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Influence of Estradiol on In vitro Maturation of Porcine Oocytes

dc.contributor.advisorCarl Lessarden_US
dc.contributor.committeeMemberJohn Hardingen_US
dc.contributor.committeeMemberAli Honaramoozen_US
dc.contributor.committeeMemberGregg Adamsen_US
dc.contributor.committeeMemberMichael Dycken_US
dc.creatorLeavins, Nikki Leeen_US
dc.date.accessioned2011-09-14T16:23:38Zen_US
dc.date.accessioned2013-01-04T04:58:23Z
dc.date.available2012-10-06T08:00:00Zen_US
dc.date.available2013-01-04T04:58:23Z
dc.date.created2011-01en_US
dc.date.issued2011-01-01en_US
dc.date.submittedJanuary 2011en_US
dc.description.abstractIn vitro production of embryos allows efficient management of herd genetics, reduction of disease impact, and if used in combination with other reproductive technologies it could aid in preserving the threatened genetic diversity of swine. In vitro maturation (IVM) is identified as a deficient step in porcine in vitro production (IVP) of embryo systems, which decreases the overall success of IVP. There are problems encountered in each step of IVP; chromosomal abnormalities and decreased cell numbers in blastocysts during in vitro culturing (IVC), and low monospermic fertilization rates during in vitro fertilization (IVF) may be a result of insufficient IVM. As an addition to maturation media, porcine follicular fluid (pFF) can affect IVM. Estrogen can be found in high concentrations in pFF; possibly contributing to the effects seen when pFF is added to IVM. The objective of this thesis was to investigate the effects of estrogen supplementation during IVM on IVP of porcine embryos. The first objective was to evaluate the in vitro maturation rates of porcine oocytes in two maturation media: protein-free and 10% pFF supplemented. Nuclear maturation of oocytes was evaluated using Lamin/Dapi staining of oocytes matured in protein-free and 10% pFF maturation media to ensure the efficiency of the protein-free media. Protein-free and 10% pFF media mature oocytes at similar rates (91% and 89% respectively). The transcripts within the oocyte can be altered based on the in vitro maturation environment, so the second objective was to observe the expression of four chosen maternal effect genes: Basonuclin-1 (BNC1), Nucleoplasmin 2 (NPM2), Zygote arrest 1 (ZAR1), and Tripartite-motif protein-24 (TRIM24), using oocytes matured in 50 ng/ml, 100 ng/ml, or 1000 ng/ml of estradiol 17-β (E₂), 10% pFF, or protein-free maturation media. Expression of maternal effect genes, was shown by the ΔCt (cycle threshold) values, obtained from the difference between the Ct values of the normalizing gene (GAPDH) and the genes of interest evaluated through QRT-PCR. Values of ΔCt were analyzed in place of fold change to avoid data manipulation. The ΔCt expression of TRIM24 in 0 ng/ml E₂ maturation medium and the 10% pFF maturation medium were significantly different (p<0.05) from the non-matured control, the other maternal determinant genes did not differ in their expression under any treatment.en_US
dc.identifier.urihttp://hdl.handle.net/10388/etd-09142011-162338en_US
dc.language.isoen_USen_US
dc.subjectPorcineen_US
dc.subjectIn vitro Maturationen_US
dc.subjectMaternal Determinant gene expressionen_US
dc.subjectMicroarrayen_US
dc.subjectIn vitro Fertilizationen_US
dc.subjectOocyteen_US
dc.subjectEstradiolen_US
dc.titleInfluence of Estradiol on In vitro Maturation of Porcine Oocytesen_US
dc.type.genreThesisen_US
dc.type.materialtexten_US
thesis.degree.departmentVeterinary Biomedical Sciencesen_US
thesis.degree.disciplineVeterinary Biomedical Sciencesen_US
thesis.degree.grantorUniversity of Saskatchewanen_US
thesis.degree.levelMastersen_US
thesis.degree.nameMaster of Science (M.Sc.)en_US

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