Images were captured from stained frozen sections using a fluorescence microscope equipped with 4objectives

Images were captured from stained frozen sections using a fluorescence microscope equipped with 4objectives. demyelinated hippocampus. Our results suggest that demyelination might lead to the impairment of neuronal transport, but not cause increased level of hyperphosphorylated tau and beta amyloid. Our research demonstrates remyelination might be an effective pathway to recover the function of neuronal axons and cognition in MS. and 0.05). Demyelination in the hippocampus In the hippocampal cortex, we detected the myelin-associated proteins, including MBP, 2,3-cyclic nucleotide 3-phosphodiesterase (CNPase), and proteolipid protein (PLP). We found that the expression Cgp 52432 of MBP was significantly declined in the cuprizine-fed mice compared with the control group and it was increased in the LINGO-1 antibody treated group (Physique ?(Figure2A).2A). However, other myelin-associated proteins, including CNPase and PLP, were reduced but not significantly in the cuprizine-fed mice (Physique 2B and 2C). In immunofluorescence staining, we also found decreased expression of MBP in subregions of the hippocampal cortex, Cgp 52432 including the cornu ammonis 1 (Ca1), Ca3 and dentate gyrus (DG), in the cuprizine-fed mice, and LINGO-1 antibody could partly increase the level of MBP (Physique 2D-2L). Open in a separate window Physique 2 Demyelination in the hippocampusMyelin associated proteins in the hippocampus, including MBP A., CNPase B. and PLP C., were detected among the three groups using Western-blot. Figures D.-L. were displayed the immunofluorescence staining of MBP in subregions of the hippocampus, including Cgp 52432 Ca3 (D-F), Ca1 (G-I) and DG (J-L). *denotes statistical significance compared with controls ( 0.05). #denotes statistical significance compared with the cuprizine-fed mice without treatment ( 0.05). Images were captured from stained frozen sections using a fluorescence microscope equipped with 10objectives. Level bar, 1000m. Reduction of proteins essential for axonal transport in the hippocampus Efficient axonal transport is essential to maintain neuronal function [32]. KLC is responsible for binding of cargo during fast anterograde transport, whereas dynein (Dyn) is the important proteins involve in retrograde transport [32]. We found that the level of KLC was significantly declined in the hippocampus of the cuprizine-fed mice and it was slightly increased, but not significantly, after LINGO-1 antibody treatment (Physique ?(Figure3A).3A). However, no significant switch of Dyn level was measured among the three groups of mice (Physique ?(Figure3B).3B). In immunofluorescence staining, we also found decreased expression of KLC in subregions of the demyelinated hippocampal cortex, including Ca3 and Ca1, and LINGO-1 antibody could partly increase the level of KLC (Physique 3C-3H). However, the expression of KLC was comparable in DG among the three groups (Physique 3I-3K). Open in a separate window Physique 3 Reduction of proteins essential for axonal transport in the hippocampusA. KLC expression in the hippocampus was compared among the three groups. B. Dyn expression in the hippocampus was compared among the three groups. Figures C.-K. were displayed the immunofluorescence staining of KLC in subregions of the hippocampus, including Ca3 (C-E), Ca1 (F-H) and DG (I-K).*denotes Cgp 52432 statistical significance compared with controls ( 0.05). Images were captured from stained frozen sections using a fluorescence microscope equipped with 40objectives. Level bar, 200m. The neurofilaments (NFs) are a major component of the neuronal cytoskeleton, involved in providing structural support for the axon and regulating axon diameter, essential for the formation of axonal networks [33]. In the hippocampal cortex, the expression of NF200 in the cuprizine-fed mice was lower than that in the control mice, and after six-week LINGO-1 antibody treatment, NF200 was increased significantly (Physique ?(Figure4A).4A). Also NFL was significantly reduced in the cuprizine-fed mice and was increased but Bivalirudin Trifluoroacetate not significantly (Physique ?(Figure4B)4B) after six-week LINGO-1 antibody treatment. In immunofluorescence staining, we Cgp 52432 also found decreased expression of NF200 in subregions of the demyelinated hippocampal cortex, including Ca3, Ca1 and DG, and LINGO-1 antibody could increase the.