Project description:Recently, microRNAs (miRNAs) have emerged as new players in the fine tuning of embryo development and implantation in mammals via posttranscriptional gene regulation mechanisms. Applying custom made multispecies arrays we aimed to analyze expression profile of microRNAs in peri-implantation porcine conceptuses/trophoblasts to identify their potential role at the maternal-fetal interface during the critical period of maternal recognition of pregnancy and implantation. miRNA expression profiles were analyzed in samples collected from embryos or trophoblast on Days 10, 11, 12, 16 and 20 of pregnancy. Each group was represented by five to nine samples.
Project description:Affymetrix miRNA arrays were used to generate miRNA profiles of on the endometrium of Meishan and Yorkshire pigs on days 15 (implantation stage), 26 (post-implantation stage) and 50 (mid-gestation stage) of gestation. The study allowed for the determination of the miRNAs that were differentially expressed in porcine endometrium on different gestational day and breed. The majority of the miRNAs were differentially expressed during the implantation stage. This study will provide the information to better understanding of the role miRNAs in the porcine endometrium during pregnancy. the endometrium form different gestational day and breed were collected. In order to determine the miRNA expression profile of these tissue, RNA was extracted, labeled and hybridized on an Affymetrix miRNA array.
Project description:Environmental estrogens may affect epigenetic programming as early as the period of preimplantation development. Therefore, we analyzed the effects of continuous gestational estradiol-17β (E2) exposure on male and female embryos. A low dose, close to the no-observed effect level (NOEL - 10 µg E2/kg body weight(bw)/d), a high dose (1000 µg E2/kg bw/d) and carrier only, as control group, was fed to sows from insemination until sampling at day 10 of pregnancy, respectively. 36 samples (n = 5-7 per treatment group and sex) were analyzed by high throughput sequencing.In the high dose group, RNA-sequencing of single embryos revealed 982 differentially expressed genes (DEG) in the female but none in the male blastocysts. Moreover, 62 and 3 DEG were found in female and male embryos of the NOEL dose group, respectively. Thus, maternal E2 treatment during early pregnancy affected gene expression of the embryos at day 10, potentially constituting the basis for long-term adverse effects.
Project description: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
Project description:The study allowed for the determination of the mRNAs that were differentially expressed in porcine endometrium on different gestational day and breed. The majority of the mRNAs were differentially expressed during the implantation stage. This study will provide the information to better understanding of the role of these differently expressed mRNAs in the porcine endometrium during pregnancy. Affymetrix mRNA arrays were used to generate mRNA profiles of on the endometrium of Meishan and Yorkshire pigs on days 15 (implantation stage), 26 (post-implantation stage) and 50 (mid-gestation stage) of gestation. The endometrium from different gestational day and breed were collected. In order to determine the mRNA expression profile of these tissue, RNA was extracted, labeled and hybridized on an Affymetrix mRNA array.