Continuous Low-frequency Stimulation Research Articles

On-demand low-frequency stimulation for seizure control: efficacy and behavioural implications.

Mesial temporal lobe epilepsy (MTLE), the most common form of focal epilepsy in adults, is often refractory to medication and associated with hippocampal sclerosis. Deep brain stimulation represents an alternative treatment option for drug-resistant patients who are ineligible for resective brain surgery. In clinical practice, closed-loop stimulation at high frequencies is applied to interrupt ongoing seizures, yet has (i) a high incidence of false detections; (ii) the drawback of delayed seizure-suppressive intervention; and (iii) limited success in sclerotic tissue. As an alternative, low-frequency stimulation (LFS) has been explored recently in patients with focal epilepsies. In preclinical epilepsy models, hippocampal LFS successfully prevented seizures when applied continuously. Since it would be advantageous to reduce the stimulation load, we developed a protocol for on-demand LFS. Given the importance of the hippocampus for navigation and memory, we investigated potential consequences of LFS on hippocampal function. To this end, we used the intrahippocampal kainate mouse model, which recapitulates the key features of MTLE, including spontaneous seizure activity and hippocampal sclerosis. Specifically, our online detection algorithm monitored epileptiform activity in hippocampal local field potential recordings and identified short epileptiform bursts preceding focal seizure clusters, triggering hippocampal LFS to stabilize the network state. To probe behavioural performance, we tested the acute influence of LFS on anxiety-like behaviour in the light-dark box test, spatial and non-spatial memory in the object location memory and novel object recognition test, as well as spatial navigation and long-term memory in the Barnes maze. On-demand LFS was almost as effective as continuous LFS in preventing focal seizure clusters but with a significantly lower stimulation load. When we compared the behavioural performance of chronically epileptic mice to healthy controls, we found that both groups were equally mobile, but epileptic mice displayed an increased anxiety level, altered spatial learning strategy and impaired memory performance. Most importantly, with the application of hippocampal LFS before behavioural training and test sessions, we could rule out deleterious effects on cognition and even show an alleviation of deficits in long-term memory recall in chronically epileptic mice. Taken together, our findings may provide a promising alternative to current therapies, overcoming some of their major limitations, and inspire further investigation of LFS for seizure control in focal epilepsy syndromes.

Does low-frequency pelvic nerves stimulation in people with spinal cord injury allow for the formation of electrical pathways responsible for the recovery of walking functions?

Over the last ten years, we have published various manuscripts on the recovery of assisted voluntary walking in people with chronic spinal chord injuries (SCI), following laparoscopic implantation of stimulation electrodes on the pelvic somatic nerves – the LION procedure. Although at the beginning of this research the objective was to allow “robotic” walking by stimulating the muscles, we realized relatively quickly that continuous low frequency stimulation of the pelvic nerves might allow the recovery of voluntary functions of the lower limbs and of the trunk necessary for walking: Seventeen out of a total of twenty-five complete motor chronic SCI-patients (68%) developed enough recovery of supra-spinal control of leg movements, that voluntary walking became feasible, even though a minimal amount of stimulation may be required. All current theories for recovery these voluntary functions below the spinal cord lesion are based on the induced regrowth or reconnection of nerves or at least the recovery of functional anatomical pathways. In this manuscript we formulate the hypothesis that electrical stimulation could be responsible for inducing the formation of “electrical pathways” within the body, which under conditions of electrical stimulation might enable the transport of necessary information from the brain to below the spinal cord lesion allowing voluntary movements of the lower limbs.

Recovery of supraspinal control of leg movement in a chronic complete flaccid paraplegic man after continuous low-frequency pelvic nerve stimulation and FES-assisted training.

More than 30 years ago, functional electrical stimulation (FES) was developed as an orthotic system to be used for rehabilitation for SCI patients. In the present case report, FES-assisted training was combined with continuous low-frequency stimulation of the pelvic somatic nerves in a SCI patient. We report on unexpected findings in a 41-year-old man with chronic complete flaccid paraplegia, since he was 18 years old, who underwent spinal stem cell therapy and a laparoscopic implantation of neuroprosthesis (LION procedure) in the pelvic lumbosacral nerves. The patient had complete flaccid sensomotoric paraplegia T12 as a result of a motor vehicle accident in 1998. In June 2011, he underwent a laparoscopic implantation of stimulation electrodes to the sciatic and femoral nerves for continuous low-frequency electrical stimulation and functional electrical stimulation of the pelvic nerves. Neither intraoperative direct stimulation of the pelvic nerves nor postoperative stimulation induced any sensation or muscle reactions. After 2 years of passive continuous low-frequency stimulation, the patient developed progressive recovery of electrically assisted voluntary motor functions below the lesions: he was first able to extend the right knee and 6 months later, the left. He is currently capable of voluntary weight-bearing standing and walking (with voluntary knee movements) about 50 m with open cuff crutches and drop foot braces. Our findings suggest that continuous low-frequency pelvic nerve stimulation in combination with FES-assisted training might induce changes that affect both the upper and the lower motor neuron and allow supra- and infra-spinal inputs to engage residual spinal and peripheral pathways.

Electrically stimulated sartorius neosphincter: Canine model of activation and skeletal muscle transformation

The sartorius muscle was transposed into the abdominal cavity of six dogs, passed around a Thiry-Vella loop and sutured to itself to form a neosphincter. The muscle was activated by electrical stimulation and on contraction the neosphincter stopped or reduced the flow of saline through the Thiry-Vella loop in all animals until the onset of muscle fatigue. Continuous low frequency stimulation was used to transform the skeletal muscle, and when studied after a mean of 8 weeks of stimulation (range 6-11 weeks) the neosphincter stopped the flow for a significantly longer period of time (P = 0.027). Associated with the improved neosphincter function was a significant decrease in the fusion frequency (P = 0.003) and prolongation of the stimulus-peak tension time as assessed by a strain gauge sutured to the neosphincter muscle (P = 0.002). The changes in the contraction properties of the skeletal muscle suggest that continuous low frequency stimulation transformed the muscle fibres from type 2 to type 1, resulting in improved fatigue resistance and potential for continuous sphincter activity.

Phosphorylation-dependent Translocation of Glycogen Synthase to a Novel Structure during Glycogen Resynthesis

Glycogen metabolism has been the subject of extensive research, but the mechanisms by which it is regulated are still not fully understood. It is well accepted that the rate-limiting enzymes in glycogenesis and glycogenolysis are glycogen synthase (GS) and glycogen phosphorylase (GPh), respectively. Both enzymes are regulated by reversible phosphorylation and by allosteric effectors. However, evidence in the literature indicates that changes in muscle GS and GPh intracellular distribution may constitute a new regulatory mechanism of glycogen metabolism. Already in the 1960s, it was proposed that glycogen was present in dynamic cellular organelles that were termed glycosomas but no such cellular entities have ever been demonstrated. The aim of this study was to characterize muscle GS and GPh intracellular distribution and to identify possible translocation processes of both enzymes. Using in situ stimulation of rabbit tibialis anterior muscle, we show GS and GPh intracellular redistribution at the beginning of glycogen resynthesis after contraction-induced glycogen depletion. We identify a new "player," a new intracellular compartment involved in skeletal muscle glycogen metabolism. They are spherical structures that were not present in basal muscle, and we present evidence that indicate that they are products of actin cytoskeleton remodeling. Furthermore, for the first time, we show a phosphorylation-dependent intracellular distribution of GS. Here, we present evidence of a new regulatory mechanism of skeletal muscle glycogen metabolism based on glycogen enzyme intracellular compartmentalization.

Presynaptic muscarinic receptor mechanisms and submandibular responses to stimulation of the parasympathetic innervation in bursts in rats

Submandibular secretory responses to electrical stimulation of the parasympathetic innervation at variable frequencies were measured in anaesthetized rats. Selective blockade by pirenzepine and by methoctramine occurred at doses (50 and of 300 nmol kg −1, i.v., respectively) that did not inhibit the responses to exogenous acetylcholine. In the presence of methoctramine, the nerve-evoked fluid responses were increased by 100% at 1 Hz independently of the total number of impulses (10–300), suggesting that M2 receptor activation inhibits transmitter release. The magnitude of the increase was inversely related to frequency of stimulation. The protein concentrations in the fluid responses were not significantly affected by methoctramine. Pirenzepine had an inhibitory effect on the fluid secretory responses, which was dependent of frequency, as well as of number of impulses, suggesting that M1 receptor activation facilitates transmitter release. At 10 Hz given intermittently (for 1 s at 10-s intervals), pirenzepine reduced the fluid response by 25%. The protein release was substantially and significantly reduced by pirenzepine independent of frequency but only during long periods of stimulation (300 impulses). It is concluded that muscarinic M1 receptor activation normally has a facilitatory effect on transmitter release, and that the facilitation occurs during short, intense stimulation. Muscarinic M1 receptors are, however, likely to regulate protein secretion by other mechanisms. Muscarinic M2 receptors, on the other hand, normally inhibit cholinergic transmission at low frequencies. Similar to findings in the alimentary tract of several species, stimulation in bursts at high frequencies is a more efficient stimulation pattern than continuous low frequency stimulation. This pattern of stimulation thus takes advantage of transient facilitation and avoids the inhibition at less intense neuronal activity.

Inactivation of sarcoplasmic reticulum Ca(2+)-atpase in low-frequency stimulated rat muscle.

Continuous low-frequency stimulation (CLFS) by implanted electrodes for 12-24 h led to a significant (approximately 30%) decrease in the activity of sarcoplasmic reticulum Ca(2+)-ATPase in fast-twitch extensor digitorum longus (EDL) and tibialis anterior (TA) muscles of intact rats. The decline in catalytic activity after 24 h of CLFS was accompanied by an approximately twofold increase in dinitrophenylhydrazine-reactive carbonyl groups of the enzyme. It also correlated with an immunochemically determined 30% decrease in Ca2(+)-ATPase protein. Recovery studies after 12 h of CLFS revealed a relatively slow (48-72 h) re-establishment of normal catalytic activity. These findings suggest that the 30% decline of Ca(2+)-ATPase activity in low-frequency stimulated rat muscle led to an irreversible modification by protein oxidation. The decrease in Ca(2+)-ATPase protein most likely resulted from the degradation of inactive Ca(2+)-ATPase molecules. The relatively slow recovery of Ca(2+)-ATPase activity suggests that de novo synthesis of the enzyme may be necessary to re-attain normal activity.

Effect of low and high frequency thalamic stimulation on sleep in patients with Parkinson's disease and essential tremor.

Continuous high frequency stimulation of the ventral intermediate nucleus of the thalamus (Vim), delivered through surgically implanted quadripolar electrodes, alleviates tremor in Parkinson's disease (PD) and essential tremor (ET). The Vim is adjacent to the thalamic reticular nuclei, where sleep spindles originate according to animal models. In order to determine whether Vim stimulation affects sleep spindles, six patients (4 PD, 2 ET), aged 60-69 years, were recorded on a control night and a stimulation night (130 Hz, 2-3 V; right stimulation in five patients and bilateral stimulation in one patient). Stimulation did not modify sleep quality or architecture. Sleep spindles were present and symmetrical in five out of six patients under stimulation. However, in one patient with a sustained 'thalamotomy-like effect' that abolished tremor, spindles were asymmetrical even without stimulation. In each patient, spindle density was similar on both nights (mean+/- SEM: 2.25+/-0. 61 spindles per min of stage 2 sleep vs. 1.84+/-0.31). In an attempt to promote sleep two different patterns of stimulation were applied in the region of ventrooralis posterior and reticularis nuclei in five patients in the awake state. Continuous low frequency stimulation (5 Hz, 0.1 V), and repeated trains of 15 Hz for 1 s every 15 s mimicking the pattern of physiological spindles, each failed to induce sleep or cortical synchronization. We conclude that Vim stimulation, unlike thalamotomy, selectively reduces tremor without altering sleep or sleep spindles. Our results also suggest that low frequency stimulation applied in the region of the reticular nuclei does not induce sleep.

Nervous control of tracheal blood flow in the cat measured by the laser Doppler technique.

The tracheal blood flow as determined by the laser Doppler technique was continuously monitored in anaesthetized cats. Electrical stimulation of the right superior laryngeal nerve caused an atropine-resistant increase in blood flow of the upper trachea. Unilateral vagal nerve stimulation at the cervical level in the presence of atropine induced a frequency-dependent increase in blood flow of the lower trachea. Intermittent stimulation with bursts of impulses at a high frequency resulted in a considerably larger blood flow increase than a continuous low frequency stimulation giving the same total number of impulses. The ganglionic blocking agent chlorisondamine abolished most of the vagally induced increase in tracheal blood flow when using low threshold parameters (2 V, 0.2 ms) presumably activating preganglionic nerves. High threshold stimulation (10 V, 5 ms) however, still resulted in an increased blood flow suggesting antidromic activation of sensory C fibres. Local mechanical irritation or chemical irritation by capsaicin also increased tracheal blood flow. Furthermore, local application of calcitonin gene-related peptide on to the mucosa caused a slowly developing, long-lasting increase in blood flow. Electrical stimulation of the cervical sympathetic trunk and local application of adrenaline reduced tracheal blood flow. In conclusion, vagal nerve stimulation induces an atropine-resistant increase in tracheal blood flow probably mainly by activating preganglionic parasympathetic nerves and possibly also by antidromic stimulation of C-fibre afferents.

The effect of long-term high-frequency stimulation on capillary density and fibre types in rabbit fast muscles.

Rabbit fast muscles (tibialis anterior, t.a.; extensor digitorum longus, e.d.l.; and peroneal muscles) were stimulated for up to 28 days by electrodes implanted in the vicinity of the lateral popliteal (peroneal) nerve for 8 h/day, using either intermittent high-frequency (three trains at 40 Hz/min, each 5 s duration), or continuous stimulation at 10 Hz. This did not result in muscle hypertrophy even after 28 days. Capillary density (number of capillaries/mm2) was increased in e.d.l. from 251 +/- 3 to 366 +/- 6 after 14 days of stimulation and from 251 +/- 3 to 514 +/- 13 after 28 days of stimulation at 40 Hz. In t.a., capillary density increased from 373 +/- 5 to 583 +/- 10 after 14 days of stimulation at 40 Hz. The capillary/fibre ratio increased in e.d.l. from 1.25 +/- 0.02 to 1.86 +/- 0.04 at 14 days and to 2.07 +/- 0.06 at 28 days. In t.a., capillary/fibre ratio increased from 1.40 +/- 0.03 to 1.83 +/- 0.05 at 14 days. All these changes were significant (P less than 0.0005). Analysis of capillary density, capillary/fibre ratio, fibre areas and proportion of different fibre types in muscles stimulated for shorter periods showed no changes in capillary density, capillary/fibre ratio or fibre areas in e.d.l. or t.a. stimulated for 4 days; there was a decrease in the proportion of fast glycolytic fibres from 42 to 32% (P less than 0.0025) and increase in fast oxidative from 37.6 to 41.2% in e.d.l. Muscles stimulated for 7 days showed increases in capillary density and capillary/fibre ratio in fast predominantly glycolytic fibres in e.d.l., and a decrease in capillary density in fast and slow oxidative fibres in t.a. This was partly due to the increase in fibre areas in these groups (capillary/fibre ratio in t.a. was not significantly changed). No changes were observed in fibre areas in e.d.l. Stimulation at 10 Hz produced increase in capillary/fibre ratio in the vicinity of glycolytic fibres after only 4 days. High-frequency intermittent stimulation leads to a massive capillary growth which starts first in the muscle with a higher proportion of glycolytic fibres (e.d.l.), has a later onset than continuous low-frequency stimulation, and may be due to a combination of high blood flow and metabolic factors.

Inhibition and Rhythmic Activity of the Circular Muscles of Calliactis Parasitica (Couch)

ABSTRACT The responses to electrical stimulation of isolated rings of the column and pedal disk of Calliactis are described. Such rings make slow spontaneous contractions which are frequently rhythmical, the interval between contractions normally being 7-20 min. Continuous low-frequency stimulation inhibits spontaneous activity of rings from the pedal disk and also of fresh rings from more adoral regions of the column. Older rings from the mid-column respond to such stimulation by a tetanic contraction. The latency of response to electrical stimulation of pedal rings is of the order of 120 sec. This latency is not affected by stimulation frequency but is prolonged by increase in the number of shocks applied. Stimulation of a pedal ring at the onset of a contraction prevents the further development of this contraction, while stimulation as a contraction reaches its maximum is followed by more rapid relaxation than in unstimulated controls. Mid-column rings when freshly prepared show a latency of the order of 120 sec. As the preparation ages, a double response to stimulation appears ; the first response has a latency of about 30-40 sec. and presently becomes the only type of response shown. If two sets of stimuli are applied to a mid-column ring, the magnitude of response to the second set increases as the time between stimulations increases. With long intervals an almost total contraction is obtained in response to a single shock. The effect of intercalated stimuli upon the rhythm of spontaneous activity is studied. The effect is very variable and it is suggested that this is the result of electrical stimulation having both an excitatory and an inhibitory effect. The very long latent periods characteristic of pedal rings and the rhythmic activity of these preparations are interpreted as interactions of excitation and inhibition.