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| Status |
Public on Sep 29, 2023 |
| Title |
Genetically perturbed myelin as a risk factor for neuroinflammation-driven axon degeneration |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Axon degeneration and neurological dysfunction in myelin diseases is often attributed to loss of myelin. Perturbed myelinating glia can instigate chronic neuroinflammation and contribute to demyelination and axonal damage. We have previously shown in mice that distinct defects in the proteolipid protein 1 gene result in axonal damage which is largely driven by cytotoxic T cells targeting myelinating oligodendrocytes. Here we show in these mutants that persistent ensheathment with perturbed myelin poses a risk for axon degeneration, neuron loss, and behavioral decline. We demonstrate that CD8+ T cell-driven axonal damage is less likely to progress towards degeneration when axons are efficiently demyelinated by activated microglia. Mechanistically, we show that cytotoxic T cell effector molecules induce aberrant cytoskeletal plasticity within myelinating glial processes and constriction of axons at paranodal domains. Our study identifies detrimental axon-glia interactions which promote neurodegeneration and possible therapeutic targets for disorders associated with myelin defects and neuroinflammation.
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| Overall design |
Around 15,000 CD45lowSiglec-H+ and CD45-O1+ single cells were sorted per sample using a FACSAria III (BD Biosciences) before being encapsulated into droplets with the Chromium Controller (10x Genomics) and processed according to the manufacturer’s specifications. Briefly, every transcript captured in all the cells encapsulated with a bead was uniquely barcoded using a combination of a 16-base pair (bp) 10x barcode and a 10-bp unique molecular identifier (UMI). Complementary DNA libraries ready for sequencing on Illumina platforms were generated using the Chromium Single Cell 3′ Library & Gel Bead Kit v2 (10x Genomics) according to the detailed protocol provided by the manufacturer. Libraries were quantified by Qubit 3.0 Fluorometer (Thermo Fisher Scientific) and quality was checked using a 2100 Bioanalyzer with High Sensitivity DNA kit (Agilent Technologies). Libraries were pooled and sequenced with a NovaSeq 6000 platform (S1 Cartridge; Illumina) in paired-end mode to reach a mean of 44,131 reads per single cell. A total of 13,556, 9,955, and 12,191 cells were captured and a median gene number per cell of 2,268, 2,562, and 2,165 could be retrieved for adult Wt, PLPmut, and PLPtg cells, respectively.
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| Contributor(s) |
Groh J, Abdelwahab T, Kattimani Y, Hörner M, Loserth S, Gudi V, Adalbert R, Imdahl F, Saliba A, Coleman M, Stangel M, Martini R |
| Citation(s) |
37903797 |
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| Submission date |
Jan 30, 2023 |
| Last update date |
Nov 16, 2023 |
| Contact name |
Fabian Imdahl |
| E-mail(s) |
fabian.imdahl@helmholtz-hiri.de
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| Phone |
09313188471
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| Organization name |
helmholtz institute for RNA based infection research
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| Department |
Single-cell-center
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| Street address |
Josef-Schneider Str. 2
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| City |
Würzburg |
| State/province |
Bavaria |
| ZIP/Postal code |
97080 |
| Country |
Germany |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA929453 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE224032_RAW.tar |
274.9 Mb |
(http)(custom) |
TAR (of MTX, TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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