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Axonal damage and demyelination in long-term dorsal root ganglia cultures from a rat model of Charcot–Marie–Tooth type 1A disease

Authors

  • Lucilla Nobbio,

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
    2. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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  • Gianfranco Gherardi,

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
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  • Tiziana Vigo,

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
    2. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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  • Mario Passalacqua,

    1. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
    2. Department of Experimental Medicine, Biochemistry Section, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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  • Edon Melloni,

    1. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
    2. Department of Experimental Medicine, Biochemistry Section, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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  • Michele Abbruzzese,

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
    2. Bio-imaging and Molecular Physiology Institute, CNR, Genoa, Italy
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  • Gianluigi Mancardi,

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
    2. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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  • Klaus-Armin Nave,

    1. Max-Planck-Institute of Experimental Medicine, Department of Neurogenetics, Hermann-Rein-Strasse 3, D-37075 Goettingen, Germany
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  • Angelo Schenone

    1. Department of Neurosciences, Ophthalmology and Genetics, University of Genoa, Italy, Via De Toni, 5, 16132, Genoa, Italy
    2. Center of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV, 16132, Genoa, Italy
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Dr Lucilla Nobbio, as 1above
E-mail: lnobbio@neurologia.unige.it

Abstract

Clinical progression in hereditary and acquired demyelinating disorders of both the central and peripheral nervous system is mainly due to a time-dependent axonal impairment. We established 90-day dorsal root ganglia (DRG) cultures from a rat model of Charcot–Marie–Tooth type 1A (CMT1A) neuropathy to evaluate the structure of myelin and axons, and the expression of myelin-related proteins and cytoskeletal components, by morphological and molecular techniques. Both wild-type and CMT1A cultures were rich in myelinated fibres. Affected cultures showed dysmyelinated internodes and focal myelin swellings. Furthermore, uncompacted myelin and smaller axons with increased neurofilament (NF) density were found by electron microscopy, and Western blots showed higher levels of nonphosphorylated NF. Confocal microscopy demonstrated an abnormal distribution of the myelin-associated glycoprotein which, instead of being expressed at the noncompact myelin level, showed focal accumulation along the internodes while other myelin proteins were normally distributed. These findings suggest that CMT1A DRG cultures, similarly to the animal model and human disease, undergo axonal atrophy over a period of time. This model may be utilized to study the molecular changes underlying demyelination and secondary axonal impairment. As axonal damage may occur after just 3 months and tissue cultures represent a strictly controlled environment, this model may be ideal for testing neuroprotective therapies.

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