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Whole cell response to receptor stimulation involves many deep and distributed subcellular biochemical processes

ABSTRACT: To identify subcellular processes that enable a whole cell response, we stimulated Neuro 2A cells with the cannabinoid receptor agonist HU210 that triggers neurite outgrowth. After 4 different stimulation periods we harvest the cells and subjected them to bulk RNAseq. Differentially expressed genes after each stimulation time were subjected to dynamic and standard enrichment analysis using the Molecular Biology of the Cell Ontology. We predicted the involvement of multiple canonical subcellular processes (SCPs) . Using siRNA ablation experiments of representative SCP genes we documented the involvement of the predicted SCPs in neurite outgrowth.

ORGANISM(S): Mus musculus

PROVIDER: GSE198622 | GEO | 2022/10/11

REPOSITORIES: GEO

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Metadata Entered in Submitting Proteomic Data to PRIDE

Project description:For the identification of potential pathways and their interactions that underlie a complex change in cellular state, such as neurite outgrowth, we stimulated the neuroblastoma cell line N2A with HU210, a potent activator of the cannabinoid receptor type I, followed by transcriptomic and proteomic analysis. Differentially expressed genes and proteins obtained at multiple timepoints were subjected to standard and dynamic enrichment analysis using the Molecular Biology of the Cell Ontology. Many of the identified pathways described constitutive cellular functions. Involvement of predicted pathways in neurite outgrowth was confirmed by siRNA knock-down of pathway representative genes in primary rat cortical neurons. Dynamic modeling of pathway activities suggested that the identified increase in vesicle membrane back-transported from the growth cone to the cell body ensures recycling of components needed by forward moving vesicle.

2022-10-14 | PXD036706 | Pride

Scalable generation of Schwann cell precursors from human pluripotent stem cells

Project description:Schwann cells (SCs) are an absolute prerequisite for development of effective treatment of myelin disorders and nerve injuries. However, human sources of functional, myelinating SCs are extremely limited. Here, we have developed a novel, efficient strategy for producing directly an unlimited supply of functional human SCs via successful derivation of expandable Schwann cell precursors (SCPs) from human pluripotent stem cells (hPSCs) (hPSC-SCPs). Functional and molecular characteristics of SCs from hPSC-SCPs (hPSC-SCP-SCs) appeared to be similar to those of authentic Schwann cells. As a novel therapeutic target, transplanted hPSC-SCP-SCs effectively promoted axonal regeneration through directly myelinating regenerated-axons in sciatic nerve injured mice. Here, we present hPSC-SCPs and hPSC-SCP-SCs as an outstanding resource to use for investigating human Schwann cell pathology and biology and for translational approaches to PNS and CNS repair and regeneration.

2017-08-15 | GSE95004 | GEO

Deletion of the gene family of small chlorophyll-binding proteins (ScpABCDE) offsets C/N homeostasis in synechocystis PCC 6803

2015-03-30 | GSE59371 | GEO

SCP-mediated chlorophyll synthesis and Photosystem II assembly maintains nitrogen homeostasis in Synechocystis sp. PCC 6803

Project description:Small CAB-like proteins (SCPs) are single-helix light-harvesting-like proteins found in all organisms performing oxygenic photosynthesis. We investigated the function of these stress-induced proteins in the cyanobacterium Synechocystis sp. PCC 6803 by comparing a strain, which expresses SCPs constitutively, with a mutant lacking all five SCPs. In the absence of SCPs, cells were larger, and showed irregular thylakoid structure and cell-surfaces. Deletion of scp genes strongly affected the carbon-nitrogen balance, causing accumulation of carbohydrates and a decrease in N-rich compounds (proteins and chlorophyll a). Data from transcriptomic and metabolomic experiments demonstrated that SCPs mediated stabilization of chlorophyll a under stress conditions is crucial to maintain the C/N balance. SCPs diminished the formation of reactive oxygen species, preventing cellular damage by adjusting the de-novo production of chlorophyll a to demand levels. Lack of SCP expression under stress conditions had a large impact on the metabolism of the entire cell. Synechocystis 6803 PSI-less (Shen et al 1993) and PSI-less/ScpABCDE- (Xu et al 2004) mutants were cultivated at 30°C at a light intensity of 4–5 μmol photons m–2 s–1 in BG-11 growth medium (Rippka et al 1979) supplemented with 10 mM glucose. Cells (40–50 ml) from both control and the mutant cultures were collected at mid-log phase by rapid filtration (Pall Supor 800 Filter, 0.8 mm) and total RNA extracted after Pinto et al. (Pinto et al 2009). Single-stranded cDNA preparation, labelling, and hybridization onto the microarray slides were performed after Eisenhut et al. (2007). The microarray slides were scanned with an Agilent Microarray Scanner (model G2505B) with default settings.

2015-03-30 | E-GEOD-59371 | biostudies-arrayexpress

Proteomic analysis to examine the functional consequences of intratumor heterogeneity

Project description:To examine the functional consequences of intratumor heterogeneity, subclonal populations (SCPs) were derived from the MDA-MB-468 triple-negative breast cancer cell line. Characterization of these SCPs revealed considerable variation in SCP tumor forming capacity with SCP #32 (SCP32) having an exceptional high tumor forming capacity. To investigate the proteomic differences between tumors of SCP32 and other SCPs, in TMT1, isobaric tandem mass tag (TMT) labeling combined with LC-MS/MS (TMT-MS) was performed on tumors of SCP03, SCP29 and SCP32 (n = 3 each) collected at 2 months post transplantation. In TMT2, isobaric tandem mass tag (TMT) labeling combined with LC-MS/MS (TMT-MS) was performed on tumors of the parental MDA-MB-468 cell line (n = 2), and both barcoded and non-barcoded versions SCP29 and SCP32 (two each) collected at 2 months post transplantation.

2022-02-16 | PXD022325 | Pride

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.

2022-01-13 | GSE188399 | GEO

Transcription profiling of mouse neurite outgrowth and regeneration (super series)

Project description:This SuperSeries is composed of the following subset Series:; GSE9738: Analysis of gene expression during neurite outgrowth and regeneration (430A and 420A 2.0 array); GSE9739: Analysis of gene expression during neurite outgrowth and regeneration (MG-U74A); GSE9740: Analysis of gene expression during neurite outgrowth and regeneration MG-U74B Experiment Overall Design: Refer to individual Series

2008-04-10 | E-GEOD-9744 | biostudies-arrayexpress

Schwann cell precursors represent a neural crest-like hub state with biased multipotency

Project description:Schwann cell precursors (SCPs) are nerve-associated progenitors that not only can generate myelinating and non-myelinating Schwann cells but also are multipotent like the neural crest cells from which they originate. SCPs are omnipresent along outgrowing peripheral nerves throughout the body of vertebrate embryos. By using single-cell transcriptomics to generate the atlas of the entire neural crest lineage, we show that early SCPs and late migratory crest have similar transcriptional profiles characterized by a multipotent “hub” state containing cells biased towards traditional neural crest fates. SCPs keep diverging from the neural crest after being primed towards terminal Schwann cell and other fates, with different subtypes residing in distinct anatomical locations. Functional experiments using CRISPR-Cas9 loss-of-function further show that knock-out of the hub gene Sox8 causes defects in neural crest-derived cells along peripheral nerves by facilitating differentiation of SCPs towards sympathoadrenal fates. Finally, specific tumour populations found in melanoma, neurofibroma and neuroblastoma map to different stages of SCP/Schwann cell development. Overall, SCPs resemble migrating neural crest cells that maintain multipotency and become transcriptionally primed toward distinct lineages.

2022-04-28 | GSE201257 | GEO

Phenotypic Screening Following Transcriptomic Deconvolution to Identify Transcription Factors Mediating Axon Growth Induced by a Kinase Inhibitor

Project description:After injury to the central nervous system (CNS), both neuron-intrinsic limitations on regenerative responses and inhibitory factors in the injured CNS environment restrict regenerative axon growth. Instances of successful axon regrowth offer opportunities to identify features that differentiate these situations from that of the normal adult CNS. One such opportunity is provided by the kinase inhibitor RO48, which dramatically enhances neurite outgrowth of neurons in vitro and substantially increased contralateral sprouting of corticospinal tract neurons when infused intraventricularly following unilateral pyramidotomy. The authors present here a transcriptomic deconvolution of RO48-associated axon growth, with the goal of identifying transcriptional regulators associated with axon growth in the CNS. Through the use of RNA sequencing (RNA-seq) and transcription factor binding site enrichment analysis, the authors identified a list of transcription factors putatively driving differential gene expression during RO48-induced neurite outgrowth of rat hippocampal neurons in vitro. The 82 transcription factor motifs identified in this way included some with known association to axon growth regulation, such as Jun, Klf4, Myc, Atf4, Stat3, and Nfatc2, and many with no known association to axon growth. A phenotypic loss-of-function screen was carried out to evaluate these transcription factors for their roles in neurite outgrowth; this screen identified several potential outgrowth regulators. Subsequent validation suggests that the Forkhead box (Fox) family transcription factor Foxp2 restricts neurite outgrowth, while FoxO subfamily members Foxo1 and Foxo3a promote neurite outgrowth. The authors’ combined transcriptomic-phenotypic screening strategy therefore allowed identification of novel transcriptional regulators of neurite outgrowth downstream of a multitarget kinase inhibitor.

2023-06-19 | GSE234696 | GEO

Herpes simplex virus type 1 modifies the protein composition of extracellular vesicles to promote neurite outgrowth and facilitate neuroinfection

Project description:Herpes simplex virus type 1 (HSV-1) is a highly contagious and prevalent human pathogen that causes many diseases, including encephalitis. During primary infection, HSV-1 infects epithelial cells and then neurites of peripheral neurons, establishing lifelong infection in the neuronal cell body. Neurites are highly dynamic structures that grow or retract in the presence of attractive or repulsive cues, respectively. Whether HSV-1 modulates these cues and thereby neurite outgrowth was not known. Here, we show that HSV-1 glycoprotein G (gG) reduces the inhibitory effect of epithelial cells on neurite outgrowth, facilitating viral infection of neurons through neurite ends. The mechanism of action involves the modification of the protein composition of extracellular vesicles (EV), including increased amount of galectin-1. We show that galectin-1 located in EV produced during HSV-1 infection of epithelial cells promotes neurite outgrowth, and this activity can be partially neutralized by antibodies. This study provides new insights into the neurotropism of HSV-1, identifying for the first time a viral protein that modifies the protein composition of EV to increase neurite outgrowth and neuronal infection.

2024-01-06 | PXD039569 | Pride

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