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Transcript profiling of different types of multiple sclerosis lesions yields FGF1 as a promoter of remyelination.

Mohan H, Friese A, Albrecht S, Krumbholz M, Elliott CL, Arthur A, Menon R, Farina C, Junker A, Stadelmann C, Barnett SC, Huitinga I, Wekerle H, Hohlfeld R, Lassmann H, Kuhlmann T, Linington C, Meinl E - Acta Neuropathol Commun (2014)

Bottom Line: We dissected remyelinated, demyelinated active, and demyelinated inactive white matter MS lesions, and compared transcript levels of myelination and inflammation-related genes using quantitative PCR on customized TaqMan Low Density Arrays.In remyelinated lesions, fibroblast growth factor (FGF) 1 was the most abundant of all analyzed myelination-regulating factors, showed a trend towards higher expression as compared to demyelinated lesions and was significantly higher than in control white matter.Two MS tissue blocks comprised lesions with adjacent de- and remyelinated areas and FGF1 expression was higher in the remyelinated rim compared to the demyelinated lesion core.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Neuroimmunology, Ludwig Maximilian University Munich, Marchioninistraße 15, D-81377, Munich, Germany. hema.mohan@ukmuenster.de.

ABSTRACT
Chronic demyelination is a pathological hallmark of multiple sclerosis (MS). Only a minority of MS lesions remyelinates completely. Enhancing remyelination is, therefore, a major aim of future MS therapies. Here we took a novel approach to identify factors that may inhibit or support endogenous remyelination in MS. We dissected remyelinated, demyelinated active, and demyelinated inactive white matter MS lesions, and compared transcript levels of myelination and inflammation-related genes using quantitative PCR on customized TaqMan Low Density Arrays. In remyelinated lesions, fibroblast growth factor (FGF) 1 was the most abundant of all analyzed myelination-regulating factors, showed a trend towards higher expression as compared to demyelinated lesions and was significantly higher than in control white matter. Two MS tissue blocks comprised lesions with adjacent de- and remyelinated areas and FGF1 expression was higher in the remyelinated rim compared to the demyelinated lesion core. In functional experiments, FGF1 accelerated developmental myelination in dissociated mixed cultures and promoted remyelination in slice cultures, whereas it decelerated differentiation of purified primary oligodendrocytes, suggesting that promotion of remyelination by FGF1 is based on an indirect mechanism. The analysis of human astrocyte responses to FGF1 by genome wide expression profiling showed that FGF1 induced the expression of the chemokine CXCL8 and leukemia inhibitory factor, two factors implicated in recruitment of oligodendrocytes and promotion of remyelination. Together, this study presents a transcript profiling of remyelinated MS lesions and identified FGF1 as a promoter of remyelination. Modulation of FGF family members might improve myelin repair in MS.

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FGF1 enhances remyelination in organotypic cerebellar slice cultures. After toxic demyelination, cerebellar slice cultures were allowed to remyelinate for 14 days in the absence or presence of 100 ng/ml FGF1. Myelination was assessed by immunostaining (a, b) and quantitative PCR (c–e). (a)Mpb (red), NFL (green) scale bar = 50 μm (b) Quantification of Mpb+/NFL+ area ratio in FGF1 treated slices compared to untreated controls. Mpb+ myelin formation is promoted by FGF1. Students t-test *P = 0.0341. (c-e) After 14 days RNA was extracted, cDNA obtained and transcript levels of (c)Mpb, (d)Mag and (e)Mog were measured by qPCR. One-way ANOVA **P < 0.01; *P < 0.05; All error bars represent SEM from three independent experiments.
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Fig6: FGF1 enhances remyelination in organotypic cerebellar slice cultures. After toxic demyelination, cerebellar slice cultures were allowed to remyelinate for 14 days in the absence or presence of 100 ng/ml FGF1. Myelination was assessed by immunostaining (a, b) and quantitative PCR (c–e). (a)Mpb (red), NFL (green) scale bar = 50 μm (b) Quantification of Mpb+/NFL+ area ratio in FGF1 treated slices compared to untreated controls. Mpb+ myelin formation is promoted by FGF1. Students t-test *P = 0.0341. (c-e) After 14 days RNA was extracted, cDNA obtained and transcript levels of (c)Mpb, (d)Mag and (e)Mog were measured by qPCR. One-way ANOVA **P < 0.01; *P < 0.05; All error bars represent SEM from three independent experiments.

Mentions: To determine whether FGF1 would also promote remyelination, we investigated its effects on demyelinated organotypic cerebellar cultures derived from newborn mouse pups. Slice cultures myelinated in vitro in 12 days, they were then demyelinated using lysolecithin and allowed to recover in the presence or absence of FGF1. Analysis of cultures 14 days later revealed that FGF1 promoted remyelination; myelin basic protein (MBP) immunoreactivity increased approximately 1.5-fold as assessed by immunofluorescence microscopy (Figure 6a and 6b). The MBP expression was normalized based on the axonal density as measured by NFL staining. This increase in MBP positive immunoreactivity was paralleled by an increase of Mbp transcripts (Figure 6c). Further, we observed an increase in transcript level of Mag (Figure 6d), but not consistently of Mog (Figure 6e).Figure 6


Transcript profiling of different types of multiple sclerosis lesions yields FGF1 as a promoter of remyelination.

Mohan H, Friese A, Albrecht S, Krumbholz M, Elliott CL, Arthur A, Menon R, Farina C, Junker A, Stadelmann C, Barnett SC, Huitinga I, Wekerle H, Hohlfeld R, Lassmann H, Kuhlmann T, Linington C, Meinl E - Acta Neuropathol Commun (2014)

FGF1 enhances remyelination in organotypic cerebellar slice cultures. After toxic demyelination, cerebellar slice cultures were allowed to remyelinate for 14 days in the absence or presence of 100 ng/ml FGF1. Myelination was assessed by immunostaining (a, b) and quantitative PCR (c–e). (a)Mpb (red), NFL (green) scale bar = 50 μm (b) Quantification of Mpb+/NFL+ area ratio in FGF1 treated slices compared to untreated controls. Mpb+ myelin formation is promoted by FGF1. Students t-test *P = 0.0341. (c-e) After 14 days RNA was extracted, cDNA obtained and transcript levels of (c)Mpb, (d)Mag and (e)Mog were measured by qPCR. One-way ANOVA **P < 0.01; *P < 0.05; All error bars represent SEM from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359505&req=5

Fig6: FGF1 enhances remyelination in organotypic cerebellar slice cultures. After toxic demyelination, cerebellar slice cultures were allowed to remyelinate for 14 days in the absence or presence of 100 ng/ml FGF1. Myelination was assessed by immunostaining (a, b) and quantitative PCR (c–e). (a)Mpb (red), NFL (green) scale bar = 50 μm (b) Quantification of Mpb+/NFL+ area ratio in FGF1 treated slices compared to untreated controls. Mpb+ myelin formation is promoted by FGF1. Students t-test *P = 0.0341. (c-e) After 14 days RNA was extracted, cDNA obtained and transcript levels of (c)Mpb, (d)Mag and (e)Mog were measured by qPCR. One-way ANOVA **P < 0.01; *P < 0.05; All error bars represent SEM from three independent experiments.
Mentions: To determine whether FGF1 would also promote remyelination, we investigated its effects on demyelinated organotypic cerebellar cultures derived from newborn mouse pups. Slice cultures myelinated in vitro in 12 days, they were then demyelinated using lysolecithin and allowed to recover in the presence or absence of FGF1. Analysis of cultures 14 days later revealed that FGF1 promoted remyelination; myelin basic protein (MBP) immunoreactivity increased approximately 1.5-fold as assessed by immunofluorescence microscopy (Figure 6a and 6b). The MBP expression was normalized based on the axonal density as measured by NFL staining. This increase in MBP positive immunoreactivity was paralleled by an increase of Mbp transcripts (Figure 6c). Further, we observed an increase in transcript level of Mag (Figure 6d), but not consistently of Mog (Figure 6e).Figure 6

Bottom Line: We dissected remyelinated, demyelinated active, and demyelinated inactive white matter MS lesions, and compared transcript levels of myelination and inflammation-related genes using quantitative PCR on customized TaqMan Low Density Arrays.In remyelinated lesions, fibroblast growth factor (FGF) 1 was the most abundant of all analyzed myelination-regulating factors, showed a trend towards higher expression as compared to demyelinated lesions and was significantly higher than in control white matter.Two MS tissue blocks comprised lesions with adjacent de- and remyelinated areas and FGF1 expression was higher in the remyelinated rim compared to the demyelinated lesion core.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Neuroimmunology, Ludwig Maximilian University Munich, Marchioninistraße 15, D-81377, Munich, Germany. hema.mohan@ukmuenster.de.

ABSTRACT
Chronic demyelination is a pathological hallmark of multiple sclerosis (MS). Only a minority of MS lesions remyelinates completely. Enhancing remyelination is, therefore, a major aim of future MS therapies. Here we took a novel approach to identify factors that may inhibit or support endogenous remyelination in MS. We dissected remyelinated, demyelinated active, and demyelinated inactive white matter MS lesions, and compared transcript levels of myelination and inflammation-related genes using quantitative PCR on customized TaqMan Low Density Arrays. In remyelinated lesions, fibroblast growth factor (FGF) 1 was the most abundant of all analyzed myelination-regulating factors, showed a trend towards higher expression as compared to demyelinated lesions and was significantly higher than in control white matter. Two MS tissue blocks comprised lesions with adjacent de- and remyelinated areas and FGF1 expression was higher in the remyelinated rim compared to the demyelinated lesion core. In functional experiments, FGF1 accelerated developmental myelination in dissociated mixed cultures and promoted remyelination in slice cultures, whereas it decelerated differentiation of purified primary oligodendrocytes, suggesting that promotion of remyelination by FGF1 is based on an indirect mechanism. The analysis of human astrocyte responses to FGF1 by genome wide expression profiling showed that FGF1 induced the expression of the chemokine CXCL8 and leukemia inhibitory factor, two factors implicated in recruitment of oligodendrocytes and promotion of remyelination. Together, this study presents a transcript profiling of remyelinated MS lesions and identified FGF1 as a promoter of remyelination. Modulation of FGF family members might improve myelin repair in MS.

Show MeSH
Related in: MedlinePlus