Understanding Bovine Embryo Elongation: A Transcriptomic Study of Trophoblastic Vesicles
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PROVIDER: PRJNA612017 | ENA |
REPOSITORIES: ENA
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Project description:Background: During the process of elongation, the embryo increases in size within the uterus while the extra-embryonic tissues (EET) develop and differentiate in preparation for implantation. As it grows, the ovoid embryo changes shape into first a tubular and then a filamentous form. This process is directed by numerous genes and pathways, the expression of which may be altered in the case of developmental irregularities such as when the conceptus is shorter than expected or when the embryo develops after splitting. In bovines, efforts to understand the molecular basis of elongation have employed trophoblastic vesicles (TVs)—short tubular EET pieces lacking an embryo—which also elongate in vivo. To date, however, we lack molecular analyses of TVs at the ovoid or filamentous stages that might shed light on the expression changes involved. Methods: Following in vivo development, we collected bovine conceptuses from the ovoid (D12) to filamentous stages (D18), sectioned them into small pieces with or without their embryonic disc (ED), and then transferred them to a receptive bovine uterus to assess their elongation abilities. We also grew spherical blastocysts in vitro up to D8, and subjected them to the same treatment. We then assessed differences in gene expression between the different samples and fully elongating controls at different stages of elongation using a bovine array (10K) and an extended qPCR array comprising 224 genes across 24 pathways. Results: In vivo, TVs elongated more or less depending on the stage at which they had been created and the time spent in utero. Their daily elongation rates differed from control EET, with the rates of TVs sometimes resembling those of earlier-stage EET. Overall, the molecular signatures of TVs followed a similar developmental trajectory as intact EET from D12–D18. However, within each stage, TVs and intact EET displayed distinct expression dynamics, some of which were shared with other short epithelial models. Conclusions: Differences between TVs and EET likely result from multiple factors, including a reduction in the length and signaling capabilities of TVs, delayed elongation from inadequate uterine signals, and modified crosstalk between the conceptus and the uterus. These findings confirm that close coordination between uterine, embryonic, and extraembryonic tissues is required to orchestrate proper elongation and, based on the partial differentiation observed, raise questions about the presence/absence of certain developmental cues or even their asynchronies.
2024-01-11 | GSE146768 | GEO
Project description:Background: During the process of elongation, the embryo increases in size within the uterus, while the extra-embryonic tissues (EETs) develop and differentiate in preparation for implantation. As it grows, the ovoid embryo transforms into a tubular form first and then a filamentous form. This process is directed by numerous genes and pathways, the expression of which may be altered in the case of developmental irregularities such as when the conceptus is shorter than expected or when the embryo develops after splitting. In bovines, efforts to understand the molecular basis of elongation have employed trophoblastic vesicles (TVs)-short tubular EET pieces that lack an embryo-which also elongate in vivo. To date, however, we lack molecular analyses of TVs at the ovoid or filamentous stages that might shed light on the expression changes involved. Methods: Following in vivo development, we collected bovine conceptuses from the ovoid (D12) to filamentous stages (D18), sectioned them into small pieces with or without their embryonic disc (ED), and then, transferred them to a receptive bovine uterus to assess their elongation abilities. We also grew spherical blastocysts in vitro up to D8 and subjected them to the same treatment. Then, we assessed the differences in gene expression between different samples and fully elongating controls at different stages of elongation using a bovine array (10 K) and an extended qPCR array comprising 224 genes across 24 pathways. Results: In vivo, TVs elongated more or less depending on the stage at which they had been created and the time spent in utero. Their daily elongation rates differed from control EET, with the rates of TVs sometimes resembling those of earlier-stage EET. Overall, the molecular signatures of TVs followed a similar developmental trajectory as intact EET from D12-D18. However, within each stage, TVs and intact EET displayed distinct expression dynamics, some of which were shared with other short epithelial models. Conclusion: Differences between TVs and EET likely result from multiple factors, including a reduction in the length and signaling capabilities of TVs, delayed elongation from inadequate uterine signals, and modified crosstalk between the conceptus and the uterus. These findings confirm that close coordination between uterine, embryonic, and extra-embryonic tissues is required to orchestrate proper elongation and, based on the partial differentiation observed, raise questions about the presence/absence of certain developmental cues or even their asynchronies.
| S-EPMC10859461 | biostudies-literature
Project description:The objective of the present investigation was to utilize the GeneChip® Porcine Genome Array from Affymetrix possessing 20, 201 unique probe sets to identify differentially expressed genes during rapid trophoblastic elongation and attachment to the uterine surface in the pig. Identification and characterization of conceptus gene expression patterns during rapid trophoblastic elongation and attachment in the pig will provide a better understanding of the events required for successful implantation and embryonic survival.
2008-09-10 | GSE12705 | GEO
Project description:The objective of the present investigation was to utilize the GeneChip® Porcine Genome Array from Affymetrix possessing 20, 201 unique probe sets to identify differentially expressed genes during rapid trophoblastic elongation and attachment to the uterine surface in the pig. Identification and characterization of conceptus gene expression patterns during rapid trophoblastic elongation and attachment in the pig will provide a better understanding of the events required for successful implantation and embryonic survival. Experiment Overall Design: Four chips were used for each morphological stage of development (spherical, tubular, Day 12 filamentous and Day 14 filamentous). RNA utilized for each chip represented a unique pool of conceptus total RNA for the respective morphological stage of development. Prior to target labeling, RNA was further purified (RNeasy MinElute Cleanup, Qiagen, Valencia, CA). Target labeling, GeneChip® hybridization, scanning and quantification were conducted by The University of Tulsa Microarray Core Facility. Affymetrix GeneChip Operating Software (GCOS version 1.1.1, Affymetrix, Santa Clara, CA) was used to quantitate each GeneChip®. The summary intensities for each probe were loaded into DNA-Chip Analyzer (dChip), version 1.3 for normalization, standardization, and analysis.
2008-10-25 | E-GEOD-12705 | biostudies-arrayexpress
Project description:The human placenta is covered by a single multinucleated fetal cell, the syncytiotrophoblast, which is bathed in maternal blood. During all pregnancies, membrane enclosed extracellular vesicles derived from the syncytiotrophoblast are extruded into the maternal blood.The large size of these extracellular vesicles (diameter larger than 10µm) is referred to as trophoblastic debris in this study. We have shown in the past that endothleial cells are involved in clearence of this trophoblastic debris and induction of immune tolerence by trophoblastic debris.This study aimed to characterise the transcriptional changes that occur in human vascular endothelial cells following exposure to trophoblastic debris from normal first trimester placentae. Microarrays were used to probe transcriptomic changes 2 and 21 hours after exposure of endothelial cells (Human microvascular endothelial cell line,HMEC-1) to trophoblastic debris from normal first trimester placentae Trophoblastic debris were isolated by low speed centrifugation from three individual first trimester human placentae (three biological replicates). The protein content in trophoblastic debris was measured by BCA assay. HMEC-1 was co-cultured with trophoblastic debris (60ug/ml total debris protein contents) for either 2 or 21 hours before RNA extraction. Untreated HMEC-1 at 2 and 21 hours were used as controls.In total, 12 samples were analyzed.
2016-08-21 | E-GEOD-69996 | biostudies-arrayexpress
Project description:The human placenta is covered by a single multinucleated fetal cell, the syncytiotrophoblast, which is bathed in maternal blood. During all pregnancies, membrane enclosed extracellular vesicles derived from the syncytiotrophoblast are extruded into the maternal blood.The large size of these extracellular vesicles (diameter larger than 10µm) is referred to as trophoblastic debris in this study. We have shown in the past that endothleial cells are involved in clearence of this trophoblastic debris and induction of immune tolerence by trophoblastic debris.This study aimed to characterise the transcriptional changes that occur in human vascular endothelial cells following exposure to trophoblastic debris from normal first trimester placentae. Microarrays were used to probe transcriptomic changes 2 and 21 hours after exposure of endothelial cells (Human microvascular endothelial cell line,HMEC-1) to trophoblastic debris from normal first trimester placentae
2016-08-21 | GSE69996 | GEO
Project description:Effect of DKK1 on embryo elongation
| PRJNA522975 | ENA
Project description:Extracellular vesicles (EVs) have emerged as key messengers in intercellular communications. However, the precise mechanisms of how recipient cells read EV messages are not clear. In this study, we investigated the roles of EV protein cargo, EV parent cell type, and recipient cell type in EV function using a human embryo implantation model. The JAr cell line served as an analog of human trophoblast cells, and EVs isolated from these cells were introduced into receptive endometrial epithelial cell analog RL95-2 cells to simulate EV-mediated embryo-maternal communication. Downregulation of Zinc-finger protein 81 (ZNF81) gene expression in RL95-2 cells in response to trophoblast EVs were used as a readout of JAr EV functionality. EVs from the non-trophoblastic cell line (HEK-293 cells) were used as a control to assess the specificity of trophoblast EV functionality in endometrial epithelial cells. Our results demonstrated that ZNF81 gene downregulation is a specific response of RL95-2 cells to JAr EVs, as HEK-293 cell EVs did not induce any gene expression changes. Furthermore, we conducted an analysis of the proteomic cargo profiles of JAr and HEK-293 cell EVs. Interestingly, JAr EVs exhibited a unique proteomic cargo profile compared to that of HEK EVs. Some of the proteins uniquely identified in JAr EVs are also known to play roles in embryo implantation. Moreover, JAr EVs were specifically enriched with important transcription factors that regulate ZNF81 gene expression. These findings contribute to our understanding of the specificity of EV functionality and cellular response in the context of embryo implantation.
2024-07-03 | PXD050024 | Pride
Project description:This project aims at identifying missing proteins in naive pluripotent and trophoblastic stem cells in the framework of the human proteome project (HPP). To date, as much as 1343 missing proteins remain to be credibly identified essentially, but not entirely, by mass spectrometry. The present results also constitute a first step towards the identification of potential biological markers for assessing stem cells and early embryo development.
2023-03-11 | PXD035768 | Pride
Project description:Objective To investigate the influence that oviductal extracellular vesicles from patients with endometriosis exert on early embryo development. Design In vitro experimental study Setting University-affiliated hospital. Patient(s) Women with and without endometriosis who underwent hysterectomy (n = 15 in total). Intervention(s) None. Main Outcome Measure(s) Oviductal extracellular vesicles from patients with endometriosis (oEV-EMT) or without endometriosis (oEV-ctrl) were isolated and co-cultured with two-cell murine embryos for 75 hours. Blastocyst rates were recorded. RNA sequencing was used to identified the differentially expressed genes in blastocysts cultured either with oEV-EMT or with oEV-ctrl. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed to identify potential biological processes in embryos that oEV-EMT affect. The functions of oEV on early embryo development were determined by reactive oxygen species (ROS) levels, mitochondrial membrane potentials, total cell numbers and apoptotic cell proportions. Result(s) Extracellular vesicles were successfully isolated from human Fallopian tubal fluid and their characterizations were described. The blastocyst rates were significantly decreased in the oEV-EMT group. RNA sequencing revealed that oxidative phosphorylation was down-regulated in blastocysts cultured with oEV-EMT. Analysis of oxidative stress and apoptosis at the blastocysts stage showed that embryos cultured with oEV-EMT had increased ROS level, decreased mitochondrial membrane potentials, and increased apoptotic index. Total cell numbers were not influenced. Conclusion(s) Oviductal extracellular vesicles from patients with endometriosis exert a negative influence on early embryo development.
2023-06-02 | GSE225323 | GEO