"December, 2003" / Bibliography: leaves 153-161. / xii, 191, [20] leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / "The work presented in this thesis demonstrates that the exploitation of subtype-specific PDE inhibitors is a powerful experimental approach to study the oocyte and surrounding cumulus cells in separation, and to investigate the functions of cAMP in the two follicular compartments. The successful use of specific PDE isoenzyme inhibitors will prove important in the development of a clearer and more defined understanding of the fundamental mechanisms of regulating mammalian oocyte maturation." --p. 150. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 2004?

The ultimate purpose of this study was to investigate AHP2 protein function in Arabidopsis; AHP2 protein is known to form a heterodimer with MND1. But to do this an existing chromosome spreading protocol had to be modified to reproducibly provide large numbers of well preserved chromosomes and a multi-criteria meiotic staging method was developed to accurately identify chromosome spreads at specific prophase I substages. As well a technique for combined immuno-cytochemistry and fluorescent in situ hybridization (FISH) had to be developed. Coimmuno-localization of AHP2 and MND1 proteins, in wild type meiocytes, revealed synchronous temporal organization (signal initially peaks, for both, during zygotene) but spatially the AHP2 signal appeared exclusively as chromosome axis-associated foci whereas the MND1 signal was diffuse with some foci suggesting that MND1 may also be localizing to loop regions. This finding strongly suggests that MND1 also functions independently of AHP2 during meiosis. We coupled transmission electron microscopy (TEM) analysis of ultrathin sections of meiotic nuclei with light microscope (LM) analysis of chromosome spreads and demonstrated that ahp2 meiocytes fail to stabilize chromosome close alignment that normally occurs during zygotene. My method of combining 2-Bromo-5-deoxyUridine (BrdU) - determination of meiosis duration and the relative durations of each substage allowed us to calculate the absolute duration of each prophase I substage in wild type and revealed that early to mid-zygotene was prolonged in the ahp2 mutant. This finding was consistent with the ahp2 mutant’s overall lack of stabilized chromosome alignment. Scanning electron microscopy (SEM) showed that the Arabidopsis ahp2 short stamen filament was due to reduced cell elongation in filament parenchyma (epidermal) cells. Detection of illegitimate connections between chromosomes at anaphase I may trigger cell-to-cell signals that result in reduced epidermal cell elongation in the stamen filament. Finally, I report that the short arms of NOR-bearing chromosomes 2 and 4 homologously pair and synapse in the ahp2 mutant despite an overall lack of stabilized pairing. This NOR phenomenon has been observed in Drosophila but ours is the first report of the ability of NORs to locally induce pairing and synapsis in plants.

The ultimate purpose of this study was to investigate AHP2 protein function in Arabidopsis; AHP2 protein is known to form a heterodimer with MND1. But to do this an existing chromosome spreading protocol had to be modified to reproducibly provide large numbers of well preserved chromosomes and a multi-criteria meiotic staging method was developed to accurately identify chromosome spreads at specific prophase I substages. As well a technique for combined immuno-cytochemistry and fluorescent in situ hybridization (FISH) had to be developed. Coimmuno-localization of AHP2 and MND1 proteins, in wild type meiocytes, revealed synchronous temporal organization (signal initially peaks, for both, during zygotene) but spatially the AHP2 signal appeared exclusively as chromosome axis-associated foci whereas the MND1 signal was diffuse with some foci suggesting that MND1 may also be localizing to loop regions. This finding strongly suggests that MND1 also functions independently of AHP2 during meiosis. We coupled transmission electron microscopy (TEM) analysis of ultrathin sections of meiotic nuclei with light microscope (LM) analysis of chromosome spreads and demonstrated that ahp2 meiocytes fail to stabilize chromosome close alignment that normally occurs during zygotene. My method of combining 2-Bromo-5-deoxyUridine (BrdU) - determination of meiosis duration and the relative durations of each substage allowed us to calculate the absolute duration of each prophase I substage in wild type and revealed that early to mid-zygotene was prolonged in the ahp2 mutant. This finding was consistent with the ahp2 mutant’s overall lack of stabilized chromosome alignment. Scanning electron microscopy (SEM) showed that the Arabidopsis ahp2 short stamen filament was due to reduced cell elongation in filament parenchyma (epidermal) cells. Detection of illegitimate connections between chromosomes at anaphase I may trigger cell-to-cell signals that result in reduced epidermal cell elongation in the stamen filament. Finally, I report that the short arms of NOR-bearing chromosomes 2 and 4 homologously pair and synapse in the ahp2 mutant despite an overall lack of stabilized pairing. This NOR phenomenon has been observed in Drosophila but ours is the first report of the ability of NORs to locally induce pairing and synapsis in plants.

Thesis (Ph.D.)--York University, 2007. Graduate Programme in Biology. / Typescript. Includes bibliographical references (leaves 166-175). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&res_dat=xri:pqdiss&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft_dat=xri:pqdiss:NR39036

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Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1997. / Errata inserted. Bibliography: leaves 98-120.

Thesis supervisor: Robert E. Malone. Includes bibliographic references (p. 258-279).

Thesis (M.S.)--University of North Carolina at Greensboro, 2007. / Title from PDF title page screen. Advisor: John Tomkiel; submitted to the Dept. of Biology. Includes bibliographical references (p. 58-62).

Meiosis is the process by which sexually-reproducing organisms ensure that precisely half a chromosome set is passed from each parent to the following generation; this circumvents the doubling of the genome that would otherwise occur upon fertilisation. Meiosis occurs via a single round of DNA replication followed by two successive chromosome segregation events. In the first segregation, homologous chromosomes align and become physically linked through the process of meiotic recombination, which is crucial for the accurate segregation of homologous chromosomes. During the second round of segregation, sister chromatids are segregated to produce four haploid daughter cells. Failure to physically tether homologous chromosomes to each other through meiotic recombination can result in the aberrant segregation of homologous chromosomes, which can cause hereditary diseases (aneuploidies) and miscarriages in humans. Meiotic recombination also shuffles alleles of the parental chromosomes, which is crucial for evolution. The study of meiotic recombination, and its regulation, is thus paramount for our understanding of how genetic diversity is generated within populations. The work in this thesis has helped characterise factors, both genetic and environmental, that modulate meiotic recombination in the fission yeast, Schizosaccharomyces pombe. Here, I identify temperature as a major determinant of meiotic recombination outcome; when meiosis is performed at 16°C, significant reductions in meiotic recombination outcome are observed relative to meiosis performed at higher temperatures. Additionally, I present genetic and cytological evidence that the strand resection and strand invasion steps of meiotic recombination are impaired at 16°C relative to higher temperatures, but that double strand break levels appear not to be influenced by temperature. I have also characterised several novel genes predicted to be involved in meiotic recombination, and explored the genetic relationship between several genes already known to be crucial in modulating meiotic recombination. Finally, I have laid the foundations for a future project aiming to map the meiotic recombination landscape across the entire S. pombe genome.

Meiosis is a specialized cell division that produces haploid gametes from a diploid progenitor cell. It consists of one round of DNA replication followed by two consecutive rounds of chromosome segregation. Homologous chromosomes segregate in meiosis I and sister chromatids segregate in meiosis II. Failure to correctly regulate meiosis can result in aneuploidy, where daughter cells inherit an incorrect number of chromosomes. Aneuploidy is usually poorly tolerated in eukaryotes, and is associated with infertility, miscarriages and birth defects. At the meiosis I to meiosis II transition, DNA replication does not occur between chromosome segregation steps despite the need for Spindle Pole Bodies (SPBs) to be re-licensed in order to build meiosis II spindles. The mechanisms that make this distinction are not yet known. In budding yeast, the protein phosphatase Cdc14 is essential for the progression of cells into meiosis II. Cdc14 is sequestered for the majority of the cell cycle in the nucleolus by the inhibitor Cfi1/Net, and is only released in anaphase. We have observed Cdc14 localizing to and interacting with SPB components when nucleolar sequestration is inhibited. Through fluorescence microscopy and EM analysis, we have determined that Cdc14 is required for the re-duplication of SPBs after meiosis I. Our data implies a role for Cdc14 in the phospho-regulation of SPB half-bridge component Sfi1. Cdc14 is therefore essential for the relicensing of SPB duplication, a crucial step necessary to ensure accurate chromosome segregation in meiosis.