-
The Journal of Neuroscience : the... Nov 2014In myelinated peripheral axons, Kv1 potassium channels are clustered at the juxtaparanodal region and at an internodal line located along the mesaxon and below the...
In myelinated peripheral axons, Kv1 potassium channels are clustered at the juxtaparanodal region and at an internodal line located along the mesaxon and below the Schmidt-Lanterman incisures. This polarized distribution is controlled by Schwann cells and requires specific cell adhesion molecules (CAMs). The accumulation of Kv1 channels at the juxtaparanodal region depends on the presence of Caspr2 at this site, as well as on the presence of Caspr at the adjacent paranodal junction. However, the localization of these channels along the mesaxonal internodal line still persists in the absence of each one of these CAMs. By generating mice lacking both Caspr and Caspr2 (caspr(-/-)/caspr2(-/-)), we now reveal compensatory functions of the two proteins in the organization of the axolemma. Although Kv1 channels are clustered along the inner mesaxon and in a circumferential ring below the incisures in the single mutants, in sciatic nerves of caspr(-/-)/caspr2(-/-) mice, these channels formed large aggregates that were dispersed along the axolemma, demonstrating that internodal localization of Kv1 channels requires either Caspr or Caspr2. Furthermore, deletion of both Caspr and Caspr2 also resulted in widening of the nodes of Ranvier, suggesting that Caspr2 (which is present at paranodes in the absence of Caspr) can partially compensate for the barrier function of Caspr at this site even without the formation of a distinct paranodal junction. Our results indicate that Caspr and Caspr2 are required for the organization of the axolemma both radially, manifested as the mesaxonal line, and longitudinally, demarcated by the nodal domains.
Topics: Animals; Axons; Cell Adhesion Molecules, Neuronal; Kv1.2 Potassium Channel; Membrane Proteins; Mice; Nerve Tissue Proteins; Protein Transport; Ranvier's Nodes
PubMed: 25378149
DOI: 10.1523/JNEUROSCI.3369-14.2014 -
Journal of the Neurological Sciences Feb 2018Workers exposed to aerosolized brain in a swine-processing plant developed immune-mediated polyradiculoneuropathy (IP) possibly triggered by an immune response.
IMPORTANCE
Workers exposed to aerosolized brain in a swine-processing plant developed immune-mediated polyradiculoneuropathy (IP) possibly triggered by an immune response.
OBJECTIVE
Immunohistochemistry results were correlated with electrophysiological variables to examine the immunopathogenesis of this disorder.
DESIGN/SETTING
Laboratory studies used normal nerve tissue that was exposed to sera from 12 IP patients; 10 exposed controls; and 10 unexposed controls. Clinical and electrophysiological data from IP patients were obtained from medical record reviews.
MAIN OUTCOME MEASURES
Analysis included electromyography results of IP patients and nerve conduction studies examining CMAP amplitude, distal motor latency, motor conduction velocity, F-wave latency, sensory nerve action potential amplitude, and sensory nerve conduction velocity. Case and control results were compared relative to distance from exposure.
RESULTS
Electrodiagnostic findings revealed prolongation of the distal and f-wave latencies suggestive of demyelination at the level of the nerve root and distal nerve terminals. Immunohistochemical results identified an antibody to the peripheral nerve, with staining at the level of the axolemma. Thus, IP may be a primary axonopathy with secondary paranodal demyelination causing the conduction changes. Staining of the distal and proximal portions of the nerve appears consistent with easier access through the blood-nerve barrier.
CONCLUSIONS AND RELEVANCE
IP is an immune-mediated neuropathy related to antibodies to an axon-based antigen on peripheral nerves. Secondary paranodal demyelination is likely. Further studies to identify the primary axonal antigenic target would be useful.
Topics: Abattoirs; Adult; Evoked Potentials, Motor; Female; Humans; Immunohistochemistry; Male; Middle Aged; Muscle, Skeletal; Myelin Sheath; Nerve Tissue Proteins; Neural Conduction; Polyradiculoneuropathy; Reaction Time; Retrospective Studies; Young Adult
PubMed: 29406910
DOI: 10.1016/j.jns.2017.12.005 -
Brain : a Journal of Neurology Apr 2017See Saporta and Shy (doi:10.1093/awx048) for a scientific commentary on this article.Effective bidirectional signalling between axons and Schwann cells is essential for...
See Saporta and Shy (doi:10.1093/awx048) for a scientific commentary on this article.Effective bidirectional signalling between axons and Schwann cells is essential for both the development and maintenance of peripheral nerve function. We have established conditions by which human induced pluripotent stem cell-derived sensory neurons can be cultured with rat Schwann cells, and have produced for the first time long-term and stable myelinating co-cultures with human neurons. These cultures contain the specialized domains formed by axonal interaction with myelinating Schwann cells, such as clustered voltage-gated sodium channels at the node of Ranvier and Shaker-type potassium channel (Kv1.2) at the juxtaparanode. Expression of type III neuregulin-1 (TIIINRG1) in induced pluripotent stem cell-derived sensory neurons strongly enhances myelination, while conversely pharmacological blockade of the NRG1-ErbB pathway prevents myelination, providing direct evidence for the ability of this pathway to promote the myelination of human sensory axons. The β-secretase, BACE1 is a protease needed to generate active NRG1 from the full-length form. Due to the fact that it also cleaves amyloid precursor protein, BACE1 is a therapeutic target in Alzheimer's disease, however, consistent with its role in NRG1 processing we find that BACE1 inhibition significantly impairs myelination in our co-culture system. In order to exploit co-cultures to address other clinically relevant problems, they were exposed to anti-disialosyl ganglioside antibodies, including those derived from a patient with a sensory predominant, inflammatory neuropathy with mixed axonal and demyelinating electrophysiology. The co-cultures reveal that both mouse and human disialosyl antibodies target the nodal axolemma, induce acute axonal degeneration in the presence of complement, and impair myelination. The human, neuropathy-associated IgM antibody is also shown to induce complement-independent demyelination. Myelinating co-cultures using human induced pluripotent stem cell-derived sensory neurons thus provide insights into the cellular and molecular specialization of axoglial signalling, how pharmacological agents may promote or impede such signalling and the pathogenic effects of ganglioside antibodies.awx012media15372351982001.
Topics: Adult; Animals; Antibodies, Anti-Idiotypic; Cell Differentiation; Coculture Techniques; ErbB Receptors; Female; Humans; Immunoglobulin G; Mice; Myelin Sheath; Neural Stem Cells; Neuregulin-1; Peripheral Nervous System; Rats; Schwann Cells; Sensory Receptor Cells; Transduction, Genetic
PubMed: 28334857
DOI: 10.1093/brain/awx012 -
Glia Sep 2020Finding causative genetic mutations is important in the diagnosis and treatment of hereditary peripheral neuropathies. This study was conducted to find new genes...
Finding causative genetic mutations is important in the diagnosis and treatment of hereditary peripheral neuropathies. This study was conducted to find new genes involved in the pathophysiology of hereditary peripheral neuropathy. We identified a new mutation in the EBP50 gene, which is co-segregated with neuropathic phenotypes, including motor and sensory deficit in a family with Charcot-Marie-Tooth disease. EBP50 is known to be important for the formation of microvilli in epithelial cells, and the discovery of this gene mutation allowed us to study the function of EBP50 in the nervous system. EBP50 was strongly expressed in the nodal and paranodal regions of sciatic nerve fibers, where Schwann cell microvilli contact the axolemma, and at the growth tips of primary Schwann cells. In addition, EBP50 expression was decreased in mouse models of peripheral neuropathy. Knockout mice were used to study EBP50 function in the peripheral nervous system. Interestingly motor function deficit and abnormal histology of nerve fibers were observed in EBP50 heterozygous mice at 12 months of age, but not 3 months. in vitro studies using Schwann cells showed that NRG1-induced AKT activation and migration were significantly reduced in cells overexpressing the I325V mutant of EBP50 or cells with knocked-down EBP50 expression. In conclusion, we show for the first time that loss of function due to EBP50 gene deficiency or mutation can cause peripheral neuropathy.
Topics: Animals; Charcot-Marie-Tooth Disease; Mice; Mice, Knockout; Mutation; Peripheral Nerves; Peripheral Nervous System
PubMed: 32077526
DOI: 10.1002/glia.23805 -
Frontiers in Molecular Biosciences 2021Around half of the traumatic brain injuries are thought to be axonal damage. Disruption of the cellular membranes, or alternatively cytoskeletal damage has been...
Around half of the traumatic brain injuries are thought to be axonal damage. Disruption of the cellular membranes, or alternatively cytoskeletal damage has been suggested as possible injury trigger. Here, we have used molecular models to have a better insight on the structural and mechanical properties of axon sub-cellular components. We modelled myelin sheath and node of Ranvier as lipid bilayers at a coarse grained level. We built ex-novo a model for the myelin. Lipid composition and lipid saturation were based on the available experimental data. The model contains 17 different types of lipids, distributed asymmetrically between two leaflets. Molecular dynamics simulations were performed to characterize the myelin and node-of-Ranvier bilayers at equilibrium and under deformation and compared to previous axolemma simulations. We found that the myelin bilayer has a slightly higher area compressibility modulus and higher rupture strain than node of Ranvier. Compared to the axolemma in unmyelinated axon, mechanoporation occurs at 50% higher strain in the myelin and at 23% lower strain in the node of Ranvier in myelinated axon. Combining the results with finite element simulations of the axon, we hypothesizes that myelin does not rupture at the thresholds proposed in the literature for axonal injury while rupture may occur at the node of Ranvier. The findings contribute to increases our knowledge of axonal sub-cellular components and help to understand better the mechanism behind axonal brain injury.
PubMed: 34250015
DOI: 10.3389/fmolb.2021.669897 -
Journal of Neurotrauma May 2016Traumatic brain injury (TBI) is a health threat that affects every year millions of people involved in motor vehicle and sporting accidents, and thousands of soldiers in...
Traumatic brain injury (TBI) is a health threat that affects every year millions of people involved in motor vehicle and sporting accidents, and thousands of soldiers in battlefields. Diffuse axonal injury (DAI) is one of the most frequent types of TBI leading to death. In DAI, the initial traumatic event is followed by a cascade of biochemical changes that take time to develop in full, so that symptoms may not become apparent until days or weeks after the original injury. Hence, DAI is a dynamic process, and the opportunity exists to prevent its progression provided the initial trauma can be predicted at the molecular level. Here, we present preliminary evidence from micro-finite element (FE) simulations that the mechanical response of central nervous system myelinated fibers is dependent on the axonal diameter, the ratio between axon diameter and fiber diameter (g-ratio), the microtubules density, and the cholesterol concentration in the axolemma and myelin. A key outcome of the simulations is that there is a significant difference between the overall level of strain in a given axonal segment and the level of local strain in the Ranvier nodes contained in that segment, with the nodal strain being much larger than the total strain. We suggest that the acquisition of this geometric and biochemical information by means of already available high resolution magnetic resonance imaging techniques, and its incorporation in current FE models of the brain will enhance the models capacity to predict the site and magnitude of primary axonal damage upon TBI.
Topics: Axons; Brain Injuries, Traumatic; Cholesterol; Diffuse Axonal Injury; Finite Element Analysis; Humans; Models, Biological; Ranvier's Nodes; Stress, Mechanical
PubMed: 26393780
DOI: 10.1089/neu.2015.4007 -
Investigative Ophthalmology & Visual... Feb 2015Decreased ATP correlates with intraocular pressure exposure in the optic nerves of mice with glaucoma. To understand what underlies this energy deficit, we examined...
PURPOSE
Decreased ATP correlates with intraocular pressure exposure in the optic nerves of mice with glaucoma. To understand what underlies this energy deficit, we examined mitochondria in the myelinated optic nerve axons of the DBA/2J mouse, a model of glaucoma secondary to iris pigment disease, and the DBA/2(wt-gpnmb) control strain.
METHODS
Mitochondrial length, width, surface area, and health status were measured in 30 electron microscopic fields within the myelinated portion of optic nerves from DBA/2J and DBA/2(wt-gpnmb) mice at 3, 6, and 10 months of age. Protein was isolated from optic nerve for analysis of PINK1, Parkin, LC3-I and -II, and lysosome-associated membrane protein 1 (LAMP1) by Western blot.
RESULTS
The number of mitochondria in DBA/2J optic nerve was increased, and they had significantly smaller surface area. Mitochondria in DBA/2J were closer to the axolemma, more spatially isolated, and their cristae were more disrupted at every age group as compared to DBA/2(wt-gpnmb). Autophagosomes were significantly increased in DBA/2J optic nerve at all ages. Protein analysis showed higher LC3-II to LC3-I ratio in aged DBA/2J optic nerve than in DBA/2(wt-gpnmb). PINK1 and Parkin levels were not statistically different across age groups. LAMP1 was significantly decreased in the aged DBA/2J optic nerve.
CONCLUSIONS
Decreased surface area, combined with reduced oxidative capacity in mitochondria from the aged DBA/2J axon, indicate that mitochondrial pathology may contribute to the energy deficit in glaucomatous optic nerve. Though autophagosomes were increased in DBA/2J optic nerve, the increased mitochondria and decreased LAMP1 suggest deteriorating mitochondria are not being efficiently recycled by mitophagy.
Topics: Adenosine Triphosphate; Age Factors; Animals; Axons; Disease Models, Animal; Mice; Mice, Inbred DBA; Microscopy, Electron; Mitochondria; Mitophagy; Nerve Fibers, Myelinated; Optic Nerve
PubMed: 25655803
DOI: 10.1167/iovs.14-16126 -
Journal of Biomechanical Engineering Dec 2019Diffuse axonal injury (DAI) is the pathological consequence of traumatic brain injury (TBI) that most of all requires a multiscale approach in order to be, first,...
Diffuse axonal injury (DAI) is the pathological consequence of traumatic brain injury (TBI) that most of all requires a multiscale approach in order to be, first, understood and then possibly prevented. While in fact the mechanical insult usually happens at the head (or macro) level, the consequences affect structures at the cellular (or microlevel). The quest for axonal injury tolerances has so far been addressed both with experimental and computational approaches. On one hand, the experimental approach presents challenges connected to both temporal and spatial resolution in the identification of a clear axonal injury trigger after the application of a mechanical load. On the other hand, computational approaches usually consider axons as homogeneous entities and therefore are unable to make inferences about their viability, which is thought to depend on subcellular damages. Here, we propose a computational multiscale approach to investigate the onset of axonal injury in two typical experimental scenarios. We simulated single-cell and tissue stretch injury using a composite finite element axonal model in isolation and embedded in a matrix, respectively. Inferences on axonal damage are based on the comparison between axolemma strains and previously established mechanoporation thresholds. Our results show that, axons embedded in a tissue could withstand higher deformations than isolated axons before mechanoporation occurred and this is exacerbated by the increase in strain rate from 1/s to 10/s.
PubMed: 31556941
DOI: 10.1115/1.4044953 -
Frontiers in Neurology 2018Diffuse axonal injury (DAI) occurs as a result of the transmission of rapid dynamic loads from the head to the brain and specifically to its neurons. Despite being one...
Diffuse axonal injury (DAI) occurs as a result of the transmission of rapid dynamic loads from the head to the brain and specifically to its neurons. Despite being one of the most common and most deleterious types of traumatic brain injury (TBI), the inherent cell injury mechanism is yet to be understood. Experimental observations have led to the formulation of different hypotheses, such as mechanoporation of the axolemma and microtubules (MTs) breakage. With the goal of singling out the mechanical aspect of DAI and to resolve the ambiguity behind its injury mechanism, a composite finite element (FE) model of a representative volume of an axon was developed. Once calibrated and validated against published experimental data, the axonal model was used to simulate injury scenarios. The resulting strain distributions along its components were then studied in dependence of strain rate and of typical modeling choices such as the applied MT constraints and tau proteins failure. Results show that oversimplifying the MT bundle kinematic entails a systematic attenuation ( = 2.33) of the computed maximum MT strain. Nevertheless, altogether, results support the hypothesis of axolemma mechanoporation as a cell-injury trigger. Moreover, for the first time the interconnection between the axolemma and the MT bundle is shown to play a role in damage localization. The proposed FE axonal model is a valuable tool to understand the axonal injury mechanism from a mechanical perspective and could be used in turn for the definition of well-informed injury criteria at the tissue and organ level.
PubMed: 30127763
DOI: 10.3389/fneur.2018.00643 -
Frontiers in Cellular Neuroscience 2018It is today widely accepted that several types of high molecular weight (MW) neurotransmitters produced by neurons are synthesized at the cell body, selectively stored...
It is today widely accepted that several types of high molecular weight (MW) neurotransmitters produced by neurons are synthesized at the cell body, selectively stored within large dense core vesicles (LDCVs) and anterogradely transported to terminals where they elicit their biological role(s). Among these molecules there are neuropeptides and neurotrophic factors, the main focus of this perspective article. I here first provide a brief resume of the state of art on neuronal secretion, with primary emphasis on the molecular composition and mechanism(s) of filling and release of LDCVs. Then, I discuss the perspectives and future directions of research in the field as regarding the synthesis and storage of multiple high MW transmitters in LDCVs and the possibility that a selective sorting of LDCVs occurs along different neuronal processes and/or their branches. I also consider the ongoing discussion that diverse types of neurons may contain LDCVs with different sets of integral proteins or dial in a different fashion with LDCVs containing the same cargo. In addition, I provide original data on the size of LDCVs in rat dorsal root ganglion neurons and their central terminals in the spinal cord after immunogold labeling for calcitonin gene-related peptide (CGRP), neuropeptide K, substance P, neurokinin A or somatostatin. These data corroborate the idea that, similarly to endocrine cells, LDCVs undergo a process of maturation which involves a homotypic fusion followed by a reduction in size and condensation of cargo. They also give support to the conjecture that release at terminals occurs by cavicapture, a process of partial fusion of the vesicle with the axolemma, accompanied by depletion of cargo and diminution of size.
PubMed: 30186121
DOI: 10.3389/fncel.2018.00272