A channelopathy contributes to cerebellar dysfunction in a model of multiple sclerosis
Shannon D. Shields PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorXiaoyang Cheng PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorAndreas Gasser PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorCarl Y. Saab PhD
Department of Surgery, Rhode Island Hospital, Brown Alpert Medical School, Providence, RI
Department of Neuroscience, Brown University, Providence, RI
Search for more papers by this authorLynda Tyrrell MS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorEmmanuella M. Eastman BS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorMasashi Iwata PhD
Department of Surgery, Rhode Island Hospital, Brown Alpert Medical School, Providence, RI
Department of Neuroscience, Brown University, Providence, RI
Search for more papers by this authorPamela J. Zwinger BS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorJoel A. Black PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorSulayman D. Dib-Hajj PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorCorresponding Author
Stephen G. Waxman MD, PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Neuroscience and Regeneration Research Center, Veterans' Affairs Connecticut Healthcare System, 950 Campbell Ave, Bldg 34, West Haven, CT 06516Search for more papers by this authorShannon D. Shields PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorXiaoyang Cheng PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorAndreas Gasser PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorCarl Y. Saab PhD
Department of Surgery, Rhode Island Hospital, Brown Alpert Medical School, Providence, RI
Department of Neuroscience, Brown University, Providence, RI
Search for more papers by this authorLynda Tyrrell MS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorEmmanuella M. Eastman BS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorMasashi Iwata PhD
Department of Surgery, Rhode Island Hospital, Brown Alpert Medical School, Providence, RI
Department of Neuroscience, Brown University, Providence, RI
Search for more papers by this authorPamela J. Zwinger BS
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorJoel A. Black PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorSulayman D. Dib-Hajj PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Search for more papers by this authorCorresponding Author
Stephen G. Waxman MD, PhD
Department of Neurology and Center for Neuroscience and Regeneration Research, Yale University School of Medicine, New Haven, CT
Rehabilitation Research Center, Veterans Affairs Connecticut Healthcare System, West Haven, CT
Neuroscience and Regeneration Research Center, Veterans' Affairs Connecticut Healthcare System, 950 Campbell Ave, Bldg 34, West Haven, CT 06516Search for more papers by this authorAbstract
Objective:
Cerebellar dysfunction in multiple sclerosis (MS) contributes significantly to disability, is relatively refractory to symptomatic therapy, and often progresses despite treatment with disease-modifying agents. We previously observed that sodium channel Nav1.8, whose expression is normally restricted to the peripheral nervous system, is present in cerebellar Purkinje neurons in a mouse model of MS (experimental autoimmune encephalomyelitis [EAE]) and in humans with MS. Here, we tested the hypothesis that upregulation of Nav1.8 in cerebellum in MS and EAE has functional consequences contributing to symptom burden.
Methods:
Electrophysiology and behavioral assessment were performed in a new transgenic mouse model overexpressing Nav1.8 in Purkinje neurons. We also measured EAE symptom progression in mice lacking Nav1.8 compared to wild-type littermates. Finally, we administered the Nav1.8-selective blocker A803467 in the context of previously established EAE to determine reversibility of MS-like deficits.
Results:
We report that, in the context of an otherwise healthy nervous system, ectopic expression of Nav1.8 in Purkinje neurons alters their electrophysiological properties, and disrupts coordinated motor behaviors. Additionally, we show that Nav1.8 expression contributes to symptom development in EAE. Finally, we demonstrate that abnormal patterns of Purkinje neuron firing and MS-like deficits in EAE can be partially reversed by pharmacotherapy using a Nav1.8-selective blocker.
Interpretation:
Our results add to the evidence that a channelopathy contributes to cerebellar dysfunction in MS. Our data suggest that Nav1.8-specific blockers, when available for humans, merit study in MS. Ann Neurol 2012;71:186–194
Supporting Information
Additional Supporting Information can be found in the online version of this article.
| Filename | Description |
|---|---|
| ANA_22665_sm_SuppFig1.tif1.6 MB | Supplementary Figure 1. Confirmation of impaired coordinated motor behaviors in a second, independently derived line of L7-1.8TG mice excludes insertion-site effects. Black bars, L7-1.8TG mice; white bars, WT littermates. A. Transgenic mice perform poorly in a wire hang test (p=0.0015, n= 20 WT, 18 TG). B. Transgenic mice fall with shorter latency from a constant speed rotarod (p=0.0287, n=18 WT, 10 TG). C. Grip strength is similar for wildtype and transgenic mice (p=0.2004, n= 13 WT, 9 TG). D. Response to heat stimuli in the hindpaw radiant heat (Hargreaves) test is similar for wildtype and transgenic mice (p=0.9432, n=8 WT, 8 TG). E. Response to mechanical stimuli in the von Frey test is similar for wildtype and transgenic mice (p=0.561, n=8 WT, 8 TG). |
| ANA_22665_sm_SuppFig2.tif9 MB | Supplementary Figure 2. Selective upregulation of Nav1.8 in Purkinje neurons and not in other regions of the central nervous system. A. Full sagittal brain section from a wildtype mouse with EAE, showing in situ hybridization signal for Nav1.8. Note that the only specific labeling is present in the Purkinje cell layer of the cerebellum (arrowheads). Inset, higher magnification view of boxed region. B. Full sagittal brain section from a Nav1.8KO mouse with EAE, showing in situ hybridization signal for Nav1.8. No labeling is present anywhere in the brain, although shadowing in the white matter tracts is similar to wildtype. Asterisk denotes a fold in the tissue. Inset, higher magnification view of boxed region. C. Spinal cord section from a Nav1.8-Cre-reporter mouse without EAE. Note that, although dense fluorescent reporter labeling is present in primary afferent terminals in the dorsal horn, no intrinsic spinal cord neurons express Nav1.8. D. Spinal cord section from a Nav1.8-Cre-reporter mouse on Day 15 after induction of EAE. Primary afferent terminals are labeled as in the control mouse, and no new expression of Nav1.8 is induced in any spinal cord region. |
| ANA_22665_sm_SuppFig3.tif2.5 MB | Supplementary Figure 3. Immune cell infiltration is similar in WT and Nav1.8KO mice with EAE. A. Top panel, No CD45-positive cells were found in lumbar spinal cord samples from control mice without EAE. Middle panel, CD45-positive immune cell infiltration was prominent in spinal cord samples from WT mice on day 10 after induction of EAE. Bottom panel, CD45-positive immune cell infiltration was prominent in spinal cord samples from Nav1.8KO mice at the same stage of EAE. B. CD45-positive immune cell infiltration was similar in WT and Nav1.8KO mice (p>0.05, n=4/group). |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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