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Pig peri-implantation embryo development transcriptional profiling

ABSTRACT: A novel porcine cDNA microarray, containing PCR products selected for embryonic expression, was used for transcriptional profiling during conceptus development. Total RNA from quadruplicate samples of small spherical (1-5mm), large spherical (6-11mm), tubular (13-45mm), and filamentous (>100mm) was extracted, amplified, and converted to 32P labeled target and hybridized to membranes. Porcine peri-implantation embryo development order is from small spherical to large spherical, then to Tubular, then to filamentous. All this morphological transmission is within two days (day 10 to day 12). In this study, we investigated the gene expression changes by a specifically selected porcine embryo gene array. A number of genes were found to be differentially expressed during those developmental stages and confirmed by quantitative PCR. Keywords = Gene expression Keywords = porcine conceptus Keywords = cDNA microarray Keywords = peri-implantation Keywords = transcriptional profiling Keywords: time-course

ORGANISM(S): Sus scrofa

PROVIDER: GSE1372 | GEO | 2004/05/13

SECONDARY ACCESSION(S): PRJNA90135

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.

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Pig peri-implantation embryo development transcriptional profiling

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