopically organized premotor areas. Brain Res., 177,
176–182.
Munchau, A., Bloem, B.R., Irlbacher, K., Trimble, M.R. & Rothwell, J.C.
(2002) Functional connectivity of human premotor and motor cortex
explored with repetitive transcranial magnetic stimulation. J. Neurosci., 22,
554–561.
Nishitani, N. & Hari, R. (2000) Temporal dynamics of cortical representation
for action. Proc. Natl Acad. Sci. USA, 97, 913–918.
Oldfield, R.C. (1971) The assessment and analysis of handedness: the
Edinburgh Inventory. Neuropsychologia, 9, 97–113.
diPellegrino, G., Fadiga, L., Fogassi, L., Gallese, V. & &. Rizzolatti, G. (1992)
Understanding motor events: a neurophysiological study. Exp. Brain Res.,
91, 176–180.
Perani, D., Fazio, F., Borghese, N.A., Tettamanti, M., Ferrari, S., Decety, J. &
Gilardi, M.C. (2001) Different brain correlates for watching real and virtual
hand actions. NeuroImage, 14, 749–758.
Petrides, M., Cadoret, G. & Mackey, S. (2005) Orofacial somatomotor
responses in the macaque monkey homologue of Broca’s area. Nature, 435,
1235–1238.
Reid, A.E., Chiappa, K.H. & Cros, D. (2002) Motor threshold, facilitation and
the silent period in cortical magnetic stimulation. In Pascual-Leone, A.,
Davey, N.J., Rothwell, J., Wasswermann, E.M. & Puri, B.K. (Eds), Handbook
of Transcranial Magnetic Stimulation. Arnold, London, pp. 97–111.
Ridderikhoff, A., Peper, C.L. & Beek, P.J. (2005) Unraveling interlimb
interactions underlying bimanual coordination. J. Neurophysiol., 94, 3112–
3125.
Rizzolatti, G. (2005) The mirror neuron system and its function in humans.
Anat. Embryol., 210, 419–421.
Rizzolatti, G. & Arbib, A. (1998) Language within our grasp. TINS, 21, 188–194.
Rizzolatti, G. & Craighiero, L. (2004) The mirror-neuron system. Annu. Rev.
Neurosci., 27, 169–192.
Rizzolatti, G. & Luppino, G. (2001) The cortical motor system. Neuron, 31,
889–901.
Rizzolatti, G., Fadiga, L., Matelli, M., Bettinardi, V., Paulesu, E., Perani, D. &
Fazio, F. (1996) Localization of grasp representation in humans by PET: 1.
observation versus execution. Exp. Brain Res., 111, 246–252.
Rizzolatti, G., Fogassi, L. & Gallese, V. (2001) Neurophysiological mecha-
nisms underlying the understanding and imitation of action. Nat. Rev.
Neurosci., 2, 661–669.
European Journal of Neuroscience, 28, 1427–1435

Schmidlin, E., Brochier, T., Maier, M.A., Kirkwood, P.A. & Lemon, R.N.
(2008) Pronounced reduction of digit motor responses evoked from macaque
ventral premotor cortex after reversible inactivation of the primary motor
cortex hand area. J. Neurosci., 28, 5772–5783.
Shimazu, H., Maier, M.A., Cerri, G., Kirkwood, P.A. & Lemon, R.N.
(2004) Macaque ventral premotor cortex exerts powerful facilitation of
motor cortex outputs to upper limb motoneurons. J. Neurosci., 24, 1200–
1211.
Strafella, A.P. & Paus, T. (2000) Modulation of cortical excitability during
observation: a transcranial magnetic stimulation study. Neuroreport, 11,
2289–2292.
Swinnen, S.P. (2003) Intermanual coordination from behavioural principles to
neural-network interactions. Nat. Rev. Neurosci., 3, 350–361.
Tai, Y.F., Scherfler, C., Brooks, D.J., Sawamoto, N. & Castiello, U. (2004) The
human premotor cortex is ‘mirror’ only for biological actions. Curr. Biol.,
14, 117–120.
ª The Authors (2008). Journal Compilation ª Federation of European Neuroscience Societies and Blackwell Publishing Ltd
European Journal of Neuroscience, 28, 1427–1435
Bilateral response to observation of one-hand movement 1435
Trompetto, C., Bove, M., Marinelli, L., Avanzino, L., Buccolieri, A. &
Abruzzese, G. (2004) Suppression of the transcallosal motor output: a
transcranial magnetic stimulation study in healthy subjects. Exp. Brain Res.,
158, 133–140.
Umiltà, M.A., Brochier, T., Spinks, R.L. & Lemon, R.N. (2007) Simultaneous
recording of macaque premotor and primary motor cortex neuronal
populations reveals different functional contributions to visuomotor grasp.
J. Neurophysiol., 98, 488–501.
Vainio, L., Ellis, R., Tucker, M. & Symes, E. (2006) Manual asymmetries in
visually primed grasping. Exp. Brain Res., 173, 395–406.
Wassermann, E.M., Fuhr, P., Cohen, L.G. & Hallett, M. (1991) Effects of
transcranial magnetic stimulation on ipsilateral muscles. Neurology, 41,
1795–1799.
Wassermann, E.M., Samli, A., Mercuri, B., Ikoma, K., Oddo, D., Grill, S.E. &
Hallett, M. (1996) Responses to paired transcranial magnetic stimuli in
resting, active and recently activated muscles. Exp. Brain Res., 109, 158–163.