The cellular and molecular environment of the uterus during the pre - and peri - implantation period of early pregnancy is critical for implantation success and optimal fetal and placental development. Perturbations to this environment not only have consequences for the success of pregnancy and neonatal health and viability, but can also drive adverse health outcomes in the offspring after birth, particularly the development of metabolic disorders such as obesity, hypertension and insulin resistance. The influence of seminal plasma on the cytokine and immune uterine environment has been previously well characterised in mice, however the effects of disruption in uterine seminal plasma exposure for pregnancy outcome have not been investigated. The studies described in this thesis employed the use of surgical seminal vesicle ablation in males and embryo transfer experiments to investigate the physiological significance of uterine seminal plasma exposure on programming fetal and neonatal outcomes, and growth and metabolic status in adult offspring. We demonstrate that in the absence of seminal plasma, oocyte fertilisation and embryo implantation are reduced, showing that seminal plasma acts primarily to facilitate fertilisation, possibly by promoting sperm transport and survival in the reproductive tract. In addition we show that pregnancies initiated in the absence of seminal plasma give rise to offspring which display accelerated growth after birth and increased adiposity in adulthood, compared to those developed in a tract exposed to seminal plasma at the time of conception. Offspring conceived in the absence of seminal plasma also displayed alterations in serum leptin and adiponectin content, similar to those known to be associated with obesity in the mouse. Using embryo transfer experiments, we showed that some, but not all aspects of the perturbed postnatal development are recapitulated when embryos fertilised in the presence of what semen are transferred to a recipient tract which has not been exposed to seminal plasma. More severe perturbations were seen in 2 - cell transfer than in blastocyst transfer experiment. Additionally, there was a significant effect of the embryo transfer procedure, irrespective of seminal plasma exposure, on fetal and postnatal development that confounded interpretation of these experiments. In addition, we investigated the potential mechanisms by which the influence of seminal plasma is exerted. Mediators of pre - implantation embryo development, implantation and the modulation of the maternal immune response to pregnancy were all assessed for regulation by seminal plasma using QRT - PCR. It was demonstrated that seminal plasma exposure induces the up - regulation of key embryotrophic factors, LIF, GM - CSF and IL - 6, in the oviduct following insemination. Factors important in tissue remodelling required for implantation and angiogenesis, MMP - 2, MMP - 3 and VEGF - C, were also shown to be increased at the time of implantation after seminal plasma exposure. Additionally the generation of T - regulatory cells in uterine tissues, demonstrated by the up - regulation of the transcription factor FOXp3 was shown to be dependent on semen exposure. The influence of seminal plasma on embryonic development, implantation and modulation of the maternal immune response to pregnancy may therefore be mechanisms which contribute to the adverse outcomes seen in pregnancies initiated in the absence of seminal plasma. Together these experiments show a role for seminal plasma signalling at the time of insemination in influencing the pre - implantation embryo to program later fetal and neonatal development, thereby impacting on the metabolic health of offspring. We conclude that seminal plasma is not simply a transport medium for sperm, but acts also as a key regulator of a female tract environment providing optimal support for the developing embryo. / Thesis (Ph.D.)--School of Paediatrics and Reproductive Health, 2006.

Thesis (Ph. D.)--University OF MICHIGAN.

Thesis (M.A.) -- University of Texas at Arlington, 2008.

Title page, abstract and table of contents only. The complete thesis in print form is available from the University of Adelaide Library. / In mice and other mammalian species, deposition of semen into the female reproductive tract elicits a local inflammatory response. Whether a comparable response occurs within the human cervix has not previously been studied. The experiments described in this thesis demonstrate, using cervical tissue biopsies taken before and after intercourse, that exposure to semen elicits an infiltration of leukocytes into the cervical tissue of peri-ovulatory women. Immunohistochemical analysis identified macrophages and dendritic cells as the predominant leukocytes recruited into the cervical epithelium and stroma following intercourse. Cytotoxic / suppressor T lymphocytes and memory T cells were also increased. Comparable responses were not detected following condom-protected intercourse. Quantitative real-time PCR was performed on duplicate tissue biopsies to investigate the molecular regulation of this response. Expression of GM-CSF, a potent stimulator of myeloid cell recruitment, was found to increase by 2.5-fold following unprotected intercourse. Trends towards increased IL-6 and IL-8 mRNA were also observed. Condom-protected intercourse did not activate cytokine expression, further suggesting that exposure to semen, as opposed to mechanical trauma, provides the inflammatory stimulus. In an in vitro model using the immortalised Ect-1 cell line, TGFβ was identified as a candidate active seminal factor. All three TGFβ isoforms were capable of mimicking the stimulatory ability of seminal plasma in Ect-1 cells and were comparable in their capacity to stimulate both GM-CSF and IL-6 expression in a dose-responsive manner. The addition of TGFβ isoform-specific neutralising antibodies inhibited seminal plasma-induced increases in these cytokines. However TGFβ was unable to stimulate IL-8 production. Addition of IFNƴ was found to strongly inhibit TGFβ-stimulated GM-CSF production, and 19-0H PGE₁ was found to increase IL-6 and IL-8, but not GM-CSF production. Responses to seminal plasma constituents were almost exactly replicated in primary cultures of human ectocervical cells. These results identify TGFβ as the major active constituent in human seminal plasma and indicate that other seminal agents, 19-0H PGE₁ and IFNƴ, interact with TGFβ to differentially regulate cervical cytokine expression. Finally, whether human seminal plasma cytokine content was associated with fertility in men was examined. No relationship between seminal plasma TGFβ₁, TGFβ₂, TGFβ₃, IL-8 or bacterial endotoxin content and fertility status was observed. However, there was an increased likelihood of high IFNƴ content in the male partners of couples experiencing infertility, most notable in recurrent miscarriage. The discriminating value of IFNƴ was increased when evaluated as a ratio of total TGFβ content. Inflammatory changes after exposure of the female reproductive tract to seminal plasma are implicated in 'conditioning' the maternal immune response, to facilitate successful embryo implantation and pregnancy. The studies described in this thesis provide a mechanistic basis for the observations linking exposure to semen with pregnancy success in humans and have expanded our knowledge of the cellular and molecular events that occur within the female reproductive tract following intercourse. Seminal plasma can therefore no longer be thought of as merely a transport medium for spermatozoa, rather as a means for communication between the male and female reproductive tissues, potentially required for optimal pregnancy success. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1213820 / Thesis (Ph.D.) -- University of Adelaide, Medical School, 2005

Title page, abstract and table of contents only. The complete thesis in print form is available from the University of Adelaide Library. / In mice and other mammalian species, deposition of semen into the female reproductive tract elicits a local inflammatory response. Whether a comparable response occurs within the human cervix has not previously been studied. The experiments described in this thesis demonstrate, using cervical tissue biopsies taken before and after intercourse, that exposure to semen elicits an infiltration of leukocytes into the cervical tissue of peri-ovulatory women. Immunohistochemical analysis identified macrophages and dendritic cells as the predominant leukocytes recruited into the cervical epithelium and stroma following intercourse. Cytotoxic / suppressor T lymphocytes and memory T cells were also increased. Comparable responses were not detected following condom-protected intercourse. Quantitative real-time PCR was performed on duplicate tissue biopsies to investigate the molecular regulation of this response. Expression of GM-CSF, a potent stimulator of myeloid cell recruitment, was found to increase by 2.5-fold following unprotected intercourse. Trends towards increased IL-6 and IL-8 mRNA were also observed. Condom-protected intercourse did not activate cytokine expression, further suggesting that exposure to semen, as opposed to mechanical trauma, provides the inflammatory stimulus. In an in vitro model using the immortalised Ect-1 cell line, TGFβ was identified as a candidate active seminal factor. All three TGFβ isoforms were capable of mimicking the stimulatory ability of seminal plasma in Ect-1 cells and were comparable in their capacity to stimulate both GM-CSF and IL-6 expression in a dose-responsive manner. The addition of TGFβ isoform-specific neutralising antibodies inhibited seminal plasma-induced increases in these cytokines. However TGFβ was unable to stimulate IL-8 production. Addition of IFNƴ was found to strongly inhibit TGFβ-stimulated GM-CSF production, and 19-0H PGE₁ was found to increase IL-6 and IL-8, but not GM-CSF production. Responses to seminal plasma constituents were almost exactly replicated in primary cultures of human ectocervical cells. These results identify TGFβ as the major active constituent in human seminal plasma and indicate that other seminal agents, 19-0H PGE₁ and IFNƴ, interact with TGFβ to differentially regulate cervical cytokine expression. Finally, whether human seminal plasma cytokine content was associated with fertility in men was examined. No relationship between seminal plasma TGFβ₁, TGFβ₂, TGFβ₃, IL-8 or bacterial endotoxin content and fertility status was observed. However, there was an increased likelihood of high IFNƴ content in the male partners of couples experiencing infertility, most notable in recurrent miscarriage. The discriminating value of IFNƴ was increased when evaluated as a ratio of total TGFβ content. Inflammatory changes after exposure of the female reproductive tract to seminal plasma are implicated in 'conditioning' the maternal immune response, to facilitate successful embryo implantation and pregnancy. The studies described in this thesis provide a mechanistic basis for the observations linking exposure to semen with pregnancy success in humans and have expanded our knowledge of the cellular and molecular events that occur within the female reproductive tract following intercourse. Seminal plasma can therefore no longer be thought of as merely a transport medium for spermatozoa, rather as a means for communication between the male and female reproductive tissues, potentially required for optimal pregnancy success. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1213820 / Thesis (Ph.D.) -- University of Adelaide, Medical School, 2005

Title page, abstract and table of contents only. The complete thesis in print form is available from the University of Adelaide Library. / In mice and other mammalian species, deposition of semen into the female reproductive tract elicits a local inflammatory response. Whether a comparable response occurs within the human cervix has not previously been studied. The experiments described in this thesis demonstrate, using cervical tissue biopsies taken before and after intercourse, that exposure to semen elicits an infiltration of leukocytes into the cervical tissue of peri-ovulatory women. Immunohistochemical analysis identified macrophages and dendritic cells as the predominant leukocytes recruited into the cervical epithelium and stroma following intercourse. Cytotoxic / suppressor T lymphocytes and memory T cells were also increased. Comparable responses were not detected following condom-protected intercourse. Quantitative real-time PCR was performed on duplicate tissue biopsies to investigate the molecular regulation of this response. Expression of GM-CSF, a potent stimulator of myeloid cell recruitment, was found to increase by 2.5-fold following unprotected intercourse. Trends towards increased IL-6 and IL-8 mRNA were also observed. Condom-protected intercourse did not activate cytokine expression, further suggesting that exposure to semen, as opposed to mechanical trauma, provides the inflammatory stimulus. In an in vitro model using the immortalised Ect-1 cell line, TGFβ was identified as a candidate active seminal factor. All three TGFβ isoforms were capable of mimicking the stimulatory ability of seminal plasma in Ect-1 cells and were comparable in their capacity to stimulate both GM-CSF and IL-6 expression in a dose-responsive manner. The addition of TGFβ isoform-specific neutralising antibodies inhibited seminal plasma-induced increases in these cytokines. However TGFβ was unable to stimulate IL-8 production. Addition of IFNƴ was found to strongly inhibit TGFβ-stimulated GM-CSF production, and 19-0H PGE₁ was found to increase IL-6 and IL-8, but not GM-CSF production. Responses to seminal plasma constituents were almost exactly replicated in primary cultures of human ectocervical cells. These results identify TGFβ as the major active constituent in human seminal plasma and indicate that other seminal agents, 19-0H PGE₁ and IFNƴ, interact with TGFβ to differentially regulate cervical cytokine expression. Finally, whether human seminal plasma cytokine content was associated with fertility in men was examined. No relationship between seminal plasma TGFβ₁, TGFβ₂, TGFβ₃, IL-8 or bacterial endotoxin content and fertility status was observed. However, there was an increased likelihood of high IFNƴ content in the male partners of couples experiencing infertility, most notable in recurrent miscarriage. The discriminating value of IFNƴ was increased when evaluated as a ratio of total TGFβ content. Inflammatory changes after exposure of the female reproductive tract to seminal plasma are implicated in 'conditioning' the maternal immune response, to facilitate successful embryo implantation and pregnancy. The studies described in this thesis provide a mechanistic basis for the observations linking exposure to semen with pregnancy success in humans and have expanded our knowledge of the cellular and molecular events that occur within the female reproductive tract following intercourse. Seminal plasma can therefore no longer be thought of as merely a transport medium for spermatozoa, rather as a means for communication between the male and female reproductive tissues, potentially required for optimal pregnancy success. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1213820 / Thesis (Ph.D.) -- University of Adelaide, Medical School, 2005

Thesis (Ph. D.)--University of New South Wales, 2002. / Also available online.

Thesis (Ph. D.)--University OF MICHIGAN.

Thesis (Ph. D.)--University of Missouri-Columbia, 2000. / Typescript. Vita. Includes bibliographical references (leaves 137-162). Also available on the Internet.

Thesis (doctoral)--Joensuun yliopisto, 2005. / Added t.p. and errata slip for series numbering from publisher inserted. Includes bibliographical references (p. 397-430) and index.