Selective Stimulation Research Articles

Magnetic Stimulation of Gigantocellular Reticular Nucleus with Iron Oxide Nanoparticles Combined Treadmill Training Enhanced Locomotor Recovery by Reorganizing Cortico-Reticulo-Spinal Circuit.

Gigantocellular reticular nucleus (GRNs) executes a vital role in locomotor recovery after spinal cord injury. However, due to its unique anatomical location deep within the brainstem, intervening in GRNs for spinal cord injury research is challenging. To address this problem, this study adopted an extracorporeal magnetic stimulation system to observe the effects of selective magnetic stimulation of GRNs with iron oxide nanoparticles combined treadmill training on locomotor recovery after spinal cord injury, and explored the possible mechanisms. Superparamagnetic iron oxide (SPIO) nanoparticles were stereotactically injected into bilateral GRNs of mice with moderate T10 spinal cord contusion. Eight-week selective magnetic stimulation produced by extracorporeal magnetic stimulation system (MSS) combined with treadmill training was adopted for the animals from one week after surgery. Locomotor function of mice was evaluated by the Basso Mouse Scale, Grid-walking test and Treadscan analysis. Brain MRI, anterograde virus tracer and immunofluorescence staining were applied to observe the tissue compatibility of SPIO in GRNs, trace GRNs' projections and evaluate neurotransmitters' expression in spinal cord respectively. Motor-evoked potentials and H reflex were collected for assessing the integrity of cortical spinal tract and the excitation of motor neurons in anterior horn. (1) SPIO persisted in GRNs for a minimum of 24 weeks without inducing apoptosis of GRN cells, and degraded slowly over time. (2) MSS-enabled treadmill training dramatically improved locomotor performances of injured mice, and promoted cortico-reticulo-spinal circuit reorganization. (3) MSS-enabled treadmill training took superimposed roles through both activating GRNs to drive more projections of GRNs across lesion site and rebalancing neurotransmitters' expression in anterior horn of lumbar spinal cord. These results indicate that selective MSS intervention of GRNs potentially serves as an innovative strategy to promote more spared fibers of GRNs across lesion site and rebalance neurotransmitters' expression after spinal cord injury, paving the way for the structural remodeling of neural systems collaborating with exercise training, thus ultimately contributing to the reconstruction of cortico-reticulo-spinal circuit.

Stretchable Tissue-Like Gold Nanowire Composites with Long-Term Stability for Neural Interfaces.

Soft and stretchable nanocomposites can match the mechanical properties of neural tissue, therebyminimizing foreign body reactions to provide optimal stimulation and recording specificity. Soft materials for neural interfaces should simultaneously fulfill a wide range of requirements, including low Young's modulus (<<1MPa), stretchability (≥30%), high conductivity (>> 1000 Scm-1), biocompatibility, and chronic stability (>> 1 year). Current nanocomposites do not fulfill the above requirements, in particular not the combination of softness and high conductivity. Here, this challenge is addressed by developing a scalable and robust synthesis route based on polymeric reducing agents for smooth, high-aspect ratio gold nanowires (AuNWs) of controllable dimensions with excellent biocompatibility. AuNW-silicone composites show outstanding performance with nerve-like softness (250kPa), high conductivity (16000 Scm-1), and reversible stretchability. Soft multielectrode cuffs based on the composite achieve selective functional stimulation, recordings of sensory stimuli in rat sciatic nerves, and show an accelerated lifetime stability of >3 years. The scalable synthesis method provides a chemically stable alternative to the widely used AgNWs, thereby enabling new applications within electronics, biomedical devices, and electrochemistry.

3D electronic implants in subretinal space: Long-term follow-up in rodents

Clinical results with photovoltaic subretinal prosthesis (PRIMA) demonstrated restoration of sight via electrical stimulation of the interneurons in degenerated retina, with resolution matching the 100 μm pixel size. Since scaling the pixels below 75 μm in the current bipolar planar geometry will significantly limit the penetration depth of the electric field and increase stimulation threshold, we explore the possibility of using smaller pixels based on a novel 3-dimensional honeycomb-shaped design. We assessed the long-term biocompatibility and stability of these arrays in rats by investigating the anatomical integration of the retina with flat and 3D implants and response to electrical stimulation over lifetime – up to 32–36 weeks post-implantation in aged rats. With both flat and 3D implants, signals elicited in the visual cortex decreased after the day of implantation by more than 3-fold, and gradually recovered over the next 12–16 weeks. With 25 μm high honeycomb walls, the majority of bipolar cells migrate into the wells, while amacrine and ganglion cells remain above the cavities, which is essential for selective network-mediated stimulation of the retina. Retinal thickness and full-field stimulation threshold with 40 μm-wide honeycomb pixels were comparable to those with planar devices – 0.05 mW/mm2 with 10 ms pulses. However, fewer cells from the inner nuclear layer migrated into the 20 μm-wide wells, and stimulation threshold increased over 12–16 weeks, before stabilizing at about 0.08 mW/mm2. Such threshold is still significantly lower than 1.8 mW/mm2 with a previous design of flat bipolar pixels, confirming the promise of the 3D honeycomb-based approach to high resolution subretinal prosthesis.

Neuron type-specific optogenetic stimulation for differential stroke recovery in chronic capsular infarct

Cortical neuromodulation (CNM) is widely used to promote recovery after stroke. Despite the beneficial results of CNM, the roles played by different neuron types in the effects of current CNM techniques are unable to be differentiated. Our aim was to use selective optogenetic cortical stimulation to explore how different subpopulations of neuronal cells contribute to poststroke recovery. We transduced the sensory-parietal cortex (SPC) of rats with CamKII-ChR2 (pyramidal neurons), PV-ChR2 (parvalbumin-expressing inhibitory neurons), or hSyn-ChR2 (pan-neuronal population) before inducing photothrombotic capsular infarct lesions. We found that selective stimulation of inhibitory neurons resulted in significantly greater motor recovery than stimulation of excitatory neurons or the pan-neuronal population. Furthermore, 2-deoxy-2-[18F] fluoro-D-glucose microPET (FDG-microPET) imaging revealed a significant reduction in cortical diaschisis and activation of the corticostriatal neural circuit, which were correlated with behavioral recovery in the PV-ChR2 group. The spatial pattern of brain-derived neurotrophic factor (BDNF) expression was evident in the stimulated cortex and underlying cortico-subcortical circuit. Our results indicate that the plasticity of inhibitory neurons is crucial for functional recovery after capsular infarct. Modifying CNM parameters to potentiate the stimulation of inhibitory neurons could improve poststroke outcomes.

A Cell-Type Selective Stimulation and Recording System for Retinal Ganglion Cells.

Future retinal implants will require a stimulation selectivity between different sub-types of Retinal Ganglion Cells (RGCs) to evoke natural perceptions rather than phosphenes in patients. To achieve this, a cell-type specific stimulation pipeline is required that identifies target RGC sub-types from recorded input images and extracts the specific stimulation parameters to activate this cell-type selectively. Promising biological experiments showed that ON-/OFF- sustained/transient RGCs could be selectively activated by modulating repetition rate and amplitude of an electrical stimulation current in the kilohertz range. This research presents a 42 channel current controlled stimulation and recording system on chip (SoC) with parameter input from a real time target RGC selection algorithm. The SoC is able to stimulate retinal tissue with sinusoidal frequencies higher than 1 kHz at amplitudes of up to 200 μA at a supply voltage of 1.8 V. It also includes tunable recording units with an integrated action potential detection pipeline that are able to amplify signals between 1 Hz and 50 kHz. The required area of one stimulator is 0.0071 mm2, while one recording unit consumes an area of 0.0092 mm2. The application of sinusoidal stimulation currents in the kilohertz range towards retinal tissue leads to a suppressive response of only certain RGC sub-types that has not been oberved before, using electrical stimulation. Because this response is very similar to the natural light response of RGCs, this stimulation approach can lead to a more genuine visual perception for patients using retinal implants.

TRPV1 nociceptors are required to optimize antigen-specific primary antibody responses to novel antigens

BackgroundKey to the advancement of the field of bioelectronic medicine is the identification of novel pathways of neural regulation of immune function. Sensory neurons (termed nociceptors) recognize harmful stimuli and initiate a protective response by eliciting pain and defensive behavior. Nociceptors also interact with immune cells to regulate host defense and inflammatory responses. However, it is still unclear whether nociceptors participate in regulating primary IgG antibody responses to novel antigens.MethodsTo understand the role of transient receptor potential vanilloid 1 (TRPV1)-expressing neurons in IgG responses, we generated TRPV1-Cre/Rosa-ChannelRhodopsin2 mice for precise optogenetic activation of TRPV1 + neurons and TRPV1-Cre/Lox-diphtheria toxin A mice for targeted ablation of TRPV1-expressing neurons. Antigen-specific antibody responses were longitudinally monitored for 28 days.ResultsHere we show that TRPV1 expressing neurons are required to develop an antigen-specific immune response. We demonstrate that selective optogenetic stimulation of TRPV1+ nociceptors during immunization significantly enhances primary IgG antibody responses to novel antigens. Further, mice rendered deficient in TRPV1- expressing nociceptors fail to develop primary IgG antibody responses to keyhole limpet hemocyanin or haptenated antigen.ConclusionThis functional and genetic evidence indicates a critical role for nociceptor TRPV1 in antigen-specific primary antibody responses to novel antigens. These results also support consideration of potential therapeutic manipulation of nociceptor pathways using bioelectronic devices to enhance immune responses to foreign antigens.

Pillar electrodes embedded in the skeletal muscle tissue for selective stimulation of biohybrid actuators with increased contractile distance.

Electrodes are crucial for controlling the movements of biohybrid robots, but their external placement outside muscle tissue often leads to inefficient and non-selective stimulation of nearby biohybrid actuators. To address this, we propose embedding pillar electrodes within the skeletal muscle tissue, resulting in enhanced contraction of the target muscle without affecting the neighbor tissue with a 4 mm distance. We use finite element method simulations to establish a selectivity model, correlating the VIE(volume integration of electric field intensity within muscle tissue) with actual contractile distances under different amplitudes of electrical pulses. The simulated selective index closely aligns with experimental results, showing the potential of pillar electrodes for effective and selective biohybrid actuator stimulation. In experiments, we validated that the contractile distance and selectivity achieved with these pillar electrodes exceed conventional Au rod electrodes. This innovation has promising implications for building biohybrid robots with densely arranged muscle tissue, ultimately achieving more human-like movements. Additionally, our selectivity model offers valuable predictive tools for assessing electrical stimulation effects with different electrode designs.

The influence of subclavian ansae stimulation on cardiac hemodynamics and electrophysiology in atrial fibrillation- exploring a novel target for neuromodulation in atrial fibrillation (SAS-AF study)

Abstract Background Sympathetic nervous system activation plays a significant role in arrhythmia development and maintenance.Neuromodulation techniques such as stellate ganglion block and sympathectomy, are limited by off-target side effects and complex targeting.The Subclavian Ansae (SA,a nerve cord encircling the subclavian artery) may be an intravascular site allowing specific targeting of the cardiac afferent sympathetic system.Preclinical studies have revealed increased sympathetic tone upon selective SA stimulation. Objective We sought to define for the first time in Humans the functional anatomy and physiology of Subclavian Ansae via a percutaneous approach. Methods A prior cadaveric study defined the anatomical course of the subclavian ansae, encircling the subclavian arteries and connecting the middle cervical ganglia to inferior cervical ganglia/stellate.Patients undergoing catheter ablation for paroxysmal atrial fibrillation(pAF) under general anesthesia were prospectively recruited for the study. Subclavian Ansae stimulation (SAS) was performed on the left and/or the right (L/SAS ± R/SAS) within the subclavian artery using the TactiCath Ablation Catheter introduced via a femoral arterial sheath. Stimulation parameters included up to 70 V output, 10 Hz frequency, and pulse width of 2-4ms for 20-30s. Test pulses ensured no cardiac capture during stimulation. Participants discontinued antiarrhythmic agents at least 4-5 half-lives prior to the procedure. Invasive arterial blood pressure(BP), heart rate (HR), and electrophysiological parameters were recorded in response to selective stimulation. An increase of ten percent or more in either arterial blood pressure(BP) or heart rate(HR) from the initial baseline measurements was deemed a positive response. Results 15 patients (62±11 years, 9 males, pAF duration 51±24 months) underwent the simulation protocol prior to their clinical AF ablation procedure. A positive hemodynamic response was observed in nine patients, and among them, arrhythmia was inducible in 3 patients (atrial fibrillation in 2 patients and repeated runs of atrial ectopy in 1 patient) in response to SAS. Mean sinus cycle length(CL) decreased(chronotropic) after stimulation,particularly with a significant change observed in response to R/SAS. Additionally,L/SAS led to a notable increase in mean systolic (SBP) and diastolic(DBP) blood pressure (inotropic). While the mean right atrial effective refractory period (ERP) did not significantly decrease after stimulation, there was a demonstrable difference in interatrial conduction time(dromotropy). Six patients, including the initial 3 subjected to low-energy stimulation (up to 12V), did not exhibit an adrenergic response. Conclusion This study represents the first successful application of selective SAS in humans. A heterogeneity in electrophysiological modulation by SAS exists, including laterality.The SA is a potential important target for targeted autonomic neuromodulation therapy

Selective intrafascicular stimulation of myelinated and unmyelinated nerve fibers through a longitudinal electrode: A computational study

Carbon nanotube (CNT) fiber electrodes have demonstrated exceptional spatial selectivity and sustained reliability in the context of intrafascicular electrical stimulation, as evidenced through rigorous animal experimentation. A significant presence of unmyelinated C fibers, known to induce uncomfortable somatosensory experiences, exists within peripheral nerves. This presence poses a considerable challenge to the excitation of myelinated Aβ fibers, which are crucial for tactile sensation. To achieve nuanced tactile sensory feedback utilizing CNT fiber electrodes, the selective stimulation of Aβ sensory afferents emerges as a critical factor.In confronting this challenge, the present investigation sought to refine and apply a rat sciatic-nerve model leveraging the capabilities of the COMSOL-NEURON framework. This approach enables a systematic evaluation of the influence exerted by stimulation parameters and electrode geometry on the activation dynamics of both myelinated Aβ and unmyelinated C fibers. The findings advocate for the utilization of current pulses featuring a pulse width of 600 μs, alongside the deployment of CNT fibers characterized by a diminutive diameter of 10 μm, with an exclusively exposed cross-sectional area, to facilitate reduced activation current thresholds. Comparative analysis under monopolar and bipolar electrical stimulation conditions revealed proximate activation thresholds, albeit with bipolar stimulation exhibiting superior fiber selectivity relative to its monopolar counterpart. Concerning pulse waveform characteristics, the adoption of an anodic-first biphasic stimulation modality is favored, taking into account the dual criteria of activation threshold and fiber selectivity optimization. Consequently, this investigation furnishes an efficacious stimulation paradigm for the selective activation of touch-related nerve fibers, alongside provisioning a comprehensive theoretical foundation for the realization of natural tactile feedback within the domain of prosthetic hand applications.

The laterodorsal tegmental nucleus improves breathing and sleep in a Parkinson’s disease animal model

Parkinson's disease (PD) is a neurological disorder resulting from the degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNpc). PD is characterized by classical and non-classical symptoms including respiratory instability and sleep disturbances. An important region for the control of sleep and possibly respiration is the laterodorsal tegmental nucleus (LDTg), a nucleus composed of cholinergic, glutamatergic, and GABAergic neurons. We hypothesize that stimulation of cholinergic neurons in the LDTg can lead to improvements in sleep and respiratory disturbances in a murine model of PD. Adult mice (CEUA: 6641200919 and SCRI: 18819) that express the fluorescent green protein in cholinergic cells (Chat-cre, ai6) received bilateral injections of AAV-hM3Dq-mCherry or AAV-mCherry vectors into LDTg. After ten days, the animals received nigrostriatal injections of vehicle or 6-hydroxydopamine (6-OHDA, 10 μg/μl). Eight days later, we implanted EEG and EMG electrodes. In 6-OHDA-injected mice, there was a 79% and 29% reduction in the number of dopaminergic neurons in SNpc and cholinergic neurons in LDTg, respectively. This reduction was associated with an increase in the number of apneas and a 17% and 26% reduction in respiratory frequency during non-REM (NREMs) and REM (REMs) sleep, respectively. In the same animals, there was also a decrease in wakefulness time, and an increase in total sleep time. Selective pharmacogenetic stimulation of ChAT+ neurons of the LDTg rescued the respiratory frequency during REMs and reduced the number of apneas both in NREMs and REMs in 6-OHDA mice. These results demonstrate that in this PD model the LDTg plays a significant role in respiratory changes associated with sleep regulation. FAPESP, CAPES, CNPq and NIH. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A Morphologically Distinct Subclass of Somatostatin-expressing Interneurons in the Somatosensory Cortex are Strongly Recruited by Input from the Motor Cortex

The motor cortex (M1) and somatosensory cortex (S1) are strongly interconnected regions, and their interactions are essential for sensory perception and motor execution. However, the mechanisms by which M1 activity modulates sensory processing within S1 are poorly understood. In particular, M1 recruitment of distinct GABAergic inhibitory cells could impact S1 responsiveness. Preliminary work in our lab identified a subpopulation of somatostatin (SOM) expressing inhibitory interneurons in layer 6 (L6) of S1 that are strongly recruited by M1 input. We hypothesize these M1-responding L6 SOM cells are an electrophysiological and morphologically distinct subclass that express the enzyme neuronal nitric oxide synthase (nNOS). To stimulate the M1 to S1 pathway and test this hypothesis, we injected adeno-associated virus (AAV) encoding the light-sensitive cation channel, channelrhodopsin-2 (ChR2), in M1 of postnatal day 21 (+/-1) mice in vivo. After three weeks of expression, we prepared acute coronal brain slices for targeted loose-patch recordings and selective optical stimulation of M1 terminals. We found two groups of L6 SOM cells based on their spiking behavior during photostimulation: responsive (15%) and non-responsive (85%). Whole-cell recordings and neurobiotin injections into responsive and non-responsive SOM cells revealed robust electrophysiological and morphological differences. Initial anatomical reconstructions indicate that the non-responsive SOM cells (n=12) exhibit both Martinotti (with an L1 axonal projection) and non-Martinotti (no L1 axonal projection) morphologies, whereas the responsive SOM cells had axonal arborizations projecting toward the underlying white matter (n=10). The responsive SOM cells had quasi-fast-spiking electrophysiological properties (n=10) and were negative for nNOS (n=9), whereas the non-responsive SOM cells exhibited non-adaptive spiking behavior. Importantly, responsive SOM cells were negative for parvalbumin (PV), a marker for a distinct class of fast-spiking interneurons in the cortex, indicating these cells were not mislabeled PV cells due to the known off-target recombination in the SOM-IRES-Cre mouse line (n=4). In summary, our data show that input from M1 strongly recruits a previously unknown SOM-expressing interneuron in lower L6 with distinct morphological and electrophysiological features that could influence sensory responsiveness in S1. Future studies will focus on the mechanisms of this selective activation and identifying the postsynaptic targets of these cells to further our understanding of this sensorimotor circuit. NIH R01NS117636 (SRC); APS SURF (MKS). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Characterization, number, and spatial organization of nerve fibers in the human cervical vagus nerve and its superior cardiac branch

BackgroundElectrical stimulation of the vagus nerve (VN) is a therapy for epilepsy, obesity, depression, and heart diseases. However, whole nerve stimulation leads to side effects. We examined the neuroanatomy of the mid-cervical segment of the human VN and its superior cardiac branch to gain insight into the side effects of VN stimulation and aid in developing targeted stimulation strategies. MethodsNerve specimens were harvested from eight human body donors, then subjected to immunofluorescence and semiautomated quantification to determine the signature, quantity, and spatial distribution of different axonal categories. ResultsThe right and left cervical VN (cVN) contained a total of 25,489 ± 2781 and 23,286 ± 3164 fibers, respectively. Two-thirds of the fibers were unmyelinated and one-third were myelinated. About three-quarters of the fibers in the right and left cVN were sensory (73.9 ± 7.5 % versus 72.4 ± 5.6 %), while 13.2 ± 1.8 % versus 13.3 ± 3.0 % were special visceromotor and parasympathetic, and 13 ± 5.9 % versus 14.3 ± 4.0 % were sympathetic. Special visceromotor and parasympathetic fibers formed clusters. The superior cardiac branches comprised parasympathetic, vagal sensory, and sympathetic fibers with the left cardiac branch containing more sympathetic fibers than the right (62.7 ± 5.4 % versus 19.8 ± 13.3 %), and 50 % of the left branch contained sensory and sympathetic fibers only. ConclusionThe study indicates that selective stimulation of vagal sensory and motor fibers is possible. However, it also highlights the potential risk of activating sympathetic fibers in the superior cardiac branch, especially on the left side.

Interleukin-15 is a hair follicle immune privilege guardian

The autoimmunity-promoting cytokine, Interleukin-15 (IL-15), is often claimed to be a key pathogenic cytokine in alopecia areata (AA). Yet, rhIL-15 promotes human hair follicle (HF) growth ex vivo. We have asked whether the expression of IL-15 and its receptor (IL-15R) isoforms is altered in human AA and how IL-15 impacts on human HF immune privilege (HF-IP) in the presence/absence of interferon-γ (IFNγ), the well-documented key AA-pathogenic cytokine, as well as on hair regrowth after experimental AA induction in vivo. Quantitative immunohistomorphometry showed the number of perifollicular IL-15+ T cells in AA skin biopsies to be significantly increased compared to healthy control skin, while IL-15, IL-15Rα, and IL-15Rγ protein expression within the hair bulb were significantly down-regulated in AA HFs. In organ-cultured human scalp HFs, rhIL-15 significantly reduced hair bulb expression of MICA, the key “danger” signal in AA pathogenesis, and increased production of the HF-IP guardian, α-MSH. Crucially, ex vivo, rhIL-15 prevented IFNγ-induced HF-IP collapse, restored a collapsed HF-IP by IL-15Rα-dependent signaling (as documented by IL-15Rα−silencing), and protected AA-preventive immunoinhibitory iNKT10 cells from IFNγ-induced apoptosis. rhIL-15 even promoted hair regrowth after experimental AA induction in human scalp skin xenotransplants on SCID/beige mice in vivo. Our data introduce IL-15 as a novel, functionally important HF-IP guardian whose signaling is constitutively defective in scalp HFs of AA patients. Our data suggest that selective stimulation of intrafollicular IL-15Rα signaling could become a novel therapeutic approach in AA management, while blocking it pharmacologically may hinder both HF-IP restoration and hair re-growth and may thus make HFs more vulnerable to AA relapse.

Avoidance of axonal stimulation with sinusoidal epiretinal stimulation

Objective. Neuromodulation, particularly electrical stimulation, necessitates high spatial resolution to achieve artificial vision with high acuity. In epiretinal implants, this is hindered by the undesired activation of distal axons. Here, we investigate focal and axonal activation of retinal ganglion cells (RGCs) in epiretinal configuration for different sinusoidal stimulation frequencies. Approach. RGC responses to epiretinal sinusoidal stimulation at frequencies between 40 and 100 Hz were tested in ex-vivo photoreceptor degenerated (rd10) isolated retinae. Experiments were conducted using a high-density CMOS-based microelectrode array, which allows to localize RGC cell bodies and axons at high spatial resolution. Main results. We report current and charge density thresholds for focal and distal axon activation at stimulation frequencies of 40, 60, 80, and 100 Hz for an electrode size with an effective area of 0.01 mm2. Activation of distal axons is avoided up to a stimulation amplitude of 0.23 µA (corresponding to 17.3 µC cm−2) at 40 Hz and up to a stimulation amplitude of 0.28 µA (14.8 µC cm−2) at 60 Hz. The threshold ratio between focal and axonal activation increases from 1.1 for 100 Hz up to 1.6 for 60 Hz, while at 40 Hz stimulation frequency, almost no axonal responses were detected in the tested intensity range. With the use of synaptic blockers, we demonstrate the underlying direct activation mechanism of the ganglion cells. Finally, using high-resolution electrical imaging and label-free electrophysiological axon tracking, we demonstrate the extent of activation in axon bundles. Significance. Our results can be exploited to define a spatially selective stimulation strategy avoiding axonal activation in future retinal implants, thereby solving one of the major limitations of artificial vision. The results may be extended to other fields of neuroprosthetics to achieve selective focal electrical stimulation.

Prebiotic effects of dietary xylooligosaccharides on fish gut microbiota, growth, and immunological parameters – A review

Abstract Xylooligosaccharides (XOSs) are being explored as prebiotics in fish diets; however, their effects and modes of action have not been well evaluated. Reports have shown that dietary XOS has the potential to improve the proliferation of beneficial gut microbes, and their metabolites, and enhance disease resistance in several fish species. In contrast, other studies report no substantial changes in immune and growth parameters compared to control groups. Like all prebiotics, the mode of action of XOS is based on their selective stimulation of beneficial gut microbiota, which will outcompete and prevent pathogen proliferation in the gut, and produce metabolites that modulate host immune responses. The reports of improved growth performance of XOS fed fish may be due to improved intestinal microbiome, enhanced glycolysis activity and elevated gastrointestinal enzymatic activities. Dietary XOSs have different effects on fish performance depending on the fish species and the structure of XOSs (degree of XOS polymerization and substitution). Nevertheless, further research is essential to determine the optimal dosage, degree of polymerization, and substitution levels required to improve each fish species’ gut health and growth performance. This review highlights the prebiotic effects of XOSs, their mechanism of action, and knowledge gaps.

New insights into the characterization of the mechanism of action of hyoscine butylbromide in the human colon ex vivo

IntroductionHyoscine butylbromide (HBB) is one of the most used antispasmodics in clinical practice. Recent translational consensus has demonstrated a similarity between human colonic motor patterns studied ex vivo and in vivo, suggesting ex vivo can predict in vivo results. It is unclear whether the mechanism of action of antispasmodics can predict different use in clinical practice. The aim of the present study is to bridge this gap dissecting HBB's role in excitatory and inhibitory neural pathways. Methods: 309 colon samples from 48 patients were studied in muscle bath experiments. HBB was tested on: 1-spontaneous phasic contractions (SPCs); 2-carbachol-induced contractility; electrical field stimulation (EFS)-induced selective stimulation of 3-excitatory and 4-inhibitory pathways and 5- SPCs and EFS-induced contractions enhanced by neostigmine. Atropine, AF-DX116 (M2 blocker) and DAU-5884 (M3 blocker) were used as comparators. ResultsIn the presence of tetrodotoxin (TTX), HBB and atropine 1 μM reduced SPCs. HBB and atropine concentration-dependently reduced carbachol- and EFS-induced contractions. Inhibitory effects of DAU-5884 on EFS-induced contractions were more potent than of AF-DX116. HBB did not affect the off-response associated to neural inhibitory responses. Neostigmine enhanced both SPCs and EFS-induced contractions. In the presence of TTX and ω-conotoxin (GVIA), neostigmine still enhanced SPCs. Addition of HBB and atropine reduced these responses. ConclusionsThis study demonstrates that HBB inhibits neural cholinergic contractions associated to muscarinic (mainly M3) receptors. HBB has a potential role in reducing colonic spasm induced by the release of acetylcholine from enteric motor neurons and from an atypical source including a potential non-neuronal origin.

194 Responsive Neurostimulation Targeting the Centromedian Nucleus of the Thalamus Improves Seizure Control in Idiopathic Generalized Epilepsy and Select Multifocal Disease

INTRODUCTION: Although more than 30% of epilepsy patients are drug resistant, many have not traditionally been considered surgical candidates due to generalized or multifocal disease. However, responsive neurostimulation (RNS) of the centromedian nucleus (CM) of the thalamus appears promising for this patient population. We present outcomes following CM RNS for patients with idiopathic generalized epilepsy (IGE) or multifocal disease, with subgroup analyses of patients’ seizure types and targeting methodology. METHODS: 21 patients with drug resistant epilepsy underwent bilateral CM RNS implantation. Electrodes were localized and volumes of tissue activation (VTAs) were modeled in Lead-DBS. Outcome measures were extracted from electronic medical records. Clinical outcomes over time were compared between patients’ seizure types and RNS recorded long-episodes, and VTA overlap with thalamic nuclei was compared between patients with direct and indirect targeting methodologies. RESULTS: For patients with at least 1-year of follow up, average seizure reduction from pre-operative baseline was 81.3%, with 3 Engel class 1 patients, 5 Engel class 2, 8 Engel class 3, and 0 Engel class 4. Average seizure reduction was greatest for non-lesional multifocal patients. Average time to worthwhile seizure reduction was 217 days, and was shortest for myoclonic IGE patients. Long-episodes were associated with seizure reduction in IGE patients only. Direct targeting led to more anterior and medial electrodes with greater VTA overlap with CM. CONCLUSIONS: CM RNS may provide significant relief for IGE and multifocal epilepsy patients. Subsets of these patient populations may particularly benefit from CM RNS. Direct targeting may facilitate more selective stimulation of the CM. The refinement of stimulation targeting, tuning of RNS system parameters, and deliberate patient selection are ongoing areas of investigation.

Enhancing Attentional Performance in Parkinson's Disease: The Impact of Combined Deep Brain Stimulation of the Substantia Nigra Pars Reticulata and the Subthalamic Nucleus

ObjectiveThe concomitant stimulation of the subthalamic nucleus and the substantia nigra pars reticulata is a promising approach to improve treatment of refractory axial symptoms in Parkinson's disease. While dual stimulation of the subthalamic nucleus and the substantia nigra pars reticulata has previously shown beneficial effects on gait, the role of the substantia nigra, a crucial component of the basal ganglia circuitry, in cognitive functions such as attention and executive control remains underexplored. This study aimed to investigate the impact of selective substantia nigra pars reticulata stimulation on attentional performance in patients receiving standard deep brain stimulation of the subthalamic nucleus. MethodsTwelve patients with bilateral subthalamic nucleus stimulation underwent computerized assessment of attention using a simple reaction time task. Reaction times were assessed under standard stimulation of the subthalamic nucleus versus simultaneous stimulation of the subthalamic nucleus and the substantia nigra pars reticulata. ResultsThe results revealed a significant improvement in reaction times during the simple reaction time task when patients received dual stimulation compared to standard stimulation. ConclusionsOur findings provide further evidence for the pivotal role of the substantia nigra pars reticulata in cognitive functions such as attention. Despite the limitations of the study, including a small sample size, our results suggest potential benefits of simultaneous deep brain stimulation of the subthalamic nucleus and the substantia nigra pars reticulata on attentional performance in patients with Parkinson's disease. Further research with larger cohorts is warranted to confirm these findings and better understand the underlying mechanisms.

Optogenetic stimulation of neurons in the anterior cingulate cortex induces changes in intravesical bladder pressure and the micturition reflex

Lower urinary tract (LUT) function is controlled by the central nervous system, including higher-order cognitive brain regions. The anterior cingulate cortex (ACC) is one of these regions, but the role of its activity in LUT function remains poorly understood. In the present study, we conducted optogenetic experiments to manipulate neural activity in mouse ACC while monitoring bladder pressure to elucidate how the activity of ACC regulates LUT function. Selective optogenetic stimulation of excitatory neurons in ACC induced a sharp increase in bladder pressure, whereas activation of inhibitory neurons in ACC prolonged the interval between bladder contractions. Pharmacological manipulation of ACC also altered bladder contractions, consistent with those observed in optogenetic experiments. Optogenetic mapping of the cortical area responsible for eliciting the increase in bladder pressure revealed that stimulation to ACC showed more potent effects than the neighboring motor cortical areas. These results suggest that ACC plays a crucial role in initiating the bladder pressure change and the micturition reflex. Thus, the balance between excitation and inhibition in ACC may regulate the reflex bidirectionally.

Development of a focused-ultrasound spectroscopic-imaging system combined with optical imaging for applying mechanical stimulation on living cells and the possibility of selective stimulation for the nucleus

Various techniques have been used for the mechanical stimulation of cells, but most of them require direct contact with cells and can damage them. Here, we propose a focused ultrasound technique for non-invasive localized mechanical stimulation in a culture environment. First, we develop the focused ultrasound system for establishing the spectroscopic images and for applying mechanical stimulation combined with optical observation. Second, we construct the acoustic-absorption spectroscopic images of human mesenchymal stem cells, leading to the finding of the ultrasound absorption band of the nucleus around 130 MHz caused by its resonance. Finally, we find that the nucleus less absorbs ultrasound before the cell division, supporting our view that the acoustic absorption band is related to the resonance of the nucleus. Our results show the potential for realizing selective and effective mechanical stimulation for nuclei non-invasively, which is critically important for a deeper understanding of mechanobiology.