Early mammalian development before the oocyte-to-embryo transition is under 'maternal control' from factors deposited in the cytoplasm during oocyte growth, synthesized independent of de novo transcription. Maternal mRNAs encode proteins necessary for early embryo development. Two elements in the mRNA 3’untranslated region (UTR), the cytoplasmic polyadenylation element (CPE) and the hexanucleotide (AAUAAA) are involved in the control of translation of specific mRNAs during meiotic maturation. Despite advances in understanding the translational regulation during meiotic maturation, regulation at the 1-cell stage has not been explained. More studies are required to explain this complex mechanism of temporal mRNA recruitment after fertilization. Maternal mRNAs translated at different stages were examined to understand how specific maternal mRNAs are synthesized and stored, what are these maternal mRNAs, which maternal mRNAs are translated, and how these maternal mRNAs are temporally regulated. Polysomal mRNAs from eggs and 1-cell embryos were analyzed by microarray analysis and this indicated that temporally significant biological activities were encoded by mRNAs recruited at different stages of development. The mRNAs recruited in eggs were involved in homeostasis and transport mechanisms and those recruited in zygotes were involved in biosynthesis and metabolic activities. These data indicated that there is a temporal regulation of maternal mRNAs to meet the different biological requirements of the embryos. After the identification of temporally translated mRNAs, experiments were performed to understand the mechanism underlying temporal translation. The prevalence of the CPE differed between the two mRNA populations translated i.e., egg and 1-cell stage polysomal mRNAs. CPEs were present in ~53% of transcripts at the 1-cell stage compared to ~86% at the MII stage. This indicated that novel motifs other than CPEs regulate translation of mRNAs at the 1-cell stage. Truncation and deletion experiments were conducted using chimeric mRNAs based on one mRNA that was enriched in the 1- cell polysomes (Bag4). These experiments led to the identification of two regulatory regions that control translation at the 1-cell stage, an 80 nt region and a 43 nt region with different regulatory motifs. The 80 nt region is involved in activation of translation and the 43 nt region has an inhibitory effect on translation at the MII and early 1-cell stage. These results provide a detailed picture of how specific maternal mRNAs are prevented from undergoing translation at the MII stage and how the effect of inhibition is eliminated by the late 1-cell stage. / Biochemistry
| Identifer | oai:union.ndltd.org:TEMPLE/oai:scholarshare.temple.edu:20.500.12613/2180 |
| Date | January 2010 |
| Creators | Potireddy, Santhi |
| Contributors | Latham, Keith, Sapienza, Carmen, Soprano, Dianne R., Shore, Scott K., Engel, Nora |
| Publisher | Temple University. Libraries |
| Source Sets | Temple University |
| Language | English |
| Detected Language | English |
| Type | Thesis/Dissertation, Text |
| Format | 134 pages |
| Rights | IN COPYRIGHT- This Rights Statement can be used for an Item that is in copyright. Using this statement implies that the organization making this Item available has determined that the Item is in copyright and either is the rights-holder, has obtained permission from the rights-holder(s) to make their Work(s) available, or makes the Item available under an exception or limitation to copyright (including Fair Use) that entitles it to make the Item available., http://rightsstatements.org/vocab/InC/1.0/ |
| Relation | http://dx.doi.org/10.34944/dspace/2162, Theses and Dissertations |
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