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Physiol. Genomics (November 9, 2004). doi:10.1152/physiolgenomics.00157.2004
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Submitted on July 22, 2004
Accepted on November 5, 2004

Serial Analysis of Gene Expression (SAGE) During Elongation of the Peri-Implantation Porcine Trophectoderm (Conceptus)

Le Ann Blomberg1*, Ezhou L Long2, Tad S Sonstegard3, Curtis P Van Tassell3, John R Dobrinsky1, and Kurt A Zuelke1

1 Animal and Natural Resources Institute, Biotechnology & Germplasm Laboratory, USDA Agricultural Research Service, Beltsville, MD, USA
2 Animal and Natural Resources Institute, Biotechnology & Germplasm Laboratory, USDA Agricultural Research Service, Beltsville, MD, USA; Mammary Biology &Tumorigenesis Laboratory, National Cancer Institute, Bethesda, MD, USA
3 ANRI, Bovine Functional Genomics Laboratory, USDA, Agricultural Research Service, Beltsville, MD, USA

* To whom correspondence should be addressed. E-mail: lblomberg{at}anri.barc.usda.gov.

Conceptus loss during the pre-implantation and early post-implantation period hinders the efficiency of swine reproduction. Significant conceptus loss occurs during trophectoderm elongation between gestational day (D) 11 and D12. Elongation of the porcine conceptus is a key stage of development during which maternal recognition of pregnancy, initial placental development, and preparation for implantation occurs. The objective of this study was to establish comparative transcriptome profiles of D11 ovoid and D12 filamentous conceptuses and thereby identify temporally regulated genes essential for developmental progression during conceptus elongation. Serial analysis of gene expression (SAGE) libraries were constructed from in vivo derived ovoid and filamentous swine conceptuses to yield a total of 42,389 tags (ovoid) and 42,391 tags (filamentous) representing 14,464 and 13,098 putative unique transcripts, respectively. Statistical analysis of tag frequencies revealed the differential expression of 431 tags between libraries (p<0.05). Nucleotide sequence alignment searches on public databases provided SAGE tag annotation and gene ontology assignments. Comparisons between the SAGE profiles of ovoid and filamentous conceptuses revealed increased expression of key genes in the steroidogenesis (cytochrome p450scc (CYP11A1), aromatase (CYP19A), and steroidogenic acute regulatory protein (STAR)) and oxidative stress response pathways (microsomal glutathione S-transferase 1 (MGST1) and copper-zinc SOD (SOD1)). Differential expression of these genes in the steroidogenic and oxidative stress response pathways was confirmed by real time PCR. These results validate the utility of SAGE in the pig and establish an initial model linking gene expression profiles at the pathway level with phenotypic progression from ovoid to filamentous stages of conceptus development.




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