Purification of Oligodendrocyte Precursor Cells from Rat Cortices by Immunopanning

Dugas, Jason C.; Emery, Ben

doi:10.1101/pdb.prot070862

Cited by 53 publications

(62 citation statements)

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“…Unless otherwise indicated, OPCs were purified from enzymatically dissociated P7–P8 Sprague-Dawley (Charles Rivers) rat brains by immunopanning and grown in serum-free defined medium, as described previously (Dugas and Emery, 2013). Mouse OPCs were purified from transgenic mice by immunopanning as described (Emery and Dugas, 2013).…”

Section: Methodsmentioning

confidence: 99%

CNS Myelin Wrapping Is Driven by Actin Disassembly

et al. 2015

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SUMMARY Myelin is essential in vertebrates for the rapid propagation of action potentials, but the molecular mechanisms driving its formation remain largely unknown. Here we show that the initial stage of process extension and axon ensheathment by oligodendrocytes requires dynamic actin filament assembly by the Arp2/3 complex. Unexpectedly, subsequent myelin wrapping coincides with the upregulation of actin disassembly proteins, rapid disassembly of the oligodendrocyte actin cytoskeleton, and does not require Arp2/3. Inducing loss of actin filaments drives oligodendrocyte membrane spreading and myelin wrapping in vivo, and the actin disassembly factor gelsolin is required for normal wrapping. We show that myelin basic protein, a protein essential for CNS myelin wrapping whose role has been unclear, is required for actin disassembly, and its loss phenocopies loss of actin disassembly proteins. Together these findings provide insight into the molecular mechanism of myelin wrapping and identify it as an actin-independent form of mammalian cell motility.

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Section: Methodsmentioning

confidence: 99%

CNS Myelin Wrapping Is Driven by Actin Disassembly

et al. 2015

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“…Rat cortical OPCs were isolated with immunopanning as previously described (Dugas and Emery, 2013) with the following modifications. Cerebral cortices from P1-7 Sprague-Dawley rats were dissected and then digested in papain (Worthington) in DPBS at 35°C for 30-45 minutes.…”

Section: Star Methodsmentioning

confidence: 99%

Fibrinogen Activates BMP Signaling in Oligodendrocyte Progenitor Cells and Inhibits Remyelination after Vascular Damage

Petersen

Ryu

Chang

et al. 2017

Neuron

148

153

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Summary Blood-brain barrier (BBB) disruption alters the composition of the brain microenvironment by allowing blood proteins into the CNS. However, whether blood-derived molecules serve as extrinsic inhibitors of remyelination is unknown. Here we show that the coagulation factor fibrinogen activates the bone morphogenetic protein (BMP) signaling pathway in oligodendrocyte progenitor cells (OPCs) and suppresses remyelination. Fibrinogen induces phosphorylation of Smad 1/5/8 and inhibits OPC differentiation into myelinating oligodendrocytes (OLs), while promoting an astrocytic fate in vitro. Fibrinogen effects are rescued by BMP type I receptor inhibition using dorsomorphin homologue 1 (DMH1) or CRISPR/Cas9 activin A receptor type I (ACVR1) knockout in OPCs. Fibrinogen and the BMP target Id2 are increased in demyelinated multiple sclerosis (MS) lesions. Therapeutic depletion of fibrinogen decreases BMP signaling and enhances remyelination in vivo. Targeting fibrinogen may be an upstream therapeutic strategy to promote the regenerative potential of CNS progenitors in diseases with remyelination failure.

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“…Oligodendrocyte precursor cells (OPCs) were purified from rat pups of P7–P9 by immunopanning (19). Primary OPCs and CG4 cells (20) were kept in a proliferative condition by supplementing the Sato media (19) with PDGF (10 μg/mL), NT3 (1 μg/mL), and CNTF (10 μg/mL). Primary OPCs and CG4 cells were maintained in a humidified 8% CO 2 incubator at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Homo-trimerization is essential for the transcription factor function of Myrf for oligodendrocyte differentiation

Kim

Choi

Fan

et al. 2017

Nucleic Acids Research

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Myrf is a key transcription factor for oligodendrocyte differentiation and central nervous system myelination. We and others have previously shown that Myrf is generated as a membrane protein in the endoplasmic reticulum (ER), and that it undergoes auto-processing to release its N-terminal fragment from the ER, which enters the nucleus to work as a transcription factor. These previous studies allow a glimpse into the unusual complexity behind the biogenesis and function of the transcription factor domain of Myrf. Here, we report that Myrf N-terminal fragments assemble into stable homo-trimers before ER release. Consequently, Myrf N-terminal fragments are released from the ER only as homo-trimers. Our re-analysis of a previous genetic screening result in Caenorhabditis elegans shows that homo-trimerization is essential for the biological functions of Myrf N-terminal fragment, and that the region adjacent to the DNA-binding domain is pivotal to its homo-trimerization. Further, our computational analysis uncovered a novel homo-trimeric DNA motif that mediates the homo-trimeric DNA binding of Myrf N-terminal fragments. Importantly, we found that homo-trimerization defines the DNA binding specificity of Myrf N-terminal fragments. In sum, our study elucidates the molecular mechanism governing the biogenesis and function of Myrf N-terminal fragments and its physiological significance.

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Purification of Oligodendrocyte Precursor Cells from Rat Cortices by Immunopanning

Cited by 53 publications

References 0 publications

CNS Myelin Wrapping Is Driven by Actin Disassembly

CNS Myelin Wrapping Is Driven by Actin Disassembly

Fibrinogen Activates BMP Signaling in Oligodendrocyte Progenitor Cells and Inhibits Remyelination after Vascular Damage

Homo-trimerization is essential for the transcription factor function of Myrf for oligodendrocyte differentiation

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