Transcriptional profiling by deep sequencing indentifies [sic] differences in mRNA transcript abundance in in vivo derived vs. in vitro cultured porcine blastocyst stage embryos

MOspace/Manakin Repository

Breadcrumbs Navigation

Transcriptional profiling by deep sequencing indentifies [sic] differences in mRNA transcript abundance in in vivo derived vs. in vitro cultured porcine blastocyst stage embryos

Please use this identifier to cite or link to this item: http://hdl.handle.net/10355/8098

[-] show simple item record

dc.contributor.advisor Prather, Randall S. en_US
dc.contributor.author Bauer, Bethany Kay en_US
dc.date.accessioned 2010-08-03T19:18:42Z
dc.date.available 2010-08-03T19:18:42Z
dc.date.issued 2010 en_US
dc.date.submitted 2010 Spring en_US
dc.identifier.other BauerB-050410-T3267 en_US
dc.identifier.uri http://hdl.handle.net/10355/8098
dc.description Title from PDF of title page (University of Missouri--Columbia, viewed on June 3, 2010). en_US
dc.description The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. en_US
dc.description Thesis advisor: Dr. Randall Prather. en_US
dc.description Includes bibliographical references. en_US
dc.description M.S. University of Missouri--Columbia 2010. en_US
dc.description Dissertations, Academic -- University of Missouri--Columbia -- Animal sciences. en_US
dc.description.abstract In vitro embryo culture systems promote development at rates lower than in vivo. The goal of this project was to discover transcripts that may be responsible for a decrease of embryo competency in blastocyst stage embryos cultured in vitro. Gilts were artificially inseminated and on Day 2 one oviduct and the tip of a uterine horn were flushed and the recovered embryos were cultured in PZM3 for four days. On Day 6 the gilts were euthanized and the contra-lateral horn was flushed to obtain in vivo derived embryos. Total RNA was extracted from 3 pools of 10 blastocysts from each treatment. First and second strand cDNA was synthesized and then sequenced using Illumina sequencing. The reads were aligned to a custom-built database designed to represent the known porcine "transcriptome". A total of 1,170 database members were different between the two groups (P[less than]0.05), and 588 of those were at least 2-fold different. Eleven transcripts were subjected to real-time PCR and each validated the sequencing. There was an overall decrease in both inner cell mass and trophectodermal cell numbers in embryos cultured in vitro, however, no difference in ICM:TE ratio was found. Interestingly, the transcript (SLC7A1) was higher in the in vitro cultured group. This difference disappeared after addition of arginine (0.36 mM) to the 4 day culture. In conclusion, Illumina sequencing and alignment to a custom "transcriptome" successfully identified a large number of genes that yield clues to the derivation of culture media. en_US
dc.format.extent xi, 88 pages en_US
dc.language.iso en_US en_US
dc.publisher University of Missouri--Columbia en_US
dc.subject.lcsh Swine -- Embryos en_US
dc.subject.lcsh Swine -- Breeding en_US
dc.subject.lcsh Fertilization in vitro en_US
dc.subject.lcsh Ovum implantation en_US
dc.subject.lcsh Genetic transcription en_US
dc.subject.lcsh Nucleotide sequence en_US
dc.title Transcriptional profiling by deep sequencing indentifies [sic] differences in mRNA transcript abundance in in vivo derived vs. in vitro cultured porcine blastocyst stage embryos en_US
dc.type Thesis en_US
thesis.degree.discipline Animal sciences en_US
thesis.degree.grantor University of Missouri--Columbia en_US
thesis.degree.name M.S. en_US
thesis.degree.level Masters en_US
dc.identifier.merlin b77767317
dc.identifier.oclc 646494863 en_US
dc.relation.ispartofcommunity University of Missouri-Columbia. Graduate School. Theses and Dissertations. Theses. 2010 Theses
dc.relation.ispartofcollection 2010 Freely available theses (MU)


This item appears in the following Collection(s)

[-] show simple item record