Alterations in the transcriptome of porcine oocytes derived from prepubertal and cyclic females is associated with developmental potential.

The developmental competence of oocytes is progressively attained as females approach puberty. The poor quality of prepubertally derived oocytes suggests that essential processes during cytoplasmic maturation have not been completed. The objective of this experiment was to identify genes in oocytes that are associated with good (cyclic females) and poor (prepubertal females) developmental competence. Development to the blastocyst stage in vitro was significantly decreased in oocytes derived from prepubertal females compared with cyclic females (5.26 and 12.86%, respectively). Approximately 10% of the oocyte transcriptome was differentially expressed between in vitro-matured oocytes derived from cyclic and prepubertal females (P < 0.05); 58% of differentially expressed genes had increased transcript abundance in oocytes derived from cyclic females. Genes involved in the metabolism and regulation of biological processes had increased transcript abundance in oocytes derived from cyclic females, whereas genes involved in translation were increased in prepubertally derived oocytes. Quantitative PCR confirmed differential expression (P < 0.05) for 6 out of 11 selected genes [DPYD (dihydropyrimidine dehydrogenase), RDH11 (retinol dehydrogenase 11), SFRS4 (serine/arginine-rich splicing factor 4), SFRS7 (serine/arginine-rich splicing factor 7), TL4 (transcribed loci 4), and TOP2B (topoisomerase II ß)] that were differentially expressed with greater than a 2-fold change by microarray, although 3 of these genes, DPYD, TL4, and TOP2B, were in opposing directions by the 2 methods. In conclusion, expression of multiple genes involved in metabolism and translation was significantly altered in oocytes from prepubertal females compared with cyclic females, which was associated with reduced in vitro development to the blastocyst stage. These genes may represent important cellular mechanisms that regulate oocyte quality.

Developmental changes in insulin-like growth factor (IGF)-I and -II mRNA abundance in extra-embryonic membranes and small intestine of avian embryos.

OBJECTIVE: Numerous researchers have evaluated the insulin-like growth factors (IGF) influence on mammalian fetal development. Although IGF has been explored in the avian system, questions remain on the role of IGF in avian development. Therefore, the current study evaluated the mRNA abundance of IGF in the amnion and allantoic membranes and developing small intestine in the chicken, duck, and turkey during the incubation and post-hatch period. DESIGN: Broiler, duck, and turkey eggs were incubated with small intestinal, allantoic, and amniotic membranes collected in the final days of incubation and 1 week post-hatch. RNA was extracted using Trizol and qRT-PCR was utilized to compare differences during embryo development within and across species. RESULTS: The expression of the IGF mRNA varied between species in the final days of incubation in the amniotic and allantoic membranes. The turkey had higher (0.38-1.72 log) transcript abundance of IGF-I and IGF-II in the amnion and allantois compared to the chicken and duck. Evaluating the mRNA abundance within the chicken duodenum, jejunum, and ileum, the duodenum had the lowest expression of IGF-I and IGF-II (P<0.05) at day -4 of incubation compared to the jejunum and ileum. Focusing on differences in jejunal IGF expression among the three species, the turkey had the lowest IGF-I abundance at day -4 of incubation and highest IGF-I abundance at day of hatch (P<0.05). Transcript abundance of both IGF-II and IGF-R was highest in the turkey at day of hatch and day 1 post-hatch compared to the duck and chicken. The whole tissue versus the mucosal expression of the IGF mRNA abundance was evaluated during the post-hatch period. Duodenal, jejunal, and ileal segments had higher IGF-I transcript abundance (P<0.05) at day 1, day 3, and day of hatch, respectively. No differences were observed between segment and mucosa for IGF-II in the post-hatch period. The duodenal and jejunal mucosa IGF-R transcript abundance was greater (P<0.05) at day of hatch compared to the intestinal segment. The duck IGF mRNA in the jejunal mucosa was higher than the whole segment and decreased from day of hatch to day 3 post-hatch while the IGF mRNA abundance increased in the whole segment during the same time period. The turkey IGF-I transcript abundance decreased in both the segment and mucosa following hatch while the IGF-II mRNA expression increased by 1.5 logs from hatch to day 1 post-hatch. CONCLUSION: The transcript abundance of the IGF axis in the extra-embryonic membranes and gastrointestinal tissue of the developing chicken, duck, and turkey are influenced by embryonic age and species. A better understanding of the IGF axis in the small intestine during embryonic development may allow for increasing the optimal growth of both the gastrointestinal tract and the neonate.

Analysis of gene expression during the onset of muscle hypertrophy in callipyge lambs.

The callipyge mutation causes postnatal muscle hypertrophy in heterozygous lambs that inherit a paternal callipyge allele (+/CLPG). Our hypothesis was that the up-regulation of one or both of the affected paternally expressed genes (DLK1 or PEG11) initiates changes in biochemical and physiological pathways in skeletal muscle to induce hypertrophy. The goal of this study was to identify changes in gene expression during the onset of muscle hypertrophy to identify the pathways that are involved in the expression of the callipyge phenotype. Gene expression was analysed in longissimus dorsi total RNA from lambs at 10, 20, and 30 days of age using the Affymetrix Bovine Expression Array. An average of 40.6% of probe sets on the array was detected in sheep muscle. Data were normalized and analysed using a two-way anova for genotype and age effects with a false discovery rate of 0.10. From the anova, 13 genes were significant for the effect of genotype and 13 were significant for effect of age (P < 0.10). No significant age-by-genotype interactions were detected (P > 0.10). Of the 13 genes indicating an effect of genotype, quantitative PCR assays were developed for all of them and tested on a larger group of animals from 10 to 200 days of age. Nine genes had significantly elevated transcript levels in callipyge lambs. These genes included phosphofructokinase, a putative methyltransferase protein, a cAMP phosphodiesterase, and the transcription factor DNTTIP1.