Abstract
Oligodendrocytes produce the myelin that is critical for rapid neuronal transmission in the central nervous system (CNS). Disruption of myelin has devastating effects on CNS function, as in the demyelinating disease multiple sclerosis (MS). Microglia are the endogenous immune cells of the CNS and play a central role in demyelination and repair. There is a need for new potential therapies that regulate myelination and microglia to promote repair. Agathisflavone (FAB) is a non-toxic flavonoid that is known for its anti-inflammatory and neuroprotective properties. Here, we examined the effects of FAB (5-50 μM) on myelination and microglia in organotypic cerebellar slices prepared from P10-P12 Sox10-EGFP and Plp1-DsRed transgenic mice. Immunofluorescence labeling for myelin basic protein (MBP) and neurofilament (NF) demonstrates that FAB significantly increased the proportion of MBP + /NF + axons but did not affect the overall number of oligodendroglia or axons, or the expression of oligodendroglial proteins CNPase and MBP. FAB is known to be a phytoestrogen, but blockade of α- or β- estrogen receptors (ER) indicated the myelination promoting effects of FAB were not mediated by ER. Examination of microglial responses by Iba1 immunohistochemistry demonstrated that FAB markedly altered microglial morphology, characterized by smaller somata and reduced branching of their processes, consistent with a decreased state of activation, and increased Iba1 protein expression. The results provide evidence that FAB increases the extent of axonal coverage by MBP immunopositive oligodendroglial processes and has a modulatory effect upon microglial cells, which are important therapeutic strategies in multiple neuropathologies.
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Funding
This work was supported by the Multiple Sclerosis Society (AB, FP) and the Biotechnology and Biological Sciences Research Council (AB, grant number BB/M029379/1). We thank the Coordination of Personnel Improvement of Higher Level (CAPES, Process PDSE Nº 88881.133939/2016–01; and Process PGCI Nº 88881.117666/2016–01), the National Council for Scientific and Technological Development (CNPq, Scholarship Nº 140333/2016–9, Post-Graduation Program in Immunology- Federal University of Bahia-Brazil), Foundation for Research Support of the State of Bahia (FAPESB, Process Nº INT 0016/2016) and Post-Graduation Program in Immunology- Federal University of Bahia-Brazil for fellowship (CAPES Process Nº 0001). This work was additionally funded by MS Society of Canada (3573) and Canadian Institutes for Health Research (CIHR PS 166120) operating grants as well as Canada Research Chair Tier II award in Neural Stem Cell Biology awarded to A.V.
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MMAA performed all experimentation, analyzed, interpreted the data and wrote manuscript. TF assisted with western blot analysis. FP helped to perform immunohistochemistry and confocal microscope images and revised it critically for intellectual content. JMD and JPD performed the chemical analysis and extraction of agathisflavone. VDAS and CSS revised it critically for intellectual content. AMB, SLC, AV supervised the study, edited and reviewed the manuscript. All authors read and approved the final manuscript.
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AMB is a share-holder in the company ‘GliaGenesis Ltd.’. Otherwise, the authors report no conflicts of interest, including personal or financial.
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de Almeida, M.M.A., Pieropan, F., Footz, T. et al. Agathisflavone Modifies Microglial Activation State and Myelination in Organotypic Cerebellar Slices Culture. J Neuroimmune Pharmacol 17, 206–217 (2022). https://doi.org/10.1007/s11481-021-09991-6
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DOI: https://doi.org/10.1007/s11481-021-09991-6