Please read the abstract in the section 00front of this document / Dissertation (MSc)--University of Pretoria, 2005. / Zoology and Entomology / Unrestricted

published_or_final_version / abstract / Anatomy / Doctoral / Doctor of Philosophy

Conclusion. Taken together, paternal factors carried in ASG secretion affect genomic imprinting of developing embryos. The outcome of research work described here deepens our understanding of the role of ASG in maximizing reproductive performance mediated by regulating the epigenetic marks of the genome and in particular the imprinted genes. / Introduction. Our previous in vivo studies in golden hamster have shown the accessory sex glands (ASG) secretion facilitate the development of embryos to term but the underlying mechanism is still not clear. Since the deleterious effect caused by the lack of sperm exposure to ASG secretion is heritable to developing fetus and even after birth, we hypothesized that the paternal factor carried in ASG secretion may change the epigenetic regulation and in particular the imprinted genes of embryonic genome. / Materials and methods. Golden hamster and ICR mouse were used in this study. Hamster is a well-established animal model to study the effect of individual ASG but the genetic background of hamster is poorly known. To verify the specificity of our molecular probe and antibodies used in hamster, a mouse model was also established. Five groups of male hamsters and two groups of male mice were established by surgical treatment. In hamster, (SH) sham-operated, (VPX) ventral prostate-removed, (TX) all ASG-removed, (VPVX) castrated with ASG-removed except ventral prostate and (VX) castrated with intact ASG were established. In mouse, SH and VPX were established. In single-mating of hamster, male was copulated with female at estrus for 15 min. In double-mating of hamsters, female mated with each male for 10 min each. In single-mating of mouse, male was caged with female for 1 h. Epididymal sperm, uterine sperm, fertilized oocytes, pre-implantation embryos and fetuses at 13 days gestation (E13) were collected. Global DNA methylation of sperm, fertilized oocytes, early embryos and E13 fetuses were investigated by indirect immunofluorescence and DNA dot-blot using antibody against methylated DNA. Using the same technique, histone acetylation at lysine 5 residue was detected in male pronuclei of fertilized oocytes, protamine 1 and 2 content were detected in sperm, DNA methyltransferase 1, 3a and 3b activities were detected in early embryos. The crown-rump length and weight of fetuses were measured. Morphology was also examined under scanning electron microscope. Two sets of co-ordinately regulated but oppositely expressed imprinted genes Igf2/H19 and Dlk1/Gtl2 were investigated. H19 differentially methylated region (DMR) and Gtl2 promoter were examined by bisulfite sequencing in sperm and E13 fetuses. Expression of Igf2 and Dlk1 were examined by in situ hybridization and real-time PCR in pre-implantation embryos and E13 fetuses. / Results. Uterine sperm in VPX and TX groups showed no change of DNA methylation level and protamine 1 and 2 content. Fertilized oocytes in VPX and TX groups showed similar DNA methylation level as SH group in both hamster and mouse. Histone hypoacetylation was observed in male pronuclei of hamster but not in mouse. Early embryos in VPX and TX groups showed abnormal level of DNA methylation and Dnmt3b during embryo development in hamster. Replenishment of ASG secretion to sperm from VPX and TX group by double-mating restored the DNA methylation level to normal in early embryos. E13 fetuses of VPX and TX groups in hamster and VPX group in mouse showed DNA hypomethylation. E13 fetuses of VPX group in hamster showed increase in average crown-rump length and body weight with larger variations between individuals. One E13 fetus of VPX group in mouse showed polydactyly and malformation in the head. Real-time PCR showed abnormal expression of Igf2 and Dlk1 in both pre-implantation embryos and E13 fetuses of VPX and TX groups. Bisulfite sequencing showed hypermethylation of H19 DMR in VPX and TX groups of hamster and hypomethylation of Gtl2 promoter in VPX group of mouse. / "August 2007." / Adviser: Pak Ham Chow. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4739. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 194-224). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

Poor semen quality is an indicator of male infertility and is also associated with a variety of adverse health outcomes in men. It is therefore important from both clinical and public health perspectives to discover predictors of poor semen quality, especially those that are potentially modifiable. My dissertation research focuses on two of these potential risk factors: adiposity and stress. Unlike most studies to date, which have only considered the relationship between these exposures and semen quality cross-sectionally, my research takes a life course approach. I explore associations between birth weight, adiposity in both childhood and adulthood, and allostatic load—a construct representing the effect of cumulative stress on the body’s regulatory systems—and three commonly-used semen outcome parameters: sperm concentration, percent progressive motility, and percent normal morphology. The logic that underlies this approach is that while sperm are constantly being produced from sperm stem cells in the testes, which would argue in favor of cross-sectional studies, the sperm stem cells themselves and the Sertoli and Leydig cells that stimulate and nurture that metamorphosis are laid down in the fetal period and undergo important developmental and proliferative phases during early childhood and puberty that affect their number and functional health in adulthood. Using data from 193 participants in the Study of the Environment and Reproductive health follow-up to the Child Health and Development Studies birth cohort, I was able to calculate birth weight for gestational age percentile (bw/ga) and six age-appropriate adiposity measures (at 4 months, 12 months, and 4 years, and in participants’ 20s, 30s, and at the time of semen collection), then test for their independent, critical period, and cumulative associations with the three semen outcomes as well as a combined outcome measure of subfertility based on World Health Organization reference levels. While bw/ga was positively associated with sperm concentration, subsequent childhood adiposity measures showed increasingly negative associations, and none of the adult measures were significantly associated with concentration. By contrast, only adult measures were associated with motility and morphology. This suggests that there may be critical periods in childhood when adiposity negatively affects sperm concentration by interfering with the development and proliferation of Sertoli and Leydig cells. Accumulation of oxidative stress in the testes due to overweight/obesity may explain the negative relationships between adult adiposity and sperm motility and morphology. To investigate allostatic load’s relationship to semen quality, I conducted a pilot study at Columbia University’s Center for Women’s Reproductive Care that enrolled 61 men who were having their initial diagnostic semen analysis and blood draw on the same day. Blood samples were analyzed for 7 biomarkers associated with homeostatic regulation across several physiologic domains. I then created an allostatic load scale in which participants were assigned 1 point for being in the high-risk quartile for systolic blood pressure, diastolic blood pressure, body mass index, or any of the biomarkers. In regression analyses, allostatic load was not associated with either sperm concentration or morphology, but showed an unexpected positive association with motility. This association was entirely driven by the six participants who scored 0 on the allostatic load scale and who did not differ from the rest of the sample in any way that could plausibly be linked to reduced motility. I therefore concluded that this was a spurious finding. In further analysis of the allostatic load variable itself, I found that few of its individual components were correlated with the semen outcomes. This contrasts with other studies of allostatic load and adverse health outcomes, but these have generally been conducted in either elderly or stressed populations, neither of which described my cohort. Allostatic load may not be a reliable measure of stress in reproductive age populations and may not capture regulatory systems appropriate to reproductive health outcomes. My dissertation highlights the value and challenges of conducting semen quality research from a life course perspective. Future studies should consider collecting longitudinal data on adiposity and stress, as well as repeated semen samples beginning in adolescence in order to further our understanding of the natural progression of semen quality across the reproductive life span and provide the opportunity to explore whether modifying these risk factors affects semen quality.

Cheung King Ho. / "August 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 140-158). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Chan Oi Chi. / "May 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 259-310) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

"April 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 158-181). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Parmi les différents types de canaux ioniques, les canaux potassium (K+) sont très largement exprimés au niveau des cellules eucaryotes. Ils se répartissent en plusieurs familles et sous-familles. Parmi celles-ci, les canaux K+ ATP-dépendants (KATP) représentent une classe tout à fait particulière. En effet, ils ont la particularité d’être sensibles à la concentration cytosolique d’ATP et permettent ainsi de coupler le potentiel membranaire de la cellule à son statut métabolique.<p>Le canal KATP est un complexe hétéro-octamérique constitué de 2 sous-unités :une sous-unité Kir6.x (Kir6.1 ou Kir6.2) formant le pore du canal et appartenant à la famille des canaux potassiques de type « inward rectifier », et une sous-unité régulatrice SURx (SUR 1 ou SUR2A/B) faisant partie des protéines ABC (ATP-binding cassette). L’expression hétérologue des sous-unités Kir6.x et SURx suivant différentes combinaisons conduit à la formation de plusieurs types de canaux KATP possédant des propriétés électro-physiologiques et des sensibilités aux nucléotides et aux agents pharmacologiques distinctes. \ / Doctorat en sciences biomédicales / info:eu-repo/semantics/nonPublished