Serial analysis of gene expression during elongation of the peri-implantation porcine trophectoderm (conceptus).
Conceptus loss during the preimplantation and early postimplantation period hinders the efficiency of swine reproduction. Significant conceptus loss occurs during trophectoderm elongation between gestational day 11 (D11) and day 12 (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 P-450(scc) (CYP11A1), aromatase (CYP19A), and steroidogenic acute regulatory protein (STAR)] and oxidative stress response pathways [microsomal glutathione S-transferase 1 (MGST1) and copper-zinc superoxide dismutase (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.
Blomberg
Le Ann
LA
Biotechnology and Germplasm Laboratory, United States Department of Agriculture Agricultural Research Service, Beltsville, Maryland 20705, USA. lblomberg@anri.barc.usda.gov
Long
Ezhou L
EL
Sonstegard
Tad S
TS
Van Tassell
Curtis P
CP
Dobrinsky
John R
JR
Zuelke
Kurt A
KA
D000818:Animals; D001141:Aromatase / Q000096:biosynthesis; D001755:Blastocyst; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis; D004475:Ectoderm / Q000378:metabolism*; D010064:Embryo Implantation; D020224:Expressed Sequence Tags; D015870:Gene Expression; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D005982:Glutathione Transferase / Q000096:biosynthesis; D008861:Microsomes / Q000201:enzymology; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress; D010641:Phenotype; D010750:Phosphoproteins / Q000096:biosynthesis; D012313:RNA / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000096:biosynthesis; D013552:Swine; D013997:Time Factors; D000098682:Steroidogenic Acute Regulatory Protein
D010750:Phosphoproteins; D000098682:Steroidogenic Acute Regulatory Protein; D012313:RNA; D001141:Aromatase; D002786:Cholesterol Side-Chain Cleavage Enzyme; D013482:Superoxide Dismutase; D005982:Glutathione Transferase