Histone H1 improves regeneration after mouse spinal cord injury and changes shape and gene expression of cultured astrocytes

Article type: Research Article

Authors: Kleene, Ralf^{a; 2} | Loers, Gabriele^{a; 2} | Jakovcevski, Igor^a | Mishra, Bibhudatta^{a; 1} | Schachner, Melitta^{b; c; *}

Affiliations: [a] Zentrum für Molekulare Neurobiologie, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany | [b] Department of Cell Biology and Neuroscience, Keck Center for Collaborative Neuroscience, Rutgers University, Piscataway, NJ, USA | [c] Center for Neuroscience, Shantou University Medical College, Shantou, Guangdong, China

Correspondence: [*] Corresponding author: Melitta Schachner, Center for Neuroscience, Shantou University Medical College, 22 Xin Ling Road, Shantou, Guangdong 515041, China. Tel.: +86 754 8890 0276; Fax: +86 754 8890 0236; E-mails: [email protected] or [email protected].

Note: [1] Present address: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Note: [2] These authors contributed equally.

Abstract: Background:We have shown that histone H1 is a binding partner for polysialic acid (PSA) and that it improves functional recovery, axon regrowth/sprouting, and target reinnervation after mouse femoral nerve injury. Objective:Here, we analyzed whether histone H1 affects functional recovery, axon regrowth/sprouting, and target reinnervation after spinal cord injury of adult mice. Furthermore, we tested in vitro histone H1’s effect on astrocytic gene expression, cell shape and migration as well as on cell survival of cultured motoneurons. Methods:We applied histone H1 to compressed spinal cord and determined functional recovery and number of fibrillary acidic protein (GFAP)- and neuron-glial antigen 2 (NG2)- positive glial cells, which contribute to glial scarring. Histone H1’s effect on migration of astrocytes, astrocytic gene expression and motoneuronal survival was determined using scratch-wounded astroglial monolayer cultures, astrocyte cultures for microarray analysis, and motoneuron cell culture under oxidative stress conditions, respectively. Results:Histone H1 application improves locomotor functions and enhances monoaminergic and cholinergic reinnervation of the spinal cord. Expression levels of GFAP and NG2 around the lesion site were decreased in histone H1-treated mice relative to vehicle-treated mice six weeks after injury. Histone H1 reduced astrocytic migration, changed the shape of GFAP- and NG2-positive glial cells and altered gene expression. Gene ontology enrichment analysis indicated that in particular genes coding for proteins involved in proliferation, differentiation, migration and apoptosis are dysregulated. The up- and down-regulation of distinct genes was confirmed by qPCR and Western blot analysis. Moreover, histone H1 reduced hydrogen peroxide-induced cell death of cultured motoneurons. Conclusions:The combined observations indicate that histone H1 locally applied to the lesion site, improves regeneration after spinal cord injury. Some of these beneficial functions of histone H1 in vivo and in vitro can be attributed to its interaction with PSA-carrying neural cell adhesion molecule.

Keywords: Glial scar, histone H1, polysialic acid, mouse spinal cord injury, synaptic plasticity, regeneration, locomotion, axonal regrowth, astrogliosis, migration

DOI: 10.3233/RNN-190903

Journal: Restorative Neurology and Neuroscience, vol. 37, no. 4, pp. 291-313, 2019