Correlation of cell body size, axon size, and signal conduction velocity for individually labelled dorsal root ganglion cells in the cat

Measurements of cell body and peripheral and central axon sizes were made for primary sensory neurons outlined by the intracellular injection of HRP. Conduction velocities were also measured on the outlined processes. The sensory neurons were then subdivided into A and C cells on the basis of the conduction velocity of the impulses carried by the processes of these cells. Central processes of both A and C cells are smaller than the peripheral processes, but the size differential is greater for the C cells. For A cells there is a linear relation between the size of the peripheral axon and the conduction velocity of the impulses carried by these axons, but the confidence limits are wide. For C cells there is a linear relation between the size of the central process and conduction velocity of the impulses carried by the processes, but for the peripheral processes two aberrant processes resulted in no correlation between process size and conduction velocity. For A cells, the size of the central and peripheral processes and the conduction velocity of the impulses carried by the peripheral processes are linearly correlated with cell body size. By contrast no such correlations can be demonstrated for C cells. This presumably implies an important difference in that the size of the cell body is correlated with axon size and impulse conduction velocity for A cells but not for C cells. A widely accepted generalization is that large sensory cells give rise to myelinated axons and small sensory cells to unmyelinated axons. In this study, myelinated and unmyelinated are defined on the basis of impulse conduction velocity. For those cells that are clearly large (greater than 50 microns in diameter), the conduction velocity of the impulses carried by their processes is always greater than 2.5 m/s, and for those cells that are clearly small (less than 35 microns in diameter), the conduction velocity is always less than 2.5 m/s. Thus for these cells the above generalization holds. For the intermediate-sized cells (35-50 microns), however, the size of the cell body bears no predictable relation to the conduction velocity of the impulses carried by those processes, and thus to whether the axons are myelinated or unmyelinated. Thus the above generalization does not hold for this intermediate group of cells, and since there are many cells in this size range, we feel that the generalization that large cells give rise to myelinated axons and small cells to unmyelinated axons is an oversimplification.     

Acetylcholinesterase antibody treatment results in neurite detachment and reduced outgrowth from cultured neurons: Further evidence for a cell adhesive role for neuronal acetylcholinesterase

Data from our laboratory and others demonstrate that acetylcholinesterase (AChE) is expressed transiently by neurons during periods of neurite outgrowth preceding synaptogenesis, suggesting an extrasynaptic function for this molecule. These findings, along with reports that AChE shares amino acid sequence homology and structural similarities with known cell adhesion molecules, have led to the theory that, during development, AChE may exert a morphogenic effect through cell adhesion. To further test this hypothesis, we have examined the effects of an AChE monoclonal antibody (MAB304) on neurite outgrowth in primary cultures of rat dorsal root ganglion (DRG) neurons. Short-term, high-concentration antibody treatment produced a rapid detachment of established DRG neurites, which was followed by regrowth upon removal of the antibody from the culture medium. This effect appeared to be site-specific, because other AChE antibodies that were able to detect AChE immunocytochemically failed to produce this disadhesion. Long-term, low-concentration antibody exposure produced a 50% reduction in total area of outgrowth, in which neurites were more densely packed and interlaced compared with the neurites in control cultures. These results extend our previous observations on the outgrowth perturbing effects of AChE inhibitor treatment and provide further evidence that AChE may support neurite outgrowth through a cell adhesive role. J. Neurosci. Res. 53:454–464, 1998. © 1998 Wiley-Liss, Inc.     

Percutaneous Pulsed Radiofrequency Treatment of the Cervical Dorsal Root Ganglion in the Treatment of Chronic Cervical Pain Syndromes: A Clinical Audit

Cervicogenic headache and cervicobrachialgia are frequent diagnoses of chronic cervical pain. After failure of conservative treatment, an interventional approach may be indicated in the absence of any indication for causal surgical treatment. The pulsed radiofrequency (PRF) technique exposes the nerve to a high‐frequency electric field while the temperature of the electrode tip does not exceed 42°C. This method is thought to be nondestructive and almost free of neurologic side effects and complications. Our extended pilot study was performed to confirm the perceived efficacy of PRF for short‐ and long‐term relief of chronic cervical pain. We carried out a clinical audit of the first 18 patients treated with PRF at the cervical dorsal root ganglion. An independent evaluator reviewed the medical records. Patients with good clinical results at 8 weeks were evaluated for long‐term effect (> 6 months), based on a 7‐point Likert scale. Thirteen patients (72%) showed short‐term clinical success (≥ 50% pain relief). Mean follow‐up was 19.4 months (SD 8.9 months), maximum 2.5 years. The duration of satisfactory pain relief (6 or 7 on the Likert scale) varied between 2 and over 30 months, with a mean duration of 9.2 months (SD 11.2 months). Kaplan‐Meier analysis illustrated that 50% of patients experienced success 3 months after treatment. We could not identify predictive variables for clinical outcome. None of the patients reported post‐treatment neuritis or other adverse events. To our knowledge, this is the first documented series of chronic cervical pain syndromes treated with PRF. Satisfactory pain relief of at least 50% was achieved in 13 of 18 (72%) patients at 8 weeks. More than one year after treatment, six patients (33%) continue to rate treatment outcome as good or very good. No side effects were reported. j     

Glial-mediated neuroprotection: evidence for the protective role of the NO-cGMP pathway via neuron-glial communication in the peripheral nervous system

The NO-cGMP pathway has emerged as a neuroprotective signaling system involved in communication between neurons and glia. We have previously shown that axotomy or nerve growth factor (NGF)-deprivation of dorsal root ganglion (DRG) neurons leads to increased production of NO and at the same time an increase in cGMP production in their satellite glia cells. Blockade of NO or its receptor, the cGMP synthesizing enzyme soluble guanylate cyclase (sGC), results in apoptosis of neurons and glia. We now show that co-culture of neonatal DRG neurons with either Schwann cells pre-treated with an NO donor or a membrane-permeant cGMP analogue; or neurons maintained in the medium from Schwann cell cultures treated in the same way, prevents neuronal apoptosis. Both NO donor and cGMP treatment of Schwann cells results in synthesis of NGF and NT3. Furthermore, if the Schwann cells are previously infected with adenoviral vectors expressing a dominant negative sGC mutant transgene, treatment of these Schwann cells with an NO donor now fails to prevent neuronal apoptosis. Schwann cells treated in this way also fail to express neither cGMP nor neurotrophins. These findings suggest NO-sGC-cGMP-mediated NGF and NT3 synthesis by Schwann cells protect neurons.     

Targeting cell surface receptors for axon regeneration in the central nervous system

Axon regeneration in the CNS is largely unsuccessful due to excess inhibitory extrinsic factors within lesion sites together with an intrinsic inability of neurons to regrow following injury. Recent work demonstrates that forced expression of certain neuronal transmembrane receptors can recapitulate neuronal growth resulting in successful growth within and through inhibitory lesion environments. More specifically, neuronal expression of integrin receptors such as alpha9beta1 integrin which binds the extracellular matrix glycoprotein tenascin-C, trk receptors such as trk B which binds the neurotrophic factor BDNF, and receptor PTPσ which binds chondroitin sulphate proteoglycans, have all been show to significantly enhance regeneration of injured axons. We discuss how reintroduction of these receptors in damaged neurons facilitates signalling from the internal environment of the cell with the external environment of the lesion milieu, effectively resulting in growth and repair following injury. In summary, we suggest an appropriate balance of intrinsic and extrinsic factors are required to obtain substantial axon regeneration.     

Retinoic acid responsive gene product,midkine, has neurotrophic functions for mouse spinal cord and dorsal root ganglion neurons in culture

Midkine (MK) is the product of a retinoic acid responsive gene and is a member of a new family of heparin-binding growth factors. Neurotrophic effects of MK were examined using cultured spinal cord and dorsal root ganglion (DRG) neurons derived from fetal mouse. MK, which was added to the culture medium at concentrations of 1–100 ng/ml, promoted survival of both types of neurons approximately 5-fold after 7 days in culture. For spinal cord neurons, the increased survival was reflected in an increase of choline acetyltransferase activity. MK also promoted neurite extension in spinal cord (2-fold) and DRG (1.7-fold) neurons. The survival-promoting activity of MK to these neurons was comparable to that of basic fibroblast growth factor (bFGF) and leukemia inhibitory factor (LIF). In spite of its significant effects on fetal neurons, MK was ineffective in sustaining survival of DRG neurons derived from postnatal mice. From these results, we conclude that MK is a neurotrophic factor to embryonic spinal cord and DRG neurons, and we propose that MK plays a significant role in embryogenesis of the nervous system. © 1993 Wiley-Liss, Inc.     

Chondroitin Sulphate Proteoglycans in the Rat Brain: Candidates for Axon Barriers of Sensory Neurons and the Possible Modification by Laminin of their Actions

The addition of chondroitin sulphate proteoglycans (CSPGs), purified from the rat brain, to the culture medium of PC12D cells inhibited their proliferation and neurite outgrowth. Therefore, we investigated the effects of several extracellular components on the inhibitory actions of CSPGs on PC12D cells, as well as their immunocytochemical distribution in the rat embryo to determine whether the findings in vitro could be reproduced in vivo. Coating of the substratum with polylysine was necessary for the appearance of the inhibitory effects of brain CSPGs on PC12D cells. The additional pretreatment of polylysine-coated dishes with laminin or fibronectin promoted the outgrowth of neurites from PC12D cells. Laminin and fibronectin, but not collagen (types I and IV) and CELL-TAK (cell adhesion molecules), prevented the inhibitory effects of brain CSPGs in a concentration-dependent manner. Doses producing 50% reduction by laminin (or fibronectin) of the CSPG effects were 1.5 (or 25) μg/ml for neurite outgrowth and 2.2 (or 28) μg/ml for proliferation. The ratio of dish-attached CSPGs to laminin necessary for 50% reduction was about ∼50:l (wt/wt). Laminin from any source had the same effect. Brain CSPGs also obviously impeded the growth of fibres from dorsal root ganglion explants and primary cultured dorsal root ganglion neurons. Neurocan (a major CSPG in the brain)-like immunoreactivity was detected in the boundary caps and roof plate in the rat embryo at 13.5 days of gestation, when DRG neurons were extending their axons to the neural tube. The distributions of laminin and tenascin appeared, respectively, to be slightly and considerably different from that of neurocan. These results suggest that brain CSPGs can generate barriers to the growth of axons from the dorsal root ganglion and that the ratio of CSPGs to laminin may be important in regulating such growth.     

Peripheral sensory nerve fibers that dichotomize to supply the brachium and the pericardium in the rat: a possible morphological explanation for referred cardiac pain?

Two fluorescent dyes, ‘True Blue’ and ‘Diamidino Yellow’ were injected, respectively, into the pericardial sac and into the medial brachial cutaneous nerve or subcutaneously into the medial side of the brachium. Double-labelling was observed in ipsilateral dorsal root ganglia neurons of spinal cord segments C8, T1 and T2, indicating that dichotomizing afferent fibers supply both the pericardium and the brachium. This finding provides a possible morphological explanation for referred cardiac pain.     

TRPV1, but not P2X, requires cholesterol for its function and membrane expression in rat nociceptors

We examined the importance of membrane cholesterol for the function and expression of TRPV1 (vanilloid receptor subtype 1) and P2X(3) in adult rat dorsal root ganglion (DRG) neurons. Cholesterol, an essential component of lipid rafts, was depleted using methyl beta-cyclodextrin (MCD). We found that MCD significantly reduced TRPV1-mediated capsaicin- and proton-activated currents. By contrast, inward currents activated by alpha,beta-methylene ATP (alpha,beta-meATP), a non-hydrolysable ATP analogue, were not altered. Immunoreactivity for TRPV1, but not P2X(3), in the plasma membrane was markedly reduced by MCD. A reduction of TRPV1 protein in membrane fractions was found following cholesterol depletion. Our data show that the level of cholesterol determines the activity and the amount of membrane TRPV1, suggesting that TRPV1 might be localized in cholesterol-rich microdomains in nociceptors. The differential dependence on the membrane cholesterol of TRPV1 and P2X(3) may have physiological significance in nociception during inflammation.     

Common data elements in epilepsy research: development and implementation of the NINDS epilepsy CDE project

The Common Data Element (CDE) Project was initiated in 2006 by the National Institute of Neurological Disorders and Stroke (NINDS) to develop standards for performing funded neuroscience-related clinical research. CDEs are intended to standardize aspects of data collection; decrease study start-up time; and provide more complete, comprehensive, and equivalent data across studies within a particular disease area. Therefore, CDEs will simplify data sharing and data aggregation across NINDS-funded clinical research, and where appropriate, facilitate the development of evidenced-based guidelines and recommendations. Epilepsy-specific CDEs were established in nine content areas: (1) Antiepileptic Drugs (AEDs) and Other Antiepileptic Therapies (AETs), (2) Comorbidities, (3) Electrophysiology, (4) Imaging, (5) Neurological Exam, (6) Neuropsychology, (7) Quality of Life, (8) Seizures and Syndromes, and (9) Surgery and Pathology. CDEs were developed as a dynamic resource that will accommodate recommendations based on investigator use, new technologies, and research findings documenting emerging critical disease characteristics. The epilepsy-specific CDE initiative can be viewed as part of the larger international movement toward "harmonization" of clinical disease characterization and outcome assessment designed to promote communication and research efforts in epilepsy. It will also provide valuable guidance for CDE improvement during further development, refinement, and implementation. This article describes the NINDS CDE Initiative, the process used in developing Epilepsy CDEs, and the benefits of CDEs for the clinical investigator and NINDS.     

A Comprehensive Practice Guideline for Magnetic Resonance Imaging Compatibility in Implanted Neuromodulation Devices

ObjectivesThe evolution of neuromodulation devices in order to enter magnetic resonance imaging (MRI) scanners has been one of understanding limitations, engineering modifications, and the development of a consensus within the community in which the FDA could safely administer labeling for the devices. In the initial decades of neuromodulation, it has been contraindicated for MRI use with implanted devices. In this review, we take a comprehensive approach to address all the major products currently on the market in order to provide physicians with the ability to determine when an MRI can be performed for each type of device implant.Materials and MethodsWe have prepared a narrative review of MRI guidelines for currently marketed implanted neuromodulation devices including spinal cord stimulators, intrathecal drug delivery systems, peripheral nerve stimulators, deep brain stimulators, vagal nerve stimulators, and sacral nerve stimulators. Data sources included relevant literature identified through searches of PubMed, MEDLINE/OVID, SCOPUS, and manual searches of the bibliographies of known primary and review articles, as well as manufacturer-provided information.ResultsGuidelines and recommendations for each device and their respective guidelines for use in and around MR environments are presented.ConclusionsThis is the first comprehensive guideline with regards to various devices in the market and MRI compatibility from the American Society of Pain and Neuroscience.     

Three-dimensional organisation of mitochondrial clusters in regenerating dorsal root ganglion (DRG) neurons from neonatal rats: evidence for mobile mitochondrial pools

Abstract   We report for the first time the rearrangement of mitochondrial arrays in developing dorsal root ganglion (DRG) neurons isolated from neonatal rats in culture. Neurons were loaded with the mitochondria-specific fluorescent dye JC-1, and three-dimensional (3D) reconstruction of mitochondrial fluorescence was performed by confocal laser sectioning in fresh neurons and neurons kept in culture up to a week. We found that after 24 hours the mitochondria become reorganised to form clusters in the axonal hillocks. Axonal extension and neuronal network formation coincided with a redistribution of the mitochondrial clusters. In the extended axons the mitochondria become spaced along the axonal length; however, they formed clusters in the branch points and growth cones. We conclude that the initial clusters of mitochondria may be storage pools of mobile mitochondria able to be mobilised to provide energy for axonal transport during neuronal regeneration and neuronal outgrowth. These findings may have relevance to the rate of axonal regeneration and axonal transport in adult DRG neurons, and neuronal polarisation and axonal outgrowth regulation in developing DRG neurons.     

Measuring the rate of progression in Friedreich ataxia: Implications for clinical trial design

Friedreich ataxia is an autosomal recessive neurodegenerative disorder characterized by ataxia of all four limbs, dysarthria, and arreflexia. A variety of measures are currently used to quantify disease progression, including the Friedreich Ataxia Rating Scale, examiner‐rated functional disability scales, self‐reported activities of daily living and performance measures such as the timed 25‐foot walk, 9‐hole pegboard test, PATA speech test, and low‐contrast letter acuity vision charts. This study examines the rate of disease progression over one and two years in a cohort of 236 Friedreich ataxia patients using these scales and performance measure composites. The Friedreich Ataxia Rating Scale and performance‐measure composites captured disease progression, with a greater sensitivity to change over 2 years than over 1 year. The measures differed in their sensitivity to change and in possible bias. These results help to establish norms for progression in FRDA that can be useful in measuring the long‐term success of therapeutic agents and defining sample‐size calculations for double‐blind clinical trials. © 2010 Movement Disorder Society     

Efficient use of clinical EEG data for deep learning in epilepsy

ObjectiveAutomating detection of Interictal Epileptiform Discharges (IEDs) in electroencephalogram (EEG) recordings can reduce the time spent on visual analysis for the diagnosis of epilepsy. Deep learning has shown potential for this purpose, but the scarceness of expert annotated data creates a bottleneck in the process.MethodsWe used EEGs from 50 patients with focal epilepsy, 49 patients with generalized epilepsy (IEDs were visually labeled by experts) and 67 controls. The data was filtered, downsampled and cut into two second epochs. We increased the number of input samples containing IEDs through temporal shifting and using different montages. A VGG C convolutional neural network was trained to detect IEDs.ResultsUsing the dataset with more samples, we reduced the false positive rate from 2.11 to 0.73 detections per minute at the intersection of sensitivity and specificity. Sensitivity increased from 63% to 96% at 99% specificity. The model became less sensitive to the position of the IED in the epoch and montage.ConclusionsTemporal shifting and use of different EEG montages improves performance of deep neural networks in IED detection.SignificanceDataset augmentation can reduce the need for expert annotation, facilitating the training of neural networks, potentially leading to a fundamental shift in EEG analysis.     

A public health approach to clinical therapeutics in psychiatry: directions for new research

The mental health field is transforming the culture of treatment research by moving from a narrow regulatory model geared to drug approval and registration to a more inclusive public health model. Thus, whereas regulatory antidementia trials will exclude patients with psychiatric or neurologic symptoms or substance abuse and require them to be physically healthy and living with a caregiver, ie, 90% of the presenting Alzheimer population, the public health model promises to improve patient care by addressing the types of practical questions and functional outcomes typically the concern of clinicians: Does treatment enhance function? How can we keep people well once they have been made well? Why do treatments not work as well in practice as in clinical trials? Public health studies are conducted in the world of actual practice with time-pressured clinicians taking care of large numbers of patients with uncertain clinical presentations, complex comorbidities, and varying degrees of interference with ideal levels of compliance. The exclusive focus on symptoms is expanded to include outcomes related to issues of function, disability, morbidity, mortality, resource use, and quality of life. Highly controlled efficacy research is still needed to establish treatment merit, but efficacy now marks only the beginning of the process of inquiry.     

From research and development to practice-based evidence: clinical governance initiatives in a service for adults with mild intellectual disability and mental health needs

Background Practice‐based evidence represents the contribution of practitioners who utilize research methodologies to examine the quality of their clinical practice and service provision. Methods The present paper describes the evolution of a routine practice‐based evidence system (PBES) via four phases of research and development. The four phases are described, as is their relevance to assessment and intervention with regard to the mental health problems of people with mild intellectual disability. Phase four describes the development of a routine PBES. Results The PBES is capable of profiling the individual mental health needs of service users and examining service effectiveness and quality at an organizational level. Conclusions The PBES is discussed according to its current utility and possible directions for future development. The system is presented as an example of clinical governance that could be utilized by multidisciplinary teams to develop and maintain an organizational culture of quality.