Francesco Bolzoni

Evidence for long‐lasting subcortical facilitation by transcranial direct current stimulation in the cat

•  Transcranial constant current polarization of the human brain is on the increase in neurological practice because it improves several motor and cognitive functions of the human nervous system and because it is non‐invasive and technically simple. •  Here we show that transcranial brain polarization in anaesthetized animals not only affects cortical neurons, as is often assumed, but also facilitates activation of neurons in all investigated subcortical motor systems. •  In addition, the subcortical facilitation greatly outlasts (by at least hours) the period of transcranial polarization. These findings provide new evidence of plasticity at subcortical levels, the mechanisms for which remain to be investigated. •  In clinical practice, the subcortical effects of transcranial polarization may thus make an essential contribution to the beneficial effects of the treatment of motor impairments.

Subcortical effects of transcranial direct current stimulation in the rat

•  Previously demonstrated facilitation of activation of subcortical neurons by transcranial direct current stimulation (tDCS) in acute experiments on deeply anaesthetized animals was fairly weak. It resulted in only small increases in the amplitude and in a slight shortening of latencies of subcortically initiated descending volleys. •  Here we show that despite weak effects on descending volleys, EMG responses evoked in neck muscles by reticulospinal and rubrospinal neurons in deeply anaesthetized non‐paralysed rats are potently facilitated by tDCS and that the facilitation outlasts tDCS. •  We further show that the facilitatory subcortical effects of tDCS in the rat are evoked by cathodal rather than anodal polarization, i.e. by a polarity that is the reverse of that most often found to be effective in humans and in the cat. Anodal polarization depressed activation of the same rat subcortical neurons. •  These findings should assist further studies of mechanisms of tDCS in vivo in rodents.

Presynaptic and postsynaptic effects of local cathodal DC polarization within the spinal cord in anaesthetized animal preparations

Trans‐spinal DC stimulation affects both postsynaptic neurons and the presynaptic axons providing input to these neurons. In the present study, we show that intraspinally applied cathodal current replicates the effects of trans‐spinal direct current stimulation in deeply anaesthetized animals and affects spinal neurons both during the actual current application and during a post‐polarization period. Presynaptic effects of local cathodal polarization were expressed in an increase in the excitability of skin afferents (in the dorsal horn) and group Ia afferents (in motor nuclei), both during and at least 30 min after DC application. However, although the postsynaptic facilitation (i.e. more effective) activation of motoneurons by stimuli applied in a motor nucleus was very potent during local DC application, it was only negligible once DC was discontinued. The results suggest that the prolonged effects of cathodal polarization are primarily associated with changes in synaptic transmission.

Transcranial direct current stimulation of SMA modulates anticipatory postural adjustments without affecting the primary movement.

Recent works provide evidences that anticipatory postural adjustments (APAs) are programmed with the prime mover recruitment as a shared posturo-focal command. However the ability of the CNS to adjust APAs to changes in the postural context implies that the postural and voluntary components should take different pathways before reaching the representation of single muscles in the primary motor cortex. Here we test if such bifurcation takes place at the level of the supplementary motor area (SMA). TDCS was applied over the SMA in 14 subjects, who produced a brisk index-finger flexion. This activity is preceded by inhibitory APAs, carved in the tonic activity of Biceps Brachii and Anterior Deltoid, and by an excitatory APA in Triceps Brachii. Subjects performed a series of 30 flexions before, during and after 20 min of tDCS in CATHODAL, ANODAL or SHAM configuration. The inhibitory APA in Biceps and the excitatory APA in Triceps were both greater in ANODAL than in SHAM and CATHODAL configurations, while no difference was found among the latter two (ANODAL vs. SHAM: biceps +26.5%, triceps +66%; ANODAL vs. CATHODAL: biceps +20.5%, triceps: +63.4%; for both muscles, ANOVA p<0.02, Tukey p<0.05). Instead, the APA in anterior deltoid was unchanged in all configurations. No changes were observed in prime mover recruitment and index-finger kinematics. Results show that the SMA is involved in modulating APAs amplitude. Moreover, the differential effect of tDCS observed on postural and voluntary commands suggests that these two components of the motor program are already separated before entering SMA.

Accuracy of pointing movements relies upon a specific tuning between anticipatory postural adjustments and prime mover activation

Equilibrium‐perturbing forces associated with a voluntary upper‐limb movement can be strong enough to displace the whole‐body centre of mass. In this condition, anticipatory postural adjustments (APAs), developing in muscles other than the prime mover, are essential in maintaining the whole‐body balance. Here, we test the hypothesis that APAs preceding an upper‐limb target‐reaching movement could play a role also in controlling the movement accuracy.

Evidence that some long-lasting effects of direct current in the rat spinal cord are activity-independent

The effects of trans‐spinal direct current (DC) stimulation (tsDCS) on specific neuronal populations are difficult to elucidate, as it affects a variety of neuronal networks. However, facilitatory and depressive effects on neurons processing information from the skin and from muscles can be evaluated separately when weak (0.2–0.3 μA) DC is applied within restricted areas of the rat spinal cord. The effects of such local DC application were recently demonstrated to persist for at least 1 h, and to include changes in the excitability of afferent fibres and their synaptic actions. However, whether these effects require activation of afferent fibres in spinal neuronal pathways during DC application, i.e. whether they are activity‐dependent or activity‐independent, remained an open question. The aim of the present study was to address this question by analysing the effects of local DC application on monosynaptic actions of muscle and skin afferents (extracellular field potentials) and afferent fibre excitability. The results revealed that long‐lasting post‐polarization changes evoked without concomitant activation of afferent fibres replicate changes evoked by stimuli applied during, before and after polarization. The study leads to the conclusion that the reported effects are activity‐independent. As this conclusion applies to the local effects of DC application in at least two spinal pathways and to the effects of both cathodal and anodal polarization, it indicates that some of the more widespread effects of trans‐spinal and trans‐cranial stimulation (both tsDCS and transcranial DC stimulation) may be activity‐independent. The results may therefore contribute to the design of more specific DC applications in clinical practice.

Ischemic block of the forearm abolishes finger movements but not their associated anticipatory postural adjustments

Voluntary movement is known to induce postural perturbations that are counteracted by unconscious anticipatory postural adjustments (APAs). Thus, for every movement, two motor commands are dispatched: a voluntary command recruiting the prime mover and a postural command driving the APAs. These commands are classically thought to be separated; this study investigates whether they could be instead considered as two elements within the same motor program. We analyzed the APAs in biceps brachii, triceps brachii and anterior deltoid that stabilize the arm when briskly flexing the index finger (prime mover flexor digitorum superficialis). APAs and prime mover activation were recorded before, under and after ischemic block of the forearm. Ischemia paralyzed the prime mover, thus suppressing the finger movement and the ensuing postural perturbation. If the two commands had been separated, it would have been expected that after a few failed attempts to flex the index finger, the APAs were suppressed too, being purposeless without postural perturbation. APAs were still present under ischemia even after 60 movement trials. No significant changes were found in APA amplitude in biceps and triceps among different conditions, or in the average APA latency. Inhibitory APA in anterior deltoid was reduced but still present under ischemia. In addition, the pharmacologic block of the sole median nerve produced similar effects. APAs were instead almost abolished when applying a fixation point to the wrist. The observation that APAs remained tailored to the expected perturbation even when that perturbation did not occur supports the idea of a functionally unique motor command driving both the prime mover and the muscles of the APA chain.

A survey of spinal collateral actions of feline ventral spinocerebellar tract neurons.

The aim of this study was to identify spinal target cells of spinocerebellar neurons, in particular the ventral spinocerebellar tract (VSCT) neurons, giving off axon collaterals terminating within the lumbosacral enlargement. Axons of spinocerebellar neurons were stimulated within the cerebellum while searching for most direct synaptic actions on intracellularly recorded hindlimb motoneurons and interneurons. In motoneurons the dominating effects were inhibitory [inhibitory postsynaptic potentials (IPSPs) in 67% and excitatory postsynaptic potentials (EPSPs) in 17% of motoneurons]. Latencies of most IPSPs indicated that they were evoked disynaptically and mutual facilitation between these IPSPs and disynaptic IPSPs evoked by group Ia afferents from antagonist muscles and group Ib and II afferents from synergists indicated that they were relayed by premotor interneurons in reflex pathways from muscle afferents. Monosynaptic EPSPs from the cerebellum were accordingly found in Ia inhibitory interneurons and intermediate zone interneurons with input from group I and II afferents but only oligosynaptic EPSPs in motoneurons. Monosynaptic EPSPs following cerebellar stimulation were also found in some VSCT neurons, indicating coupling between various spinocerebellar neurons. The results are in keeping with the previously demonstrated projections of VSCT neurons to the contralateral ventral horn, showing that VSCT neurons might contribute to motor control at a spinal level. They might thus play a role in modulating spinal activity in advance of any control exerted via the cerebellar loop.

Long-lasting increase in axonal excitability after epidurally applied DC

Effects of direct current (DC) on nerve fibers have primarily been investigated during or just after DC application. However, locally applied cathodal DC was recently demonstrated to increase the excitability of intraspinal preterminal axonal branches for >1 h. The aim of this study was therefore to investigate whether DC evokes a similarly long-lasting increase in the excitability of myelinated axons within the dorsal columns. The excitability of dorsal column fibers stimulated epidurally was monitored by recording compound action potentials in peripheral nerves in acute experiments in deeply anesthetized rats. The results show that 1) cathodal polarization (0.8-1.0 µA) results in a severalfold increase in the number of epidurally activated fibers and 2) the increase in the excitability appears within seconds, 3) lasts for >1 h, and 4) is activity independent, as it does not require fiber stimulation during the polarization. These features demonstrate an unexplored form of plasticity of myelinated fibers and indicate the conditions under which it develops. They also suggest that therapeutic effects of epidural stimulation may be significantly enhanced if it is combined with DC polarization. In particular, by using DC to increase the number of fibers activated by low-intensity epidural stimuli, the low clinical tolerance to higher stimulus intensities might be overcome. The activity independence of long-lasting DC effects would also allow the use of only brief periods of DC polarization preceding epidural stimulation to increase the effect.NEW & NOTEWORTHY The study indicates a new form of plasticity of myelinated fibers. The differences in time course of DC-evoked increases in the excitability of myelinated nerve fibers in the dorsal columns and in preterminal axonal branches suggest that distinct mechanisms are involved in them. The results show that combining epidural stimulation and transspinal DC polarization may dramatically improve their outcome and result in more effective pain control and the return of impaired motor functions.

The Organization and Control of Intra-Limb Anticipatory Postural Adjustments and Their Role in Movement Performance

Anticipatory Postural Adjustments (APAs) are commonly described as unconscious muscular activities aimed to counterbalance the perturbation caused by the primary movement, so as to ensure the whole-body balance, as well as contributing to initiate the displacement of the body center of mass when starting gait or whole-body reaching movements. These activities usually create one or more fixation chains which spread over several muscles of different limbs, and may be thus called inter-limb APAs. However, it has been reported that APAs also precede voluntary movements involving tiny masses, like a flexion/extension of the wrist or even a brisk flexion of the index-finger. In particular, such movements are preceded by an intra-limb APA chain, that involves muscles acting on the proximal joints. Considering the small mass of the moving segments, it is unlikely that the ensuing perturbation could threaten the whole-body balance, so that it is interesting to enquire the physiological role of intra-limb APAs and their organization and control compared to inter-limb APAs. This review is focused on intra-limb APAs and highlights a strict correspondence in their behavior and temporal/spatial organization with respect to inter-limb APAs. Hence it is suggested that both are manifestations of the same phenomenon. Particular emphasis is given to intra-limb APAs preceding index-finger flexion, because their relatively simple biomechanics and the fact that muscular actions were limited to a single arm allowed peculiar investigations, leading to important conclusions. Indeed, such paradigm provided evidence that by granting a proper fixation of those body segments proximal to the moving one APAs are involved in refining movement precision, and also that APAs and prime mover activation are driven by a shared motor command.