Knockdown of Lingo1b protein promotes myelination and oligodendrocyte differentiation in zebrafish

Abstract

Demyelinating diseases include multiple sclerosis, which is a neurodegenerative disease characterized by immune attacks on the central nervous system (CNS), resulting in myelin sheath damage and axonal loss. Leucine-rich repeat and immunoglobulin domain-containing neurite outgrowth inhibitory protein (Nogo) receptor-interacting protein-1 (LINGO-1) have been identified as a negative regulator of oligodendrocytes differentiation. Targeted LINGO-1 inhibition promotes neuron survival, axon regeneration, oligodendrocyte differentiation, and remyelination in diverse animal models. Although studies in rodent models have extended our understanding of LINGO-1, its roles in neural development and myelination in zebrafish (Danio rerio) are not yet clear. In this study, we cloned the zebrafish homolog of the human LINGO-1 and found that lingo1b regulated myelination and oligodendrocyte differentiation. The expression of lingo1b started 1 (mRNA) and 2 (protein) days post-fertilization (dpf) in the CNS. Morpholino oligonucleotide knockdown of lingo1b resulted in developmental abnormalities, including less dark pigment, small eyes, and a curly spinal cord. The lack of lingo1b enhanced myelination and oligodendrocyte differentiation during embryogenesis. Furthermore, immunohistochemistry and movement analysis showed that lingo1b was involved in the axon development of primary motor neurons. These results suggested that Lingo1b protein functions as a negative regulator of myelination and oligodendrocyte differentiation during zebrafish development.

Keywords: 5-MIS; 5-bp mismatch control; ANOVA; Acc Dist; BDNF; CDS; CNS; Differentiation; EM; GAPDH; GFP; IPP; Ig; Image-Pro Plus; LINGO-1; LRR; LRR and Ig domain-containing Nogo receptor-interacting protein-1; Lingo1b; MAs; MBP; MOs; MS; MS-222; Mauthner axons; Multiple sclerosis; Myelination; NF1; NOGO; OKR; OPC; ORF; Olig2; Oligodendrocyte; PBS; PCR; PNS; Q-PCR; RNA-mediated interference; RNAi; SC; SDS; SE; SMART; SNP; STRING; Search Tool for the Retrieval of Interacting Genes/Proteins; Simple Modular Architecture Research Tool; TALENs; TBST; Tris-buffered saline-Tween 20; TrkB; UTR; WB; WISH; WT; ZFNs; Zebrafish; accumulation distance; analysis of variance; brain-derived neurotrophic factor; central nervous system; coding DNA sequence; day post-fertilization; dpf; electron microscopy; glyceraldehyde 3-phosphate dehydrogenase; green fluorescent protein; hours post-fertilization; hpf; immunoglobulin; leucine-rich repeat; morpholino oligonucleotides; multiple sclerosis; myelin basic protein; neurite outgrowth inhibitory protein; neurofibromin 1; oligodendrocyte lineage transcription factor 2; oligodendrocyte precursor cells; open reading frame; optokinetic response; pMNs; peripheral nervous system; phosphate buffered saline; polymerase chain reaction; primary motor neurons; real-time quantitative-polymerase chain reaction; single nucleotide polymorphism; sodium dodecyl sulfate; spinal cord; standard error; transcription activator-like effector nucleases; tricaine methane-sulfonate; tyrosine-related kinase B; untranslated region; western blots; whole-mount in situ hybridization; wild type; zinc finger nucleases.