Browsing by Subject "gene"
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Item Effects of exercise or oocyte heat shock on embryo development and gene expression in the horse(2009-05-15) Mortensen, Christopher JohnHorse owners commonly maintain their broodmares in training and competition during the breeding season. The effect this has on mare reproductive efficiency has received limited attention. Heat stress has shown to be detrimental to oocyte competence in other species and heat shock protein 70 has been shown to be an important gene in regulating cellular response to heat. Mares were exercised in a hot humid environment to determine the effects on reproductive efficiency. Embryos were collected at d 7 after ovulation from exercised and control mares. Oocyte developmental competence was measured after oocytes were subjected to a one time heat shock, 42 ?C for 2 or 4 h, at the onset or near completion of in vitro maturation. Embryos from both previous experiments were examined for HSP70 gene expression by real time RT-PCR. Exercised mares ovulated significantly smaller follicles, 39.8 vs. 41.5 mm diameter, and ovulated later after being given PGF2?, 8.5 vs. 9.2 d. Twenty-two embryos (22/35) were recovered from control mares, recovery rate of 63%. Significantly fewer embryos were recovered in exercised mares (11/32), recovery rate of 34%. A lower proportion of grade 1 embryos were recovered from exercised versus control mares (4/11 vs.16/22,respectively). No effect was observed on oocyte nuclear maturation or embryonic development after ICSI when oocytes were exposed to heat shock at the onset of IVM. A heat shock of 42 ?C for 2 or 4 h on oocytes during late IVM resulted, however, in a significantly lower rate of nuclear maturation, and a significant decrease in advanced embryo development (morulae plus blastocysts). Heat shock protein 70 gene expression was shown to be related to quality score of in vivo-recovered embryos, with lower quality embryos recording a significantly higher relative expression. Heat shock of late stage IVM oocytes for 4 h resulted in significantly higher blastocyst HSP70 expression. Results of this study indicate that exercise in a hot humid environment is detrimental to mare reproductive efficiency, late-stage maturing oocytes are sensitive to heat, and HSP70 expression in equine embryos is related to embryo quality score and oocyte quality.Item Genomic analysis of sorghum by fluorescence in situ hybridization(Texas A&M University, 2004-11-15) Kim, Jeong-SoonThe reliability of genome analysis and proficiency of genetic manipulation in vivo and in vitro are increased by assignment of linkage groups to specific chromosomes, placement of centromeres, orientation with respect to telomeres, and linear alignment with respect to chromosomal features and dimensions. I undertook five studies aimed at integrating sorghum genomics and cytogenetics at several levels. The results help establish an entirely new "cyto-genomics" resource, impacts of which are likely to be broad. In the first study, I developed a FISH-based karyotyping system for Sorghum bicolor Moench. I used integrated structural genomic resources, including linkage maps and large-insert clonal libraries of sorghum genomic DNA to develop a 17-locus probe cocktail for simultaneous fluorescent in situ hybridization (FISH). This probe enabled facile identification of all chromosome pairs in mitotic chromosome spreads. Perhaps just as important, I established time-efficient means to select sorghum BAC clones for multi-probe FISH. Thus, an integrated cyto-genomics system for sorghum can be constructed without need of chromosome flow sorting or microdissection, both of which are difficult and costly. In the second study, hybridization of DNA clones from 37 different genomic regions enabled the assignment of linkage groups and orientation of linkage maps to chromosomes. Comparisons between genetic and physical distances throughout the genome enabled a new nomenclature for linkage group designation in sorghum. The results provide an integrated nomenclature system of Sorghum bicolor chromosomes and linkage groups. In the third study, I created high-resolution maps by FISH to pachytene bivalents for two linkage groups (B and H), and defined relationships between pericentromeric heterochromatin, centromeres, mapped markers and recombination rates. These relationships will help guide the development and use of sorghum genomics. In the fifth study, I used FISH in two ongoing gene-targeted efforts. For the maturity gene ma5 and fertility restoration gene rfl, I estimated physical lengths between currently available flanking molecular markers. This enables estimation of recombination densities in these regions and assessment of the applicability of map-based and -assisted cloning.