The filipodia-like protrusions of adjacent somatic cells shape the developmental potential of mouse oocytes
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB56821 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR10682300.fastq.gz | Fastqsanger.gz | |||
| ERR10682301.fastq.gz | Fastqsanger.gz | |||
| ERR10682302.fastq.gz | Fastqsanger.gz | |||
| ERR10682303.fastq.gz | Fastqsanger.gz | |||
| ERR10682304.fastq.gz | Fastqsanger.gz |
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Project description:The oocyte must grow and mature before fertilization, thanks to a close dialogue with the somatic cells that surround it. Part of this communication is through filopodia-like protrusions, called transzonal projections (TZPs), sent by the somatic cells to the oocyte membrane. To investigate the contribution of TZPs to oocyte quality, we impaired their structure by generating a full knockout mouse of the TZP structural component myosin-X (MYO10). Using spinning disk and super-resolution microscopy combined with a machine-learning approach to phenotype oocyte morphology, we show that the lack of Myo10 decreases TZP density during oocyte growth. Reduction in TZPs does not prevent oocyte growth but impairs oocyte-matrix integrity. Importantly, we reveal by transcriptomic analysis that gene expression is altered in TZP-deprived oocytes and that oocyte maturation and subsequent early embryonic development are partially affected, effectively reducing mouse fertility. We propose that TZPs play a role in the structural integrity of the germline-somatic complex, which is essential for regulating gene expression in the oocyte and thus its developmental potential.
| S-EPMC10029974 | biostudies-literature
Project description:We used a fractionation scheme to isolate protrusions and cell bodies from control or APC knockdown fibroblasts, which were induced to extend protrusions in response to addition of lysophosphatidic acid (LPA). Four replicate protrusion and cell body samples for each cell type were analyzed by RNA-Seq. To identify RNAs enriched in protrusions, a protrusion/cell body ratio was calculated. We find that ca. 7% of detected RNAs were enriched in protrusions of control cells. Knockdown of APC reduced the protrusion enrichement of a subset of these RNAs, indicating that they are dependent on APC for their localization. Localization of the remaining RNAs was not significantly affected upon APC knockdown suggesting that their localization is mediated by an APC-independent mechanism.
2017-10-11 | GSE82229 | GEO
Project description:We generated fibroblast lines overexpressing beta-globin constructs under a doxycycline-inducible promoter. These constructs contain the beta-globin coding sequence followed by either a control 3'UTR (HBB) or the Pkp4 3'UTR. The Pkp4 3'UTR carries a protrusion-targeting element and is expected to compete with localization of endogenous protrusion-localized mRNAs. We used a fractionation scheme to isolate protrusions and cell bodies from HBB and Pkp4 UTR-expressing fibroblasts, which were induced to extend protrusions in response to addition of lysophosphatidic acid (LPA). 4 replicate protrusion and cell body samples for each cell type were analyzed by RNA-Seq. To identify RNAs enriched in protrusions, a protrusion/cell body ratio was calculated. We find that ca. 4.5% of detected RNAs were enriched in protrusions of control HBB cells. Overexpression of the Pkp4 UTR reduced the protrusion enrichement of a subset of these RNAs, while the degree of localization of the remaining RNAs was not significantly affected.
2017-10-11 | GSE82230 | GEO
Project description:Deciphering the complex biology of cell migration is key to understand human-related disorders. During angiogenesis, endothelial cells engage in coordinated migration events to form new blood vessels from parental counterparts. However, while subcellular localisation of mRNAs and localised translation are fundamental steps between gene transcription and protein activity, whether local regulation of gene expression controls the complex morphogenetic process of angiogenesis has never been addressed. Here, we set out to investigate the cytoplasmic distribution of mRNAs in migratory endothelial cells. We isolated RNA from endothelial cell protrusions from their cell bodies and used RNA-sequencing to identify asymmetrically distributed transcripts. These studies identified a set of transcripts enriched in endothelial cell protrusions over cell bodies, which included classically protrusion-enriched mRNAs and other intriguing transcripts encoding proteins implicated in cell migration.
2020-09-07 | GSE133055 | GEO
Project description:Like neurons, oligodendrocytes (OL) are cells with elaborate morphology that probably require asymmetrical spatial regulation of biological processes. Formation of membrane protrusions is critical for OL development and interaction with axons. We hypothesized that the enrichment of specific mRNAs in protrusions of oligodendrocyte precursor cells (OPC) is important for morphological differentiation, thus having an impact in myelination. To explore this hypothesis, we established a modified Boyden chamber system to physically separate soma from membrane protrusions of rat primary OPC cultured in vitro for 24h. We performed a whole transcriptome analysis (RNAseq) of primary rat OPC soma and membrane protrusion fractions and found a subcellular enrichment of mRNAs in these structures during initial protrusion formation. At the very initial stage of OPC protrusion extension, there is a significant subcellular enrichment of transcripts encoding proteins related to cellular component assembly and cytoskeleton organization, particularly of actin-related molecules. This suggests that the regulation of the cytoskeleton dynamics may be locally controlled in OPCs and probably relevant for their differentiation program.
2018-05-07 | GSE106862 | GEO
Project description:3' mRNA-seq of protrusions and cell bodies of nt or LARP6 siRNA transfected MDA-MB231 cells. These data show that Ribosomal protein (RP) -mRNAs localization to cell protrusions is mediated by LARP6.
2022-10-03 | E-MTAB-8472 | biostudies-arrayexpress
Project description:The mammalian oocyte has the unique feature of supporting fertilization and normal development while being able of reprogramming the nuclei of somatic cells towards pluripotency and occasionally even totipotency. Whilst oocyte quality is known to decay with somatic aging it is not a given that different biological functions decay concurrently. In this study we tested whether oocyte reprogramming ability decreases with somatic aging. We show that oocytes isolated from mice aged beyond the usual reproductive age (climacteric) yield ooplasts that retain reprogramming capacity after somatic nuclear transfer (SCNT) at levels similar to ooplasts of young donors. Despite differences in transcriptome between ooplasts of old and young mice gene expression profiles of the resultant SCNT blastocysts were very similar. Furthermore albeit results showing global down-regulation of apoptosis-related genes in climacteric ooplasts and the reported beneficial effect of inhibiting p53 on transcription factor-induced (iPS) pluripotency reprogramming capacity of young ooplasts was not affected by specifically blocking p53. Our observations strongly suggest that the outcome of oocyte-induced reprogramming is determined by the availability of intrinsic reprogramming factors tightly regulated throughout aging as well as intracellular and environmental checkpoints during pre-implantation development towards selection of the successfully reprogrammed embryos. While oocytes are not regenerated but rather last for life we further propose that these cells can still be a resource for somatic reprogramming when they cease to be considered safe for sexual reproduction. Total mRNA was isolated for samples from young and climacteric mice (pools of 20 ooplasts and 20 SCNT blastocysts in duplicate) using a RNeasy Mini Kit (Qiagen Valencia CA USA) as described by the manufacturer (no DNase treatment). RNA concentrations as well as purity and integrity check were determined with Agilent Bioanalyzer 2100 and RNA Pico 6000 Lab-Chip Kit (Agilent Technologies Palo Alto CA USA). RiboAmp HS Plus Amplification Kit (MDS Analytical Technologies Ismaning Germany) was used to amplify the total mRNA with two rounds of amplification according to the manufacturerâ??s instructions. Amplified RNA was eluted with 15 µL RE Buffer included in the RiboAmp HS Plus Amplification Kit. RNA concentrations were determined with Agilent Bioanalyzer 2100 and RNA Pico 6000 Lab-Chip Kit. Turbo Labeling CY3 Kit (MDS Analytical Technologies) was used to label 3 µg of amplified RNA with Cy3. Concentration and frequency of incorporation were measured with NanoPhotometer (Implen). Microarray wash and detection of the labeled RNA on GeneChips were carried out according to the manufacturerâ??s instructions (Agilent Technologies). Gene expression profiling was performed using Agilent Whole Mouse Genome Oligo Microarrays (4 x 44k each array containing 41174 features). Array image acquisition and feature extraction was performed using Agilent G2505B Microarray Scanner and Feature Extraction software v.9.5 (Agilent Technologies). 6 samples were analyzed. YoungOoplasts: young ooplasts 1 biological rep ClimactericOoplasts: climacteric ooplasts 1 biological rep YoungBlastocysts: young blastocysts 2 biological rep ClimactericBlastocysts: climacteric blastocysts 2 biological rep
2011-08-15 | E-GEOD-23654 | biostudies-arrayexpress
Project description:The Scar/WAVE regulatory complex (WRC) is the main catalyst of pseudopod and lamellipodium formation during cell migration. Its actin nucleation activity has been attributed to its ability to activate the Arp2/3 complex through the VCA domain of Scar/WAVE. Here we show in both B16-F1 mouse melanoma cells and Dictyostelium discoideum cells that the WRC with its VCA domain deleted can still induce the formation of morphologically normal actin protrusions. Its function is lost only after further deletion of Scar/WAVE’s (in Dictyostelium cells) or both Abi and WAVE’s (in B16-F1 cells) proline-rich domains. Moreover, mass spectrometry analysis identified BAIAP2 and Vasp (both promotors of actin polymerization) as specific interactors of WRC polyproline domains. Thus we conclude that proline-rich domains play a central role in actin nucleation by concentrating other effectors needed to form actin protrusions. Together, these findings suggest a new mechanism for WRC action independent of the VCA-Arp2/3 interaction.
2024-08-30 | PXD037123 | Pride
Project description:3' mRNA-seq of protrusions and cell bodies of a panel of normal and malignant cell lines. These data show that Ribosomal protein (RP) -mRNAs localization to cell protrusions is a conserved phenomenon.
2020-10-30 | E-MTAB-8470 | biostudies-arrayexpress
Project description:Cell-cell communication is vital to multicellular organisms, and distinct types of cellular protrusions play critical roles during development, cell signaling, and the spreading of pathogens and cancer. The differences in the structure and protein composition of these different types of protrusions that could explain their specific functions have not been elucidated due to the lack of a method for their specific isolation and analysis. In this paper, we describe, for the first time, a method to specifically isolate distinct protrusion subtypes, based on their morphological structures or fluorescent markers, using laser capture microdissection (LCM). Combined with a unique fixation and protein extraction protocol, we have pushed the limits of microproteomics and demonstrate that proteins from LCM isolated protrusions can successfully and reproducibly be identified by mass spectrometry using ultra-high field Orbitrap technologies. Our method confirms that different types of protrusions have distinct proteomes and it promises to advance the characterization and understanding of these unique structures.
2018-07-05 | MSV000082576 | MassIVE