Differential binding patterns of anti-sulfatide antibodies to glial membranes

Meehan, Gavin R.; McGonigal, Rhona; Cunningham, Madeleine E.; Wang, Yu‐Zhong; Barrie, Jennifer A.; Halstead, Susan K.; Gourlay, Dawn; Yao, Denggao; Willison, Hugh J.

doi:10.1016/j.jneuroim.2018.07.004

Cited by 8 publications

(10 citation statements)

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“…To firmly establish that the observed disruptions to paranodal proteins were not partly a confounding consequence of structural dysfunction secondary to abnormal ganglioside profiles in the axonal compartment of transgenic Glial mice, a complement-fixing antibody against the myelin glycolipid sulfatide ( 27 ) was used to target the paranodal loop glial membrane in WT mice in order to look for similar perturbations. As seen with the anti-GM1 antibody in Glial mice, AnkB and Caspr1 immunostaining was lost/abnormal compared with uninjured control ( Supplemental Figure 4 ) at anti-sulfatide antibody–exposed paranodes.…”

Section: Resultsmentioning

confidence: 99%

“…This was generated by immunising ganglioside-deficient mice with ganglioside liposomes or ganglioside-mimicking Campylobacter jejuni lipo-oligosaccharide, as described previously (22). Mouse monoclonal IgG3 anti-sulfatide antibody was also used, and generated as previously described (27). The following antibodies were used for immunostaining studies to identify proteins and complement: mouse anti-AnkB (UC Davis/NIH NeuroMab Facility Cat# N105/17, RRID:AB_2877200; 1:300) , mouse anti-AnkG (Thermo Fisher Scientific Cat# 33-8800, RRID:AB_2533145; 1:100); rabbit anti-Caspr1 (gifted by Professor Peles, Weizmann Institute, Rehovot, Israel; 1:1000); FITC-labelled rabbit anti-C3c (Agilent Cat# Q036805, RRID:AB_11180931; 1:300); rabbit anti-gliomedin (Abcam Cat# ab24483, RRID:AB_2111616; 1:100); mouse anti-human C5b-9 (Agilent Cat# M0777, RRID:AB_2067162; 1:50); rabbit anti-Kv1.1 Alomone Labs Cat# APC-009, RRID:AB_2040144; 1:200); rat anti-MBP (Bio-Rad Cat# MCA409S, RRID:AB_325004; 1:500); mouse anti-pan Nav (pNav; Sigma-Aldrich #8809; RRID:AB_477552; 1:100); rabbit anti-Nav1.6 (Sigma-Aldrich Cat# S0438, RRID:AB_477480; 1:100); rabbit anti-pan neurofascin (anti-panNFasc; gifted from Professor Brophy, University of Edinburgh, Edinburgh, UK; 1:1000); mouse anti-phosphorylated neurofilament-H antibody (NFH, BioLegend #801602 clone SMI31; RRID:AB_2715851; 1:1500).…”

Section: Antibodies and Reagents The Majority Of Experiments Used The...mentioning

confidence: 99%

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Schwann cell nodal membrane disruption triggers bystander axonal degeneration in a Guillain-Barré syndrome mouse model

McGonigal¹,

Barrie²,

Yao³

et al. 2022

Journal of Clinical Investigation

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In Guillain-Barré syndrome (GBS), both axonal and demyelinating variants can be mediated by complement-fixing anti-GM1 ganglioside autoantibodies that target peripheral nerve axonal and Schwann cell (SC) membranes, respectively. Critically, the extent of axon degeneration in both variants dictates long-term outcome. The differing pathomechanisms underlying direct axonal injury and the secondary "bystander" axonal degeneration following SC injury are unresolved. To investigate this, we generated glycosyltransferase-disrupted transgenic mice that express GM1 ganglioside either exclusively in neurons (GalNAcT -/--Tg(neuronal)) or glia (GalNAcT -/--Tg(glial)), thereby allowing anti-GM1 antibodies to solely target GM1 in either axonal or SC membranes, respectively. Myelinated axon integrity in distal motor nerves was studied in transgenic mice exposed to anti-GM1 antibody and complement in ex vivo and in vivo injury paradigms. Axonal targeting induced catastrophic acute axonal disruption as expected. When mice with GM1 in SC membranes were targeted, acute disruption of perisynaptic glia, and SC membranes at nodes of Ranvier (NoR) occurred. Following glial injury, axon disruption at nodes also developed sub-acutely, progressing to secondary axon degeneration. These models differentiate the distinctly different axonopathic pathways under axonal and glial membrane targeting conditions, and provide insights into primary and secondary axon injury, currently a major unsolved area in GBS research.

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Section: Resultsmentioning

confidence: 99%

Section: Antibodies and Reagents The Majority Of Experiments Used The...mentioning

confidence: 99%

Schwann cell nodal membrane disruption triggers bystander axonal degeneration in a Guillain-Barré syndrome mouse model

McGonigal¹,

Barrie²,

Yao³

et al. 2022

Journal of Clinical Investigation

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“…Anti-GM1 ganglioside antibody (DG2) and anti-GD1b antibody (MOG1) were used at 20 μg/ml. A new monoclonal anti-sulfatide antibody (GAMEG3) derived from mice inoculated with sulfatide-bearing liposomes was used to detect sulfatide (Meehan et al, 2018). The following primary antibodies were used: mouse anti-phosphorylated neurofilament-H antibody (NF-H, BioLegend; RRID:AB_2715851; 1:2000); rat anti-MBP (Bio-Rad; RRID:AB_325004; 1:500); mouse anti-pan Nav (Sigma-Aldrich; RRID:AB_477552; 1:100); mouse anti-ankyrin G (Thermo Fisher Scientific; RRID:AB_2533145; 1:100); rabbit anti-Caspr (gifted from Professor Peles, Weizmann Institute, Israel; 1:1500); rabbit anti-Kv1.1 (Alomone Laboratories; RRID:AB_2040144; 1:200); rabbit anti-Nav1.6 (Sigma-Aldrich; RRID:AB_477480; 1:100); rabbit anti-pan neurofascin (anti-pNFasc; gifted from Professor Brophy, University of Edinburgh, UK; 1:1000); and mouse anti-MAG antibody (gifted from Professor Brophy, University of Edinburgh, UK; 1:100).…”

Section: Methodsmentioning

confidence: 99%

Glial Sulfatides and Neuronal Complex Gangliosides Are Functionally Interdependent in Maintaining Myelinating Axon Integrity

et al. 2018

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Sulfatides and gangliosides are raft-associated glycolipids essential for maintaining myelinated nerve integrity. Mice deficient in sulfatide (cerebroside sulfotransferase knock-out, CST−/−) or complex gangliosides (β-1,4-N-acetylegalactosaminyltransferase1 knock-out, GalNAc-T−/−) display prominent disorganization of proteins at the node of Ranvier (NoR) in early life and age-dependent neurodegeneration. Loss of neuronal rather than glial complex gangliosides underpins the GalNAc-T−/− phenotype, as shown by neuron- or glial-specific rescue, whereas sulfatide is principally expressed and functional in glial membranes. The similarities in NoR phenotype of CST−/−, GalNAc-T−/−, and axo–glial protein-deficient mice suggests that these glycolipids stabilize membrane proteins including neurofascin155 (NF155) and myelin-associated glycoprotein (MAG) at axo–glial junctions. To assess the functional interactions between sulfatide and gangliosides, CST−/− and GalNAc-T−/− genotypes were interbred. CST−/−× GalNAc-T−/− mice develop normally to postnatal day 10 (P10), but all die between P20 and P25, coinciding with peak myelination. Ultrastructural, immunohistological, and biochemical analysis of either sex revealed widespread axonal degeneration and disruption to the axo–glial junction at the NoR. In addition to sulfatide-dependent loss of NF155, CST−/− × GalNAc-T−/− mice exhibited a major reduction in MAG protein levels in CNS myelin compared with WT and single-lipid-deficient mice. The CST−/− × GalNAc-T−/− phenotype was fully restored to that of CST−/− mice by neuron-specific expression of complex gangliosides, but not by their glial-specific expression nor by the global expression of a-series gangliosides. These data indicate that sulfatide and complex b-series gangliosides on the glial and neuronal membranes, respectively, act in concert to promote NF155 and MAG in maintaining the stable axo–glial interactions essential for normal nerve function.SIGNIFICANCE STATEMENT Sulfatides and complex gangliosides are membrane glycolipids with important roles in maintaining nervous system integrity. Node of Ranvier maintenance in particular requires stable compartmentalization of multiple membrane proteins. The axo–glial adhesion molecules neurofascin155 (NF155) and myelin-associated glycoprotein (MAG) require membrane microdomains containing either sulfatides or complex gangliosides to localize and function effectively. The cooperative roles of these microdomains and associated proteins are unknown. Here, we show vital interdependent roles for sulfatides and complex gangliosides because double (but not single) deficiency causes a rapidly lethal phenotype at an early age. These findings suggest that sulfatides and complex gangliosides on opposing axo–glial membranes are responsible for essential tethering of the axo–glial junction proteins NF155 and MAG, which interact to maintain the nodal complex.

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“…Anti‐sulphatide and anti‐ganglioside antibody binding patterns at the peripheral nerve node of Ranvier. (a) This panel has been modified from Figure 4(a), Meehan et al (2018) ( https://doi.org/10.1016/j.jneuroim.2018.07.004) with permission under the Creative Commons CC‐BY licence ( http://creativecommons.org/licenses/by/4.0/). Anti‐sulphatide antibody binding along the myelin and at the paranodal loops (green, *) was observed in the peripheral nerves (magenta) of wild‐type mice, while no binding was detected in CST −/− mice, thus demonstrating the specificity of the antibody to sulphatide.…”

Section: Gangliosides and Other Glycolipids At The Node Of Ranviermentioning

confidence: 99%

“…Indeed, in the absence of sulphatide, loss of NF155 immunostaining suggests a role in tethering this key paranodal axo‐glial adhesion molecule to the glial membrane (Hoshi et al, 2007 ; McGonigal et al, 2019 ). However, only recently, owing to the development of new anti‐sulphatide antibodies, have immunolabelling studies shown that sulphatide appears highly enriched in paranodal loops, as well as being present along the Schwann cell membrane (Meehan et al, 2018 ) (Figure 3a ).…”

Section: Gangliosides and Other Glycolipids At The Node Of Ranviermentioning

confidence: 99%

The role of gangliosides in the organisation of the node of Ranvier examined in glycosyltransferase transgenic mice

McGonigal

Willison

2021

Journal of Anatomy

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Gangliosides are sialic acid containing glycosphingolipids widely expressed in vertebrate plasma membranes and intracellular compartments (Ledeen & Wu, 2011;Sandhoff & Harzer, 2013;Yu et al., 2011). Whilst ganglioside biosynthesis and distribution is widespread throughout the body, the nervous system in particular is very highly enriched in complex gangliosides relative to other organs and tissues. Gangliosides are present in both central and peripheral nervous system (CNS and PNS) grey and white matter, where they have roles in development and homeostasis, modulating cell-cell recognition, signal transduction, growth and motility. Gangliosides are key components of lipid raft membrane domains where they have been widely studied (Simons & Toomre, 2000;Sonnino et al., 2015). In this review, we will focus our attention on the role of gangliosides in the maintenance of the mammalian node of Ranvier (NoR), a specialised membrane domain formed by myelin-forming cells: Schwann cells in the PNS and oligodendrocytes in the CNS (Figure 1). The NoR is a critical relay station in the electrophysiological functioning of myelinating nerve fibres, essential for saltatory conduction, promoted by high density clustering of voltage-gated sodium (Nav) channels on the nodal axolemma.Anchoring of the myelin paranodal loops by adhesion molecules and their associated cytoskeletal linker proteins in both the glial and axonal membranes at the paranode segregate and condense the ion channels (Pinatel & Faivre-Sarrailh, 2021;Rasband & Peles, 2021). Of note to our review, disruption to the expression of the paranodal axo-glial membrane protein complex composed of glial Neurofascin 155 (NF155), axonal contactin-associated

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Differential binding patterns of anti-sulfatide antibodies to glial membranes

Cited by 8 publications

References 50 publications

Schwann cell nodal membrane disruption triggers bystander axonal degeneration in a Guillain-Barré syndrome mouse model

Schwann cell nodal membrane disruption triggers bystander axonal degeneration in a Guillain-Barré syndrome mouse model

Glial Sulfatides and Neuronal Complex Gangliosides Are Functionally Interdependent in Maintaining Myelinating Axon Integrity

The role of gangliosides in the organisation of the node of Ranvier examined in glycosyltransferase transgenic mice

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