(a) Experimental strategy of conditional Fasn allele inactivation upon Olig2-driven Cre expression in vivo. (b) Schematic of thoracic spinal cord. Square insert ‘c’: White matter (WM) where …
(a) Representative immunostaining of cross-sectioned thoracic spinal cord white matter from P14 CT and cMU mice, n = 5 mice for each, CT and cMU. Cre-dependent expression of the reporter Rosa26-loxPs…
(a) Schematic of the thoracic spinal cord. Square insert ‘b’: Gray matter (GM) area where subsequent immunostaining images were acquired. WM = white matter. (b) Representative immunostaining of …
(a) Representative immunostaining of ventral white matter in cross-sectioned spinal cords of P4 and P10 mice identifying proliferating (EdU+) OLs (Olig2+; examples indicated by arrowheads), n = 3 …
(a) Representative immunostaining of ventral white matter in cross-sections of lumbar spinal cords from P14 CT mice depicting marginal (if any) FASN expression in OPCs (PDGFRα+; examples indicated …
(a) Representative EM images of P14 control (CT, a’ and a’’) and conditional mutant (cMU, b’ and b’’) white matter from the ventral funiculi of lumbar spinal cords. cMUs show more naked axons, …
(a) Representative immunostaining of cross-sectioned optic nerves from P14 CT and cMU mice, n = 5 mice for each, CT and cMU. Cre-dependent expression of the reporter Rosa26-loxPstoploxP-YFP allele …
(a) Representative EM images of optic nerves from P14 control (CT) and conditional mutant (cMU) mice. Note the increased number of axons not-yet enwrapped by myelin in cMUs when compared to CTs. …
(a) Representative EM images of corpus callosum from P180 CT and cMU mice. cMUs show slightly more axons not-yet enwrapped by myelin compared to CTs. cMU axons are also covered by thinner myelin …
(a) Exemplary EM images of aberrant myelin-axon profiles showing vacuolation (examples indicated by arrowheads) in the ventral white matter of spinal cords of P14 cMUs. Where identifiable, axons …
(a) EM images of aberrant myelinated axon profiles (examples indicated by arrowheads) in the optic nerve of P14 cMU mice. Scale bars: I = 2 µm, II = 1 µm, III = 2 µm, IV = 1 µm, V = 1 µm, VI = 1 µm. …
Lipids extracted from myelin isolated from spinal cords of control (CT) and mutant (cMU) mice were identified and quantified by liquid chromatography-mass spectrometry. Individual species of 10 …
(a, b) GeneOntology analysis depicting biological processes affected by down-regulated (a) and up-regulated (b) transcripts in P14 optic nerves of mutant (cMU) mice. Metacore Pathway analysis …
Expressed transcripts in optic nerves from control and cMU mice at P14.
Graph of qRT-PCR analysis of Sox10, Myrf and MBP in optic nerves of P14 CT and cMU mice. Data are normalized to β-actin and to the mean of CTs (unpaired two-tailed two sample Student’s t-test; …
(a) Scheme of the experimental design. (b) Exemplary EM micrographs of the ventral funiculus of the lumbar spinal cord from mutant (cMU) and control (CT) mice that were fed a standard (STD) or a …
(a) Tamoxifen-mediated nuclear translocation of Pdgfrα-driven CreERT2 induces conditional Fasn allele inactivation (i-cMU) and expression of yellow fluorescent protein (YFP). (b) Timeline of …
(a) Exemplary immunostaining of cross-sectioned thoracic spinal cord white matter from 5 weeks post tamoxifen, n = 3 CT mice. Cre-dependent expression of the reporter Rosa26-loxPstoploxP-YFP allele …
(a) Representative immunostaining of lesions of control (CT) and inducible conditional mutant (i-cMU) mice on cross sections of ventral white matter of the spinal cord at 7, 10, 14 and 21 days …
(a) Representative immunostaining of the lesion area in cross sections of the spinal cord of induced control (CT) mice at 7 days post-lysolecithin injection (dpl). Images depict marginal expression …
(a) Representative immunostaining in the lesion area of cross-sectioned thoracic spinal cords of control (CT) and inducible conditional mutant (i-cMU) mice at 7 dpl. n = 5 mice for each, CT and …
(a) Representative immunostaining of the lesion area in cross sections of the thoracic spinal cord at 12 days post-lysolecithin injection (dpl) from control (CT) and inducible conditional mutant …
(a) Representative EM images of lesions in the spinal cord of control (CT) and inducible conditional mutant (i-cMU) mice during remyelination at 14, 21 and 56 days post-lysolecithin injection (dpl). …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (M. musculus) | Fasnlox/lox | PMID:16054078 | MGI:3765070 | Dr. Clay F. Semenkovich, Washington University, St. Louis USA |
Genetic reagent (M. musculus) | PdgfraCreERT2 | PMID:18849983 | MGI:3832569 | Dr. William D. Richardson, University College London, London UK |
Genetic reagent (M. musculus) | Olig2Cre | Jackson Laboratory | Stock#:011103; MGI:3810299 | PMID:18691547 |
Genetic reagent (M. musculus) | Rosa26-loxPstoploxP-YFP | Jackson Laboratory | Stock#:006148; MGI:3621481 | PMID:11299042 |
Sequence-based reagent | Genotyping primer: Fasn forward | PMID:29434029 | 5'-GGATAGCTGTGTAGTGTAACCAT-3’ | |
Sequence-based reagent | Genotyping primer: Fasn reverse | PMID:29434029 | 5’-GGTCACCCAGCAGGAAAGGGC- 3’ | |
Sequence-based reagent | Genotyping primer: Cre forward | PMID: 29434029 | 5’-TTCCCGCAGAACCTGAAGATGTTCG-3’ | |
Sequence-based reagent | Genotyping primer: Cre reverse | PMID: 29434029 | 5’-GGGTGTTATAAGCAATCCCCAGAAATG-3’ | |
Sequence-based reagent | Genotyping primer: Rosa26-loxPstoploxP-YFP forward | PMID: 28522536 | 5’-AAAGTCGCTCTGAGTTGTTAT-3’ | |
Sequence-based reagent | Genotyping primer: Rosa26-loxPstoploxP-YFP reverse transgenic | PMID: 28522536 | 5’-GCGAAGAGTTTGTCCTCAACC-3’ | |
Sequence-based reagent | Genotyping primer: Rosa26-loxPstoploxP-YFP reverse wildtype | PMID: 28522536 | 5’-GGAGCGGGAGAAATGGATATG-3’ | |
Sequence- based reagent | qRT-PCR primer: SOX10 Forward | This paper | 5’-CCGACCAGTACCCTCACCT-3’ | |
Sequence-based reagent | qRT-PCR primer: SOX10 Reverse | This paper | 5’- TCAATGAAGGGGCGCTTGT-3’ | |
Sequence-based reagent | qRT-PCR primer: MYRF Forward | This paper | 5’-ATGGAGGTGGTGGACGAGAC-3’ | |
Sequence-based reagent | qRT-PCR primer:MYRF Reverse | This paper | 5’-GGCGTCCTCTTTGCCAATGT-3’ | |
Sequence- based reagent | qRT-PCR primer: MBP Forward | This paper | 5’-ACACGAGAACTACCCATTATGGC-3’ | |
Sequence- based reagent | qRT-PCR primer: MBP Reverse | This paper | 5’-CCAGCTAAATCTGCTGAGGGGA-3’ | |
Sequence- based reagent | qRT-PCR primer: Actin Forward | PMID: 28880149 | 5’-GTCCACACCCGCCACC-3’ | |
Sequence- based reagent | qRT-PCR primer: Actin Reverse | PMID: 28880149 | 5’-GGCCTCGTCACCCACATAG-3’ | |
Antibody | Rabbit polyclonal anti-cleaved Caspase 3 | Cell signaling Technology, Danvers, MA, USA | Cat# 9661; RRID:AB_2341188 | 1:500 dilution |
Antibody | Mouse monoclonal anti-CC1 | Merck Millipore, Billerica, MA, USA | Cat; # OP80; RRID:AB_2057371 | Development: 1:200 dilution; Remyelination 1:300 dilution |
Antibody | rabbit polyclonal anti-FASN | Abcam, UK | Cat# Ab22759; RRID:AB_732316 | 1:200 dilution |
Antibody | Chicken polyclonal anti-GFP | Abcam, UK | Cat# Ab13970; RRID:AB_300798 | 1:1000 dilution |
Antibody | mouse monoclonal anti-ki67 | Dako Agilent, Santa Clara, CA, USA | Cat# m7249; clone MIB-5; RRID:AB_2250503 | 1:200 dilution |
Antibody | rat monoclonal anti-MBP | Serotec/BioRad Laboratories, Hercules, CA, USA | Cat# MCA409S; RRID:AB_325004 | Development: 1:200 dilution; Remyelination 1:300 dilution |
Antibody | Goat polyclonal anti-Olig2 | R and D Systems, Minneapolis, MN, USA | Cat# AF2418; RRID:AB_2157554 | 1:25 dilution |
Antibody | mouse monoclonal anti-Olig2 | Merck Millipore, Billerica, MA, USA | Cat# MABN50; clone 211F1.1; RRID:AB_10807410 | 1:1000 dilution |
Antibody | rabbit polyclonal anti-Olig2 | Merck Millipore, Billerica, MA, USA | Cat# AB9610; RRID:AB_570666 | Development: 1:500 dilution; Remyelination 1:400 dilution |
Antibody | Rabbit monoclonal anti-PDGFRα | Cell signaling Technology, Danvers, MA, USA | Cat# 3174; RRID:AB_2162345 | 1:500 dilution |
Commercial assay or kit | Click-iT EdU Assay | Thermo Fisher Scientific, Waltham, MA, USA | Cat# C10337 | |
Commercial assay or kit | Mouse blocking reagent | Vector Laboratories, Burlingame, CA, USA | Cat# MKB-2213 | |
Commercial assay or kit | Streptavidin/Biotin blocking Kit | Vector Laboratories, Burlingame, CA, USA | Cat# SP-2002 | |
Commercial assay or kit | Qiagen MiniKit (RNeasy Mini Kit) | Qiagen, Hilden, Germany | Cat# 74104 | |
Commercial assay or kit | TruSeq Stranded mRNA Sample Prep Kit | Illumina, San Diego, CA, USA | Cat# 20020594 | |
Commercial assay or kit | Maxima RT-Kit | Thermo Fisher Scientific, Waltham, MA, USA | Cat# K1641 | |
Chemical compound, drug | Lysolecithin | Sigma-Aldrich, Sant Louis, MO, USA | Cat# L4129 | |
Chemical compound, drug | Tamoxifen | Sigma-Aldrich, Sant Louis, MO, USA | Cat# T5648 | |
Chemical compound, drug | Lipid standards | Avanti Polar Lipids, Alabaster, AL, USA | ||
Software, algorithm | Lipid Data Analyzer software | PMID: 29058722 | ||
Software, algorithm | Photoshop CS5 or CS6 | Adobe | ||
Software, algorithm | FIJI | ImageJ (http://imagej.nih.gov/ij/) | ||
Software, algorithm | STAR Aligner(v2.5.1b) | PMID: 23104886 | ||
Software, algorithm | RSEM (v1.2.22) | PMID: 21816040 | ||
Software, algorithm | EdgeR | PMID: 19910308 | ||
Software, algorithm | Metacore (vs6.33) | Thomson Reuters | ||
Other | Standard diet (STD) | KLIBA NAFAG, Provimi KLIBA, Switzerland | Cat# 3437 | |
Other | High fat diet (HFD) | KLIBA NAFAG, Provimi KLIBA, Switzerland | Cat# 2127 |