B01 - Function of satellite oligodendrocytes in remyelination

https://rdp.sfb274.de/ (Research Data Platform) https://gepris.dfg.de/gepris/projekt/408885537 (DFG Gepris entry) The central nervous system (CNS) is a terminally differentiated tissue, where any insult carries a heightened risk - yet the tissue response to these insults is variable and can range from irreversible destruction to almost complete recovery. The rules that instruct these divergent outcomes are still unknown. The aim of this CRC is therefore to understand the biology of the multicellular ...

As tissue-resident macrophages of the central nervous system parenchyma, microglia perform diverse essential functions during homeostasis and perturbations1. They primarily interact with neurons by means of synaptic engulfment and through the rapid elimination of apoptotic cells and non-functional synapses2. Here, by combining unbiased lipidomics and high-resolution spatial lipid imaging, deep single-cell transcriptome analysis and novel cell-type-specific mutants, we identified a previously ...

Compartmentalized inflammation is a key driver of multiple sclerosis (MS) progression, but the mechanisms sustaining its persistence remain unclear. A hallmark of this persistent and slowly evolving inflammatory process is chronic active MS lesions. We generated a high-resolution, single-cell molecular and spatial atlas of such lesions by combining single-nucleus RNA sequencing (snRNA-seq) with multiplexed error-robust fluorescence in situ hybridization (MERFISH). Within lesion rims, we identified ...

Remyelination requires innate immune system function, but how exactly microglia and macrophages clear myelin debris after injury and tailor a specific regenerative response is unclear. Here, we asked whether pro-inflammatory microglial/macrophage activation is required for this process. We established a novel toxin-based spinal cord model of de- and remyelination in zebrafish and showed that pro-inflammatory NF-κB–dependent activation in phagocytes occurs rapidly after myelin injury. We found ...

CNS nuclei were isolated from frozen thalamic specimen, stained with anti-NeuN and anti-Olig2, FACS purified (DAPI+NeuN-Olig2-), and analyzed with snRNA-seq

Brains were homogenized, stained, microglia were FACS-purified, RNA was isolated, cDNA was produced and sequencing libraries were prepared.

CNS nuclei were isolated from frozen thalamic specimen, FACS purified, and analyzed with snRNA-seq

CNS nuclei were isolated from frozen thalamic specimen, FACS purified, and analyzed with snRNA-seq

CNS nuclei were isolated from frozen CNS tissue specimen, stained with anti-NeuN and anti-Olig2, FACS purified (DAPI+NeuN-Olig2-), and analyzed with snRNA-seq

Frozen autopsy brain tissue from fourteen multiple-sclerosis donors and three neurologically healthy controls was sectioned at –20 °C into 80–100 µm slices, and an 8 mm punch (~25 mg) of white matter or matched control tissue was collected from each region of interest . Nuclei were liberated with the 10x Genomics Chromium Nuclei Isolation Kit, using an extended lysis–debris-removal protocol tailored to lipid-rich white matter; the final suspension was stained with propidium iodide and counted ...

In this study, our experimental design focused on the detailed analysis of lesion areas in the brain tissues of patients with multiple sclerosis (MS). First, we collected brain tissue samples from 14 MS patients. Using specialized staining techniques, we accurately identified the locations of MS lesions within the brain tissues. Subsequently, we used a brain puncher to extract the brain tissue regions containing lesions from each sample. These precisely cut samples were then analyzed on the ...

8-week-old male Cx3cr1CreERT2/+; Abca1flox/flox; Abcg1flox/flox experimental animals (KO) and Cx3cr1CreERT2/+ controls (Ctrl) were used for EAE experiments. Myelin oligodendrocyte glycoprotein (MOG)-EAE was induced in mice purchased from Charles River Laboratories by subcutaneous immunization with 200 mg of MOG peptide 35–55 (MOG35–55) in complete Freund’s adjuvant (Mycobacterium tuberculosis at 3.75 mg ml−1; BD) and intraperitoneal injection twice with 500 ng of pertussis toxin (Sigma). Animals ...