Anti-LINGO-1 improved remyelination and neurobehavioral deficit in cuprizone-induced demyelination

Abstract

Objectives: Central nervous system demyelination is the main feature of multiple sclerosis (MS). The most important unmet need in MS is use of treatments that delay the progression of the disease. Leucine-rich repeat and Immunoglobulin-like domain containing NOGO receptor-interacting protein 1(LINGO-1) have been known as inhibitors of oligodendrocyte differentiation and myelination.

Materials and methods: We investigated LINGO-1 antibody effects on remyelination and neurobehavioral deficit using cuprizone-induced demyelination. Animals were randomly divided into three groups (n = 10): (1) Control group; received the regular diet, (2) CPZ group; normal saline was injected intraperitoneally, and (3) Treatment group; LINGO-1 antibody (10 mg/kg) was injected IP once every six days for 3 weeks. We assessed the level of myelin basic protein (MBP), neurofilament heavy chain (NF200), and Brain-derived neuroprotective factor (BDNF) in the corpus callosum (CC) by immunostaining against MBP, NF200, and BDNF.

Results: We found decreased levels of MBP, NF200, and BDNF in demyelinated CC, and anti-LINGO-1 treatment improved demyelinated structures. Furthermore, motor impairment was measured by Open-field (OFT) and Balance beam tests. In the treatment group, motor impairment was significantly improved.

Conclusion: These results provide evidence that LINGO-1 antibody can improve remyelination and neurobehavioral deficit.

Keywords: Brain-derived neurotrophic – factor; Cuprizone; Multiple sclerosis; Myelin basic protein; Nogo receptor 1; Remyelination.

Figure 1

Schematic representation of the experimental protocols. The control group was fed normal chow. The cuprizone group was fed a 0.2% cuprizone (CPZ) diet for 6 weeks without return to normal chow. Behavioral assessments were conducted in the beginning, at three-weeks, and the end of the experimental period, and brain tissue was collected for IHF assessment

Figure 2

Determining the myelination levels by staining against myelin basic protein (MBP) as a myelin marker in the corpus callosum (CC). (A) Immunofluorescence staining against MBP to evaluate the myelination level in CC. (B) Quantitative analysis of the MBP-stained sections shows the protective effect of the LINGO-1 antibody. Data were shown as Mean±SEM. MBP was increased in the treatment group in comparison with the control group (P<0.001). Also, there was a significant decrease in MBP in the CPZ+fed group in comparison with the control group (P<0.001). One-way ANOVA was used for statistical analysis followed by Tukey's post-hoc test

Figure 3

Effect of LINGO-1 antibody on NF200 level in the corpus callosum (CC). (A) The level of NF200 in CC (B) Quantitative analysis of NF200 in sections of CC compared between the three groups. Data were shown as Mean±SEM. NF200 level was increased in the treatment group (P<0.001) and decreased in the CPZ+fed group in comparison with the control group (P<0.001). One-way ANOVA was used as statistical analysis followed by Tukey's post-hoc test. Data presented as mean ± SEM

Figure 4

Effect of Anti-LINGO 1 on BDNF. IHF staining against BDNF (A) was performed to investigate the effect of LINGO-1 antibody on BDNF in CC for control, cuprizone, and cuprizone+LINGO-1 groups. (B) Quantitative analysis of BDNF in sections. Data were shown as Mean±SEM. BDNF level was increased in the treatment group (P<0.001) and decreased in the CPZ+fed group in comparison with the control group (P<0.001). One-way ANOVA was used for statistical analysis followed by Tukey's post-hoc test. Data presented as mean±SEM

Figure 5

Schematic representation of mice's balance beam test. Values represent mean ±SEM