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Peri-implantation gestational day 11 porcine embryos

ABSTRACT: Asynchrony of trophectoderm elongation, gestational days 11-12, is evident in porcine concepti and, rapid progression through this phase has been associated with conceptus competency. The goal of the current study was to determine the extent of transcriptomic responses of concepti to developmental delay and their physiological implications. Concepti with the same morphology, ovoid and 7-8 mm, were isolated and designated as normal or developmentally delayed if collected from a homogenous ovoid conceptus population or a heterogeneous conceptus population (ovoid to filamentous), respectively. Total RNA prepared from 4 distinct normal and 4 distinct developmentally delayed concepti, was analyzed using an Agilent high-density custom porcine microarray. Two hundred-nine transcripts were found differentially expressed between normal and developmentally delayed concepti. Functional analysis of these genes indicated that a significant number of the genes regulate signal transduction/transcription, organismal development, metabolism, and cell adhesion and can be modulated by transforming growth factor β1. Ten genes were selected for real-time PCR validation of differential expression based on a known role in steroid synthesis, endometrium receptivity, and modulation of trophoblast differentiation/growth or interaction with transforming growth factor β1. As in the microarray, all except one, achaete-scute complex homolog 2, were preferentially up-regulated in delayed concepti. Overall, the findings suggested that developmentally delayed concepti are distinct from their morphologically similar counterparts and more developmentally advanced concepti. Furthermore, it also highlighted ways by which the conceptus’ microenvironment might be affected and pinpointed developmental factors that can be interrogated further to determine if, and how, they affect embryo competency/elongation.

ORGANISM(S): Sus scrofa

PROVIDER: GSE18472 | GEO | 2009/12/10

SECONDARY ACCESSION(S): PRJNA118203

REPOSITORIES: GEO

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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.

Project description:Venkatraman2012 - Interplay between PLS and TSP1 in TGF-β1 activation The interplay between PLS (Plasmin) and TSP1 (Thrombospondin-1) in TGF-β1 (Transforming growth factor-β1)is shown using mathematical modelling and in vitro experimentents. This model is described in the article: Plasmin triggers a switch-like decrease in thrombospondin-dependent activation of TGF-β1. Venkatraman L, Chia SM, Narmada BC, White JK, Bhowmick SS, Forbes Dewey C Jr, So PT, Tucker-Kellogg L, Yu H. Biophys J. 2012 Sep 5;103(5):1060-8. Abstract: Transforming growth factor-β1 (TGF-β1) is a potent regulator of extracellular matrix production, wound healing, differentiation, and immune response, and is implicated in the progression of fibrotic diseases and cancer. Extracellular activation of TGF-β1 from its latent form provides spatiotemporal control over TGF-β1 signaling, but the current understanding of TGF-β1 activation does not emphasize cross talk between activators. Plasmin (PLS) and thrombospondin-1 (TSP1) have been studied individually as activators of TGF-β1, and in this work we used a systems-level approach with mathematical modeling and in vitro experiments to study the interplay between PLS and TSP1 in TGF-β1 activation. Simulations and steady-state analysis predicted a switch-like bistable transition between two levels of active TGF-β1, with an inverse correlation between PLS and TSP1. In particular, the model predicted that increasing PLS breaks a TSP1-TGF-β1 positive feedback loop and causes an unexpected net decrease in TGF-β1 activation. To test these predictions in vitro, we treated rat hepatocytes and hepatic stellate cells with PLS, which caused proteolytic cleavage of TSP1 and decreased activation of TGF-β1. The TGF-β1 activation levels showed a cooperative dose response, and a test of hysteresis in the cocultured cells validated that TGF-β1 activation is bistable. We conclude that switch-like behavior arises from natural competition between two distinct modes of TGF-β1 activation: a TSP1-mediated mode of high activation and a PLS-mediated mode of low activation. This switch suggests an explanation for the unexpected effects of the plasminogen activation system on TGF-β1 in fibrotic diseases in vivo, as well as novel prognostic and therapeutic approaches for diseases with TGF-β dysregulation. This model is hosted on BioModels Database and identified by: MODEL1303130000 . To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models . To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

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Peri-implantation gestational day 11 porcine embryos

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