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Browsing by Author "Charlotte E. Farin, Committee Member"

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    Effects of Embryo Production Systems on Angiogenesis and Development of Bovine Placentas
    (2004-11-29) Miles, Jeremy R; Robert M. Petters, Committee Member; Jorge A. Piedrahita, Committee Member; Charlotte E. Farin, Committee Member; Peter W. Farin, Committee Chair; John E. Gadsby, Committee Co-Chair
    Placental abnormalities have been reported following the transfer of in vitro-produced (IVP) and cloned (somatic cell nuclear transfer; SCNT) embryos in cattle and sheep. The overall objective of this research was to determine the effects of embryo production systems (IVP and NT) on angiogenesis and development of placentas during early and late gestation in cattle. Angiogenesis was assessed by the expression of vascular endothelial growth factor (VEGF), peroxisome proliferator-activator receptor-gamma (PPARγ), and vascular morphometry. During late gestation (Day 222), angiogenesis and morphometry of placentas were assessed in placentas from embryos produced in vivo and in vitro using undefined, serum-supplemented medium (IVPS). During early gestation (Day 70), the effects of embryo culture media (undefined, IVPS or semi-defined modified synthetic oviductal fluid, mSOF) on angiogenesis and morphometry of placentas were evaluated. Finally, at Day 40 of gestation angiogenesis and development were compared in placentas from embryos produced in vivo, in vitro using G1.2/G2.2 media, or by SCNT. The results described in this dissertation demonstrated that angiogenesis as well as development of placentas differed depending on embryo production system and day of gestation. During late gestation, placentas from the IVPS group had decreased percentage of placentome surface area which was associated with increased expression of PPARγ protein and increased blood vessels-to-placentome surface area ratios. These findings suggest that during late gestation compensatory mechanisms exist in the vascular beds of placentas from bovine embryos produced in vitro. In contrast, during early gestation (Day 70) placentas from the IVPS group developed similar to in vivo controls; whereas placentas from the mSOF group had decreased densities of fetal and maternal blood vessels associated with a decreased expression of VEGF mRNA in cotyledons. At Day 40 of gestation, placentas from embryos produced using G1.2/G2.2 media appeared to have more compromised vascular development compared with placentas from embryos produced in vivo or by SCNT.
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    Final
    (2008-01-08) Sriperumbudur, Rajagopal; John E. Gadsby, Committee Chair; Charlotte E. Farin, Committee Member; William Flowers, Committee Member; Jorge A. Piedrahita, Committee Member
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    Production of Transgenic Chickens to Express Bacterial beta-Galactosidase and the Subsequent Utilization of Lactose as a Feed Stuff
    (2006-08-29) Borwornpinyo, Suparerk; James N. Petitte, Committee Chair; Paul E. Mozdziak, Committee Co-Chair; Samuel L. Pardue, Committee Member; Charlotte E. Farin, Committee Member
    The main objective of this dissertation was to create transgenic chickens expressing β-galactosidase as a genetic marker for cell lineage studies. To generate the transgenic chickens, the replication-defective retroviral-based SNTZ vector carrying the E. coli lacZ gene encoding nuclear-localized β-galactosidase was injected into subgerminal cavity of stage X (EG & K) White Leghorn embryos. Eight of 15 male chicks that survived to sexual maturity were germline chimeric birds based on PCR screening for the lacZ sequences in their semen. One of the eight lacZ-positive G0 roosters transmitted the lacZ gene to two male chicks from a total of 224 progeny (0.89%). From these two transgenic G1 males, the lacZ gene was stably transmitted through 4 generations in an expected Mendelian pattern for a single dominant allele. The expression of β-galactosidase was detected in cultured myoblasts derived from 1-d-old chick muscle, entire embryos, and in a variety of examined tissue types from young and adult chickens. In the current study, the generated transgenic lines which stably inherited and expressed the reporter lacZ gene are the first report of transgenic birds that could provide an alternative ideal cell marker for cell lineage studies. Qualitatively high expression of β-galactosidase was observed in villi of intestine. Potentially, the transgenic chickens that express β-galactosidase in the gastrointestinal tract could utilize lactose more efficiently. The second objective of this study was to determine whether the transgenic chickens can improve lactose digestibility and its use as an energy source. Overall, when dietary lactose was increased from 5 to 10%, the transgenic chickens showed lactose digestibility approximately 10% better than those of non-transgenic chickens. This is the first report of using gene transfer technology to manipulate the chicken genome to utilize feed more efficiently for agricultural purposes.

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