ARS | Publication request: SERIAL ANALYSIS OF GENE EXPRESSION (SAGE) DURING ELONGATION OF PERIIMPLANTATION PORCINE EMBRYOS

Submitted to: Experimental Biology
Publication Type: Abstract
Publication Acceptance Date: March 15, 2003
Publication Date: April 11, 2003
Citation: Blomberg, L., Long, E.L., Sonstegard, T.S., Van Tassell, C.P., Dobrinsky, J.R., Zuelke, K.A. 2003. Serial analysis of gene expression (sage) during elongation of periimplantation porcine embryos. Experimental Biology.

Technical Abstract: In the pig, ~ 30% of embryos are lost prior to gestational day 20; the major loss occurs between day 11 (D11) and day 12 (D12) when embryos undergo rapid elongation prior to implantation. Elucidating the array of genes expressed in D11 and D12 embryos would provide insight regarding essential temporally regulated genes, which could serve as potential markers of development efficacy. To determine qualitative and quantitative gene expression of D11 and D12 embryos, SAGE was done. Total RNA from in vivo derived D11 and D12 embryos was used to construct individual SAGE tag libraries. Analysis of the libraries yielded a total of 42,389 tags (D11) and 42,392 tags (D12) representing 14,464 and 13,098 putative genes, respectively. Comparative statistical analysis of the SAGE tag frequencies indicated that at p<0.05 significance and p<0.001 significance, 434 tags and 87 tags, respectively, were differentially expressed between D11 and D12. The SAGE tags, annotated following blasts on NCBI and TIGR species indices databases, were clustered into functional groups. Real-time PCR confirmed differential expression of several transcripts between D11 and D12. Despite the paucity of porcine sequence data, SAGE proved effective in identifying genes potentially crucial to embryo development and should aid in providing a more comprehensive picture of the porcine transcriptome. Supported by USDA ARS CRIS Project No. 1265-31000-082.