Differential Effects of Invasive Anodal Trans-spinal Direct Current Stimulation on Monosynaptic Excitatory Postsynaptic Potentials, Ia Afferents Excitability, and Motoneuron Intrinsic Properties Between Superoxide Dismutase Type-1 Glycine to Alanine Substitution at Position 93 and Wildtype Mice.

In presymptomatic amyotrophic lateral sclerosis (ALS), spinal motoneurons (MNs) have reduced firing patterns and synaptic excitation levels. Preliminary data indicated that in the SOD1 G93A mouse model of ALS, monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in spinal MN by Ia proprioceptive afferent stimulation could be facilitated by trans-spinal direct current stimulation (tsDCS). However, which element of the Ia afferent-MN circuit is affected by tsDCS, and whether tsDCS-induced EPSP facilitation is a general phenomenon or specific to the superoxide dismutase type-1 (SOD1) Glycine to Alanine substitution at position 93 (G93A) mutation, remain to be determined. In this study, we have applied 15-minute tsDCS to the lumbar segments of presymptomatic SOD1 and wildtype (WT) mice and explored its impact on MN passive membrane properties, EPSP amplitude, and Ia afferent activity. While anodal tsDCS induced short-lasting EPSP facilitation in both SOD1 and WT mice, Ia afferent activity and passive membrane properties were altered only in SOD1 mice. Interestingly, EPSP amplitudes of SOD1 mice remained facilitated for at least 1 h after current application, but no long-lasting effect was observed in WT mice. Moreover, anodal tsDCS failed to induce any long-lasting changes in MN passive membrane properties in both SOD1 and WT mice. Conversely, cathodal tsDCS decreased Ia afferent induced EPSP amplitudes only during current application in SOD1 MNs, and no significant effects on Ia afferents excitability were observed. Our findings indicate the high susceptibility of SOD1 MNs to tsDCS and highlight the potential of this neuromodulation technique for the treatment of ALS.

Is there hope that transpinal direct current stimulation corrects motoneuron excitability and provides neuroprotection in amyotrophic lateral sclerosis?

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of largely unknown pathophysiology, characterized by the progressive loss of motoneurons (MNs). We review data showing that in presymptomatic ALS mice, MNs display reduced intrinsic excitability and impaired level of excitatory inputs. The loss of repetitive firing specifically affects the large MNs innervating fast contracting muscle fibers, which are the most vulnerable MNs in ALS. Interventions that aimed at restoring either the intrinsic excitability or the synaptic excitation result in a decrease of disease markers in MNs and delayed neuromuscular junction denervation. We then focus on trans-spinal direct current stimulation (tsDCS), a noninvasive tool, since it modulates the activity of spinal neurons and networks. Effects of tsDCS depend on the polarity of applied current. Recent work shows that anodal tsDCS induces long-lasting enhancement of MN excitability and synaptic excitation of spinal MNs. Moreover, we show preliminary results indicating that anodal tsDCS enhances the excitatory synaptic inputs to MNs in ALS mice. In conclusion, we suggest that chronic application of anodal tsDCS might be useful as a complementary method in the management of ALS patients.

Polarity-dependent adaptations of motoneuron electrophysiological properties after 5-wk transcutaneous spinal direct current stimulation in rats.

Recently, it has been shown that spinal cord polarization considerably modulates motoneuron activity, with certain observed changes in motoneuron membrane and firing properties outlasting the duration of polarization. The purpose of this study was to determine whether repeated sessions of transcutaneous transspinal direct current stimulation (tsDCS) induce adaptive changes in motoneuron properties. In this study, adult male Wistar rats under isoflurane anesthesia were subjected to anodal (n = 6) or cathodal (n = 6) tsDCS (100 µA, 15 min) 5 days per week for 5 wk. Sham control group rats (n = 6) served as a reference. Intracellular recordings from lumbar spinal motoneurons were performed under pentobarbital anesthesia 1 day after the final tsDCS session to analyze membrane and firing properties. Anodal polarization appeared to be effective in evoking significant adaptive changes toward the facilitation of motoneuron firing. When compared with the sham polarization group, these adaptations were expressed by the increased input resistance (P = 0.0077), decreased voltage threshold for spike generation (P = 0.0248) and doublet threshold (P = 0.0311), and increased maximum steady-state firing (SSF) frequency (P = 0.0073), SSF frequency range (P = 0.0075) and slope of the frequency-current relationship (P = 0.0111). However, the effects of cathodal polarization were modest and generally not significant in regard to the sham control. These novel findings support the existing knowledge on alterations in spinal neuronal network excitability in response to polarization and provide the direct evidence of adaptive neuroplasticity of spinal motoneurons in response to chronically applied tsDCS.NEW & NOTEWORTHY Transcutaneous spinal direct current stimulation applied systematically for 5 wk evoked polarity-dependent adaptations in the electrophysiological properties of rat spinal motoneurons. After anodal polarization sessions, motoneurons became more excitable and could evoke higher maximum discharge frequencies during repetitive firing than motoneurons in the sham polarization group. However, no significant adaptive changes of motoneuron properties were observed after repeated cathodal polarization in comparison with the sham control group.

In Vivo Intracellular Recording of Type-Identified Rat Spinal Motoneurons During Trans-Spinal Direct Current Stimulation.

Intracellular recording of spinal motoneurons in vivo provides a "gold standard" for determining the cells' electrophysiological characteristics in the intact spinal network and holds significant advantages relative to classical in vitro or extracellular recording techniques. An advantage of in vivo intracellular recordings is that this method can be performed on adult animals with a fully mature nervous system, and therefore many observed physiological mechanisms can be translated to practical applications. In this methodological paper, we describe this procedure combined with externally applied constant current stimulation, which mimics polarization processes occurring within spinal neuronal networks. Trans-spinal direct current stimulation (tsDCS) is an innovative method increasingly used as a neuromodulatory intervention in rehabilitation after various neurological injuries as well as in sports. The influence of tsDCS on the nervous system remains poorly understood and the physiological mechanisms behind its actions are largely unknown. The application of the tsDCS simultaneously with intracellular recordings enables us to directly observe changes of motoneuron membrane properties and characteristics of rhythmic firing in response to the polarization of the spinal neuronal network, which is crucial for the understanding of tsDCS actions. Moreover, when the presented protocol includes the identification of the motoneuron with respect to an innervated muscle and its function (flexor versus extensor) as well as the physiological type (fast versus slow) it provides an opportunity to selectively investigate the influence of tsDCS on identified components of spinal circuitry, which seem to be differently affected by polarization. The presented procedure focuses on surgical preparation for intracellular recordings and stimulation with an emphasis on the steps which are necessary to achieve preparation stability and reproducibility of results. The details of the methodology of the anodal or cathodal tsDCS application are discussed while paying attention to practical and safety issues.