January 15-19, 2011
Town & Country Convention Center
San Diego, CA
Elane C Wright1 , Caixia Yang1,2 , Zhiqiang Du1,2 , Max F Rothschild1,2 , Jason W Ross1,2
During the peri-implantation stage of pregnancy, porcine conceptuses undergo rapid morphological rearrangement of their trophectoderm and initiate contact with the uterine endometrium to establish an initial interface necessary in the formation of a diffuse epitheliolchorial placenta. Numerous mRNA molecules are differentially expressed during this dynamic period of conceptus growth. To better understand mechanisms of post-transcriptional gene regulation during this process we conducted massively parallel deep sequencing of small RNA during conceptus elongation and the initial phase of uterine attachment. Day 12 spherical (8-10 mm; D12S), day 12 filamentous (greater than 100 mm in length; D12F) and day 14 filamentous (D14F) embryos were collected, extracted for small RNA and subjected to library construction using the Small RNA Expression Kit (Ambion). The libraries were then used for deep sequencing analysis using the SOLiD sequencing system (Applied Biosytems). Sequence reads were mapped to the human genome to determine the number of miRNA reads for each stage of development. Total reads for each library and the frequency of each miRNA within libraries were determined to identify differentially expressed miRNAs. During the transition from D12S to D12F, miR-301a, miR-1308, and miR-200a were increased in abundance, while miR-23b, miR-342, miR-30a and miR-10a were among those most abundantly expressed in D14F compared to earlier stages of development. These data demonstrated potential non-coding RNA markers of rapid trophoblastic elongation in the pig. This work was supported in part by the USDA National Institute of Food and Agriculture (2008-35205-05309).