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MAG alters the secretion of proteins involved in CNS plasticity

ABSTRACT: Myelin-associated glycoprotein (MAG) is a potent inhibitor of neurite growth in the central nervous system. Label free LC-MS2 quantitation was performed on the secretome from three sets of rat neuron cultures in two states: control, and exposed to MAG. Differential analysis of conditioned media revealed that MAG exposure up-regulates LTBP3, LTBP4, S100A6, septin-7 and neurofascin 186, while down-regulating frataxin, MAP6, syntenin-1 and GAP-43. When combined with other results from this study, these findings confirm that MAG induces secretion of transforming growth factor beta, which increases the expression of chondroitin sulfate proteoglycans after spinal cord injury. They also demonstrate that MAG has a generalized effect on the secretion of proteins that affect CNS plasticity - increasing the secretion of growth-inhibiting proteins, while suppressing the secretion of proteins that promote axonal growth. These are new insights into the mechanisms underlying MAG's ability to inhibit axonal regeneration.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Rattus Norvegicus (ncbitaxon:10116)

SUBMITTER: Sari Hannila

PROVIDER: MSV000089160 | MassIVE | Wed Mar 30 08:18:00 BST 2022

REPOSITORIES: MassIVE

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Project description:Spinal cord injury (SCI) often leads to persistent functional deficits due to severe neuron and glial loss, and to limited axonal regeneration after injury. Here we show that the transplantation of human dental stem cells into the completely transected adult rat spinal cord resulted in a significant recovery of hindlimb locomotor functions. These stem cells exhibited three major neuro-regenerative activities. First, they inhibited the SCI-induced apoptosis of neurons, astrocytes, and oligodendrocytes, improving the preservation of neuronal filaments and myelin sheaths. Second, they promoted the regeneration of transected axons by directly inhibiting multiple axon growth inhibitors, including chondroitin sulfate proteoglycan and myelin-associated glycoprotein, by paracrine mechanisms. Third, they replaced lost cells by differentiating into mature oligodendrocytes under the extreme conditions of SCI. Our data demonstrate that tooth-derived stem cells may provide novel therapeutic benefits for treating SCI through both cell-autonomous and paracrine neuro-regenerative activities. Human dental stem cells were isolated from exfoliated deciduous teeth extracted for clinical purposes were collected at Nagoya University School of Medicine, under approved guidelines set by Nagoya University (H-73, 2003). The ethics committee of Nagoya University approved the experimental protocols (permission number 8-2). Mesenchymal stem cells of human Bone marrow line were obtained from Lonza. Total RNAs were isolated and quantified by spectrophotometer. RNA integrity was checked on 1% agarose gels. RT reactions were carried out with Superscript III reverse transcriptase (Invitrogen) using 1 μg of total RNA in a 50 μl total reaction volume. Microarray experiments were carried out using a CodeLink™ Human Whole Genome Bioarray (Applied Microarrays, Inc. Tempe, AZ) at Filgen, Inc. (Nagoya, Japan). The arrays were scanned using a GenePix4000B Array Scanner (Molecular Devices, Sunnyvale, CA), and the data was analyzed by using MicroArray Data Analysis Tool Ver3.2 (Filgen, Inc.).

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MAG alters the secretion of proteins involved in CNS plasticity

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