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SNAT - RNA sequencing of murine developing peripheral nerves

ABSTRACT: SNAT - RNA sequencing of murine developing peripheral nerves

PROVIDER: PRJNA573696 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10164714.fastq.gz	Fastqsanger.gz
	SRR10164715.fastq.gz	Fastqsanger.gz
	SRR10164716.fastq.gz	Fastqsanger.gz
	SRR10164717.fastq.gz	Fastqsanger.gz
	SRR10164718.fastq.gz	Fastqsanger.gz

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SNAT - RNA sequencing of murine developing peripheral nerves

Project description:RNA sequencing of peripheral nerve lysates covering various developmental stages in mice (E13.5, E17.5, P1, P5, P14, P24, P60)

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ACTL6a knockout peripheral nerve RNA seq at postnatal day 3

Project description:Cells need to integrate chemical and physical signals into transcriptional programs. In the peripheral nerves, axonal caliber selection by specialized Schwann cells, is critical for developmental myelination. However, only mechanisms through which Schwann cells sense chemical signals are well characterized. We identify ACtin-Like protein 6a (ACTL6a), a component of the SWI/SNF chromatin remodeling complex, as critical for axonal caliber recognition and developmental myelination. ACTL6a is expressed in the developing nerve and, when activated by contact with axons or nanofibers of specific caliber, it promotes the eviction of repressive histone marks thereby enhancing the transcriptional program of myelination. Mutant mice with Schwann cells lacking ACTL6a display aberrant recognition of axonal caliber, resulting in defective radial sorting and lower levels of transcripts regulating myelination of developing nerves. We suggest that developing peripheral nerves require an ACTL6a-dependent integration of physical and chemical signals in Schwann cells to release of repressive histone modifications and promote myelination.

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SNAT - Single cell RNA sequencing of mouse cells sorted from sciatic nerves at postnatal day 1 (P1) and P60

Project description:SNAT - Single cell RNA sequencing of mouse cells sorted from sciatic nerves at postnatal day 1 (P1) and P60

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The molecular profile and ability to promote axon regeneration after peripheral nerve injury by Repair Schwann cells is distinctly different from that for developing Schwann cells

Project description:Since Schwann cells (SCs) support axonal growth at development as well as after peripheral nerve injury (PNI), developing SCs might be able to promote axon regeneration after PNI. The purpose of the current study was to elucidate the capability of developing SCs to induce axon regeneration after PNI. SC precursors (SCPs), immature SCs (ISCs), repair SCs (RSCs) from injured nerves, and non-RSCs from intact nerves were tested by grafting into acellular region of rat sciatic nerve with crush injury. Both of developing SCs completely failed to support axon regeneration, whereas both of mature SCs, especially RSCs, induced axon regeneration. Further, RSCs but not SCPs promoted neurite outgrowth of adult dorsal root ganglion neurons. Transcriptome analysis revealed that the gene expression profiles were distinctly different between RSCs and SCPs. These findings indicate that developing SCs are markedly different from mature SCs in terms of functional and molecular aspects and that RSC is a viable candidate for regenerative cell therapy for PNI.

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Transcriptional profiling of mouse peripheral nerves to the single-cell level to build a sciatic nerve ATlas (SNAT).

Project description:Peripheral nerves are organ-like structures containing diverse cell types to optimize function. This interactive assembly includes mostly axon-associated Schwann cells, but also endothelial cells of supporting blood vessels, immune system-associated cells, barrier-forming cells of the perineurium surrounding and protecting nerve fascicles, and connective tissue-resident cells within the intra-fascicular endoneurium and inter-fascicular epineurium. We have established transcriptional profiles of mouse sciatic nerve-inhabitant cells to foster the fundamental understanding of peripheral nerves. To achieve this goal, we have combined bulk RNA sequencing of developing sciatic nerves up to the adult with focused bulk and single-cell RNA sequencing of Schwann cells throughout postnatal development, extended by single-cell transcriptome analysis of the full sciatic nerve both perinatally and in the adult. The results were merged in the transcriptome resource Sciatic Nerve ATlas (SNAT: https://www.snat.ethz.ch). We anticipate that insights gained from our multi-layered analysis will serve as valuable interactive reference point to guide future studies.

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Project description:Peripheral innervation plays an important role in regulating tissue repair and regeneration. Here, we provide evidence that injured peripheral nerves provide a reservoir of mesenchymal precursor cells that can directly contribute to murine digit tip regeneration and skin repair. In particular, using single-cell RNA sequencing and lineage tracing we identify transcriptionally-distinct mesenchymal cell populations within the control and injured adult nerve, including neural crest-derived cells in the endoneurium with characteristics of mesenchymal precursor cells. Culture and transplantation studies show that these nerve-derived mesenchymal cells have the potential to differentiate into non-nerve lineages. Moreover, following digit tip amputation, the neural crest-derived nerve mesenchymal cells contribute to the regenerative blastema and ultimately to the regenerated bone. Similarly, neural crest derived nerve mesenchymal cells contribute to the dermis during skin wound healing. These findings support a model where peripheral nerves directly contribute mesenchymal precursor cells to promote repair and regeneration of injured mammalian tissues.

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Project description:Immune cells infiltrate peripheral nerves after injury and with autoimmunity, but their net effect is divergent. After injury, immune cells are reparative, while in autoimmune neuropathies (e.g., GBS and CIDP), immune cells are pro-inflammatory and promote autoimmune demyelination. An understanding of immune cell phenotypes that distinguish these conditions may therefore reveal new therapeutic targets for switching immune cells from an inflammatory to a reparative state. In an autoimmune neuropathy mouse model, we used single cell transcriptomic studies of sciatic nerves to discover a TNFα signaling axis induced by IFNγ. Macrophages were the main source of TNFα, and autocrine TNFα signaling upregulated multiple autoimmunity-associated genes (e.g., Clec4e; Marcksl1; Cxcl1; Cxcl10). TNFα signaling was absent in nerve macrophages after injury, suggesting that TNFα signaling defines autoimmunity-associated pro-inflammatory macrophages. Finally, TNFα/TNFα signaling blockade ameliorated PNS autoimmunity development, which provides pre-clinical evidence that the TNFα axis may be effectively targeted to resolve inflammatory neuropathies.

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