Thesis (Ph.D.)--Boston University / A number of biologically active compounds known to influence various aspects of mammalian reproduction were investigated in order to determine specifically whether or not they affect gonadotrophin-induced ovulation and, if so, whether they act directly on the ovaries by blockage of the utilization of gonadotrophins or indirectly via the hypothalamico-hypophyseal axis. Maximal ovulatory response of 35 +/- 5 ova were routinely induced in 21-day-old intact Wistar rats by the subcutaneous injection of 20 IU pregnant mare serum (PMS) followed by the intraperitioneal injection of 25 IU human chorionic gonadotrophin (HCG) 56 hours later. Animals were sacrificed 20 hours after HCG injection, and body weights were recorded. Fallopian tubes were removed, and ova within them were recovered and counted. Ovaries were weighed to the nearest 0.2 mg and weights expressed as actual wet weight and relative weight (mg/100 gm body weight). Average numbers of ova shed and ovarian weights of the gonadotrophin-treated control groups were compared with those of the experimental groups, which were also administered single doses of the biologically active compounds. If a given compound significantly inhibited the superovulatory response in intact rats, it was then administered under the same conditions to rats hypophysectomized at 25 days of age. [TRUNCATED]

The Sertoli cell tight junction (TJ) of the seminiferous epithelium is important for the developmental process of spermatogenesis as it separates germ cells in the seminiferous tubules from the general circulation in the testicular interstitium. Absence of the TJ leads to spermatogenic arrest and infertility. TJs form at puberty as circulating gonadotrophins luteinising hormone/testosterone and follicle stimulating hormone increase. Several studies have demonstrated hormonal regulation of the two major TJ proteins, claudin-11 and occludin, and also of TJ function in vitro and in vivo. Men with low levels of circulating gonadotrophins exhibit an immature and dysfunctional TJ phenotype, which is reversed upon the exogenous application of gonadotrophins. This thesis hypothesises that claudin-11 and occludin are the major contributors to TJ function, and that gonadotrophins regulate TJ function and structure via these two proteins in several species including humans. This PhD was divided into four separate studies to address these hypotheses. The first study selectively silenced the genetic expression of claudin-11 and occludin with small interfering RNA (siRNA) in cultured immature rat Sertoli cells to determine their contribution to Sertoli cell TJ function in vitro. siRNA treatment against either protein significantly (p less than 0.01) reduced TJ function by ~50% as assessed by transepithelial electrical resistance. Immunocytochemistry displayed marked reductions in the localisation of these proteins to the TJ after siRNA treatment. It was concluded that both proteins significantly contributed to TJ function in vitro. The second and third studies then aimed to study hormonal regulation of the TJ in vivo. Weekly injections of the gonadotrophin releasing hormone antagonist acyline were used to suppress circulating gonadotrophins and spermatogenesis in adult rats. Acyline treatment disrupted i) the localisation of occludin to the TJ and ii) TJ function as shown by permeability to a biotin tracer, which was impermeable to TJs in controls. Short-term hormone replacement partially restored the effects of gonadotrophin suppression. It was concluded that gonadotrophins regulate the maintenance of the TJ in rats in vivo. The third study used the hypogonadal (hpg) mouse, which is a naturally occurring model of gonadotrophin deficiency with inactive spermatogenesis. Claudin-11 in hpg mice was not localised at the TJs, and these were dysfunctional as shown by permeability to biotin. Following hormone treatment, TJs were structurally and functionally competent, demonstrating that gonadotrophins also regulate the formation of TJs in vivo. The fourth study subsequently analysed TJs in gonadotrophin suppressed men, and it was found that claudin-11 staining was reduced from continuous bands in control men, to punctate staining in gonadotrophin-suppressed men, demonstrating that gonadotrophins also regulate the localisation of claudin-11 to the TJ in men in vivo. In summary, it is concluded that the Sertoli cell TJ is hormonally regulated, and that the major contributors to TJ function in vivo and in vitro are claudin-11 and occludin. It is hypothesised that the reduction of claudin-11 localisation to the TJ in men may also result in a loss of human Sertoli cell TJ function, suggesting that the TJ may be a potential target of hormonal contraception in men.

Almost a century has elapsed since the antigen-antibody interaction was first recognised as the basis of an immune response (Ehrlich, 1897). However, it was only in the 1930s, with the development of improved technologies that this concept was better understood, and led to the discovery of the amazing diversity and specificity of antibody molecules (Landsteiner, 1933). Theoretically, it is possible to make antibodies to a variety of biological substances and other chemicals, and therefore they are ideally suited as specific recognition elements to be used for analytical, cytological, functional, therapeutic and biochemical purposes. The development of the radioimmunoassay (RIA) thirty five years ago, revolutionised research in many areas of clinical and scientific investigation. This technique evolved rapidly from the discovery made by Berson et al. in 1956 that antibodies to insulin could be detected in patients treated with this hormone, by measuring the binding of radiolabeled insulin to these antibodies. Although in the past RIAs have been the most important assay system employing antibody and labelled tracer, the limitation was that reliance had to be placed on the chance development of a good polyclonal antibody. These shortcomings stimulated the search for monospecific antibodies of reproducible quality and sufficient quantity. The development and introduction of monoclonal antibody technology brought about a revolution in immune serology (Kohler and Milstein, 1975). Establishment of immortal cell lines which contained the genetic elements of antibody-producing cells was achieved by fusion between a myeloma cell line and spleen cells from an immunised donor. The resulting hybrids had the essential properties of both parents, namely, permanent growth and a high capacity for the synthesis and secretion of immunoglobulins, normally characteristics of plasmacytomas, together with the genetic elements defining a specific antibody. Gestational trophoblastic disease (GTD) is a neoplastic condition of the trophoblast and occurs as molar pregnancy in a benign or invasive form, or as choriocarcinoma in a malignant form. Effective therapy has been developed for the treatment of both choriocarcinoma and molar pregnancy, but the key to successful management of these patients lies in their prompt diagnosis and careful monitoring of response to treatment (Green-Thompson, 1986). Fortuitously, these tumours elaborate the human chorionic gonadotrophin hormone (hCG) and its free alpha (a) and beta (B) subunits and hence a ready marker for the tumour exists. Human chorionic gonadotrophin is one of a group of glycoprotein hormones, which includes luteinising hormone (LH), follicle stimulating hormone (FSH) and thyroid stimulating hormone (TSH). These hormones are composed of two dissimilar subunits designated a and B, which are bound non-covalently in the intact molecule. The B-subunit of each glycoprotein hormone is unique and is responsible for the respective biological and immunological properties of the glycoproteins. In contrast, all four hormones possess an identical a-subunit which is coded for by a single gene (Fiddes and Goodman, 1979). The measurement of hCG and its free B-subunit, as so-called BhCG, for the diagnosis and monitoring of therapy in patients with GTD is now routinely practised throughout the world (Vaitukaitis et al., 1972). However it has been demonstrated by Bagshawe (1975) that when serum BhCG can no longer be measured by current RIA methods, up to 10" tumour cells may remain undetected. In addition, there have been isolated reports of two patients with choriocarcinoma in whom BhCG was undetectable in the serum but who appeared to be secreting only the a-subunit (Dawood et al, 1977). Furthermore, it has been suggested that measurement of free a-subunit rather than intact hCG or the free B-subunit is a more effective means of detecting persistent trophoblastic disease as well as tumour recurrence following treatment (Quigley et al, 1980a and b). Radioimmunoassays which measure the free a-subunit of hCG have been developed, but in general lack the specificity and sensitivity required (Gaspard et al, 1980; Kohorn et al, 1981). These assays employ polyclonal antisera which also detect epitopes common to the pituitary gonadotrophins. Thus there is a need to produce monoclonal antibodies which recognise regions of the free a-subunit which are hidden in the intact gonadotrophins. Such antibodies would provide the required specificity for use in RIAs but are limited in their use by their inherent lack of high affinity for the antigen. Fortunately, this drawback may be overcome by using monoclonal antibodies as labelled reagents in an alternative assay system, the immunoradiometric assay (IRMA), described by Miles and Hales (1968). The IRMA, particularly the two-site sandwich version of the assay, has been shown to provide greater sensitivity in addition to allowing enhanced specificity. This is a consequence of the use of two antibodies in excess to detect the analyte, each directed at a different epitope on the target molecule. The first antibody, referred to as the capture antibody, is usually linked to a solid-phase to facilitate easy separation and is added in excess relative to the target hormone to enhance antibody-antigen interaction, thereby allowing increased sensitivity in the measurement of analyte. The second antibody, referred to as the detection antibody, is labelled with a radioactive isotope or an enzyme to detect antigen already bound to the capture antibody. The application of monoclonal antibodies specific for the free a-subunit to a highly sensitive IRMA format is an obvious need. Hence this study was undertaken firstly, to raise and characterise monoclonal antibodies to the free a-subunit, secondly to develop an IRMA using these antibodies and finally to establish whether measurement of free a-subunit has any clinical advantage. / Thesis (M.Med.Sc.)-University of Natal, Durban, 1990.

The layer of cells lining the outer surface of the mammalian ovary, the ovarian surface epithelium (OSE), is a constant feature throughout the dynamic tissue remodeling that occurs throughout the reproductive cycle (follicle growth, ovulation, corpora lutea formation and pregnancy). Abnormal development of these cells is responsible for 90% of all epithelial ovarian cancers in women and epidemiological studies have shown that susceptibility to ovarian cancer is negatively correlated with increasing pregnancy. Little is known about how OSE cells are affected at each stage of the cycle, so the main aim of this study was to determine how the reproductive cycle affected proliferation and degeneration of OSE cells. This study utilised three animal models each with a different type of reproductive cycle: a mono-ovular seasonal breeder (Sheep), a mono-ovular polyoestrous breeder (Cow) and a poly-ovular non human primate (marmoset) to allow comparisons to be made. Comparison of OSE proliferative activity was made in sheep and marmoset at each stage of the cycle including pregnancy and anoestrous. The bovine model was used to investigate apoptotic cell death. Proliferative activity of somatic cells within the sheep ovary was monitored throughout the reproductive cycle by detection of cell cycle markers PCNA and Ki67 using immunohistochemistry. The pattern of OSE proliferation was correlated with the pattern of follicle development at each stage (sheep and marmoset). During pregnancy cell proliferation was significantly lower in OSE and in granulosa cells, reflecting a suppression of mature follicle development during these stages whereas in cycling animals proliferation was increased. Differences in OSE proliferation were observed in relation to the local underlying tissue environment in both sheep and marmoset. Epithelial cell rupture and regeneration enhanced the hormonal mitogenic action on epithelial cells, which showed highest proliferation over corpora lutea in each animal model. To test the hypothesis that these changes are mediated by hormones or growth factors ovine OSE cells were cultured and proliferative activity monitored after treatment with several factors: fetal calf serum (FCS), follicular fluid from follicles of varying sizes, corpora lutea extracts, recombinant human IGF-1, oestradiol and progesterone. IGF alone was demonstrated to have an affect on increasing proliferation of cultured OSE cells. Levels of FSHr and LHr were monitored by quantitative real- time PCR and it was demonstrated that the concentration of gonadotrophin receptors in OSE, increased prior to and after ovulation, at which time the in vivo OSE proliferation also peaked. The in situ apoptosis index was determined in bovine tissue using TUNEL throughout the regular cycle, and at mid and late-pregnancy stages. The results showed that pregnancy induced apoptotic activity in OSE cells and up regulated the tumour suppressor gene p53. Cultured bovine OSE cells also exhibited an increased level of apoptosis following progesterone treatment. Since p53/p53 gene expression in OSE over the corpora lutea producing progesterone also increased, this progesterone-mediated apoptosis may be mediated through an up-regulation of p53 synthesis. The effect of pregnancy and low production of gonadotrophins in the regulation of OSE cell morphology and activity was further investigated in the marmoset monkey (a non-human primate) treated with GnRH antagonist and infused with BrdU to monitor proliferative activity. OSE proliferation was correlated to ovarian events (follicular growth, ovulation and luteinization) and this was suppressed during pregnancy. Inhibition of gonadotrophin secretion by treatment with a GnRH antagonist also markedly inhibited OSE proliferation. Taken together these studies support the hypothesis that pregnancy and periods of anovulation reduce proliferation of OSE cells and alter the pattern of apoptotic cell death and that this effect is independent of species and reproductive pattern. Suppression of gonadotrophins and other growth factors during pregnancy could enhance p53-mediated apoptosis of damaged and mitogenic cells arising from repeated ovulations. This effect may partly explain why increasing number of pregnancies in woman reduces the chance of epithelial ovarian cancers.

O estudo objetivou avaliar num programa biotécnico aplicado a uma granja comercial, os feitos combinados, emprego de gonadotrofinas e aplicação do \"flushing\". As gonadotrofinas empregadas na indução e sincronização do estro à puberdade em marrãs, aliado a ciclicidade até o 4º estro, correspondeu à aplicação ou não da combinação hormonal, 600 UI de eCG (Novormon®, Syntex S. A., Argentina) e após 72 horas, 2,5mg de LH porcino (Lutropin®, Vetrepharm Canadá Inc., Canadá). O \"flushing\" alimentar ou esquema alimentar adotado na granja, foi empregado no ciclo estral que antecedeu a primeira inseminação artificial, ocorrida aos 220 dias de idade das fêmeas. Os parâmetros analisados foram: taxa de concepção (TC), taxa de parto (TP), taxa de aproveitamento (TA) e tamanho de leitegada (TL), sendo esse último representado pelo número de leitões nascidos totais (NT), nascidos vivos (NV), natimortos (NM) e mumificados (MM). Foram utilizadas 119 leitoas híbridas, num delineamento experimental inteiramente casualizado em arranjo fatorial 2x2. O tratamento com hormônio mostrou percentual significativamente maior de fêmeas que manifestaram estro até o 5° dia de início da indução, em comparação com o tratamento somente com o estímulo do macho, 18,33% vs. 5,08%, respectivamente (P=0,0249). No intervalo seguinte de 18 - 30 dias após a indução, considerando variação do ciclo estral de 18 a 25 dias, a combinação hormonal revelou percentual significativamente maior comparado com o estímulo natural do macho, 48,33% vs. 16,94%, respectivamente (P=0,0003). Nos intervalos subseqüentes as diferenças não foram significativas. Não foi detectada interação significativa para os efeitos combinados gonadotrofinas e \"flushing\". Na análise das características em separado, não houve diferença significativa na taxa de concepção para os tratamentos combinação hormonal e estímulo do macho (96,23% vs. 94,00%, respectivamente), e tratamentos com \"flushing\" e esquema alimentar da granja (95,92% vs. 94,44%, respectivamente). Na taxa de parto, os percentuais embora não tenham revelado significância, mostraram diferenças numéricas, sendo o maior valor numérico representado pelo tratamento com hormônio em comparação com o macho, 92,45% vs. 80,00% respectivamente (P=0,0653). Quanto à taxa de aproveitamento das fêmeas, considerando a indução aos 153 dias de idade das marrãs até o parto, da mesma maneira evidenciou-se diferença numérica mostrando o tratamento com hormônio percentual maior em comparação com macho, 81,67% vs. 67,80%, respectivamente (P=0,0815). Não houve diferença significativa nos tratamentos relacionados à aplicação do \"flushing\" e esquema alimentar da granja, 70,00% vs. 79,66%, respectivamente. Não foi evidenciada diferença significativa quanto ao total de nascidos, nascidos vivos, natimortos e mumificados, para os dois fatores. Destacou-se diferença numérica de 0,75 leitões a mais no total de nascidos para o tratamento com hormônio, não havendo diferença significativa quando considerado o fator \"flushing\". A analise econômica feita pelas observações obtidas no presente estudo, considerando taxa de aproveitamento do parto e total de nascidos, destaca benefício financeiro apresentado através do valor presente líquido (VPL) para a combinação hormonal de $1.862,75, enquanto que o tratamento que utilizou indução somente com o macho, revelou um VPL negativo de $2.845,55. Conclui-se que houve efeito positivo das gonadotrofinas (eCG e LH) na indução e sincronização do estro à puberdade o qual pode ser associado, na menor dispersão da ciclicidade das marrãs até o quarto estro, no menor número de fêmeas descartadas até o primeiro parto e na vantagem econômica com base na taxa de aproveitamento das fêmeas até o parto e total de leitões nascidos. / The objective of the study was to investigate in a biotecnical program applied in a commercial breeding unit, the effects of the gonadotrophins (eCG and LH) on puberty estrus inductions and synchronization in gilts followed by their ciclicity. The investigation included the use of flushing on preceding oestrus cycle to first artificial insemination (IA). The traits analised were: conception rate, farrowing rate, the percentage of animals that started the experimental period and stayied to first farrowing, and litter size. The experiment used 119 hybrid gilts on 153 days of age. The experimental design was entirely random in a factorial arrangement 2x2. One factor corresponding to the gonadotrophin (H) application or the male (M) induction only. The hormonal treatment utilized a combination of 600 UI of eCG (Novormon®, Syntex S. A., Argentina) and 72 hours lates, 2,5 mg of LH (Lutropin®, Vetrepharm Canadá Inc., Canadá). The other factor corresponds to the flushing application in a restriction - ad libitum regimen (R) or a flushing application based on a lactation diet offered in an ad libitum regimen in the breeding unit (B). This factor was applied in the oestrus cycle that preceded first artificial insemination, occurring at 220 days of age. Our results identified,18,33% hormonal treatment of females showing apparent estrus compared to 5% in treatment that used only the male induction. From 18 to 30 days after induction, considering the estrus interval variation about 18-25 days, the hormonal treatment showed significant percentage more than the male induction (48,33% vs. 16,94%, respectively). In the following intervals the differences weren\'t statiscally significant. The cumulative percentuals until 30 days and 90 days periods, the values were 71,67% vs. 69,49% and 91,67% vs. 94,92%, respectively, for hormonal treatment and male induction. There wasn\'t significant interation for reproductive traits. There wasn\'t significant difference on the conception rate, when you considered the factors separately, hormonal treatment (H) and male (M) induction (96,23% vs. 94,00%, respectively) and flushing (R) and the regime adapted in the breeding unit (B) (95,92% vs. 94,44%, respectively). While the farrowing rate hasn\'t showed significance, there was umerical differences in favour of hormonal treatment against male induction (92,45 %vs 80,00%, respectively) (P=0,0653). Considering the period since induction (153 days of age) until parturition, the percentual of hormonal treatment group was superior compared to the male induction group (81,67% vs. 67,80%, respectively) (P=0,0815). However, the percentual was some flushing (R) and the regimen assumed in the breeding unit (G) (70,00% vs. 67,80%, respectively). There wasn\'t significant difference related to total born, born alive, stillborn and mummified fetus on both factors. It is important to emphasize the numerical differences of 0,75 total born piglets in favor of the hormonal groups. There wasn\'t any difference on the flushing factor. The economical analyses, considering the period since induction until parturition and total born piglets suggested a financial benify to the hormonal treatment compared to the male induction.

O estudo objetivou avaliar num programa biotécnico aplicado a uma granja comercial, os feitos combinados, emprego de gonadotrofinas e aplicação do \"flushing\". As gonadotrofinas empregadas na indução e sincronização do estro à puberdade em marrãs, aliado a ciclicidade até o 4º estro, correspondeu à aplicação ou não da combinação hormonal, 600 UI de eCG (Novormon®, Syntex S. A., Argentina) e após 72 horas, 2,5mg de LH porcino (Lutropin®, Vetrepharm Canadá Inc., Canadá). O \"flushing\" alimentar ou esquema alimentar adotado na granja, foi empregado no ciclo estral que antecedeu a primeira inseminação artificial, ocorrida aos 220 dias de idade das fêmeas. Os parâmetros analisados foram: taxa de concepção (TC), taxa de parto (TP), taxa de aproveitamento (TA) e tamanho de leitegada (TL), sendo esse último representado pelo número de leitões nascidos totais (NT), nascidos vivos (NV), natimortos (NM) e mumificados (MM). Foram utilizadas 119 leitoas híbridas, num delineamento experimental inteiramente casualizado em arranjo fatorial 2x2. O tratamento com hormônio mostrou percentual significativamente maior de fêmeas que manifestaram estro até o 5° dia de início da indução, em comparação com o tratamento somente com o estímulo do macho, 18,33% vs. 5,08%, respectivamente (P=0,0249). No intervalo seguinte de 18 - 30 dias após a indução, considerando variação do ciclo estral de 18 a 25 dias, a combinação hormonal revelou percentual significativamente maior comparado com o estímulo natural do macho, 48,33% vs. 16,94%, respectivamente (P=0,0003). Nos intervalos subseqüentes as diferenças não foram significativas. Não foi detectada interação significativa para os efeitos combinados gonadotrofinas e \"flushing\". Na análise das características em separado, não houve diferença significativa na taxa de concepção para os tratamentos combinação hormonal e estímulo do macho (96,23% vs. 94,00%, respectivamente), e tratamentos com \"flushing\" e esquema alimentar da granja (95,92% vs. 94,44%, respectivamente). Na taxa de parto, os percentuais embora não tenham revelado significância, mostraram diferenças numéricas, sendo o maior valor numérico representado pelo tratamento com hormônio em comparação com o macho, 92,45% vs. 80,00% respectivamente (P=0,0653). Quanto à taxa de aproveitamento das fêmeas, considerando a indução aos 153 dias de idade das marrãs até o parto, da mesma maneira evidenciou-se diferença numérica mostrando o tratamento com hormônio percentual maior em comparação com macho, 81,67% vs. 67,80%, respectivamente (P=0,0815). Não houve diferença significativa nos tratamentos relacionados à aplicação do \"flushing\" e esquema alimentar da granja, 70,00% vs. 79,66%, respectivamente. Não foi evidenciada diferença significativa quanto ao total de nascidos, nascidos vivos, natimortos e mumificados, para os dois fatores. Destacou-se diferença numérica de 0,75 leitões a mais no total de nascidos para o tratamento com hormônio, não havendo diferença significativa quando considerado o fator \"flushing\". A analise econômica feita pelas observações obtidas no presente estudo, considerando taxa de aproveitamento do parto e total de nascidos, destaca benefício financeiro apresentado através do valor presente líquido (VPL) para a combinação hormonal de $1.862,75, enquanto que o tratamento que utilizou indução somente com o macho, revelou um VPL negativo de $2.845,55. Conclui-se que houve efeito positivo das gonadotrofinas (eCG e LH) na indução e sincronização do estro à puberdade o qual pode ser associado, na menor dispersão da ciclicidade das marrãs até o quarto estro, no menor número de fêmeas descartadas até o primeiro parto e na vantagem econômica com base na taxa de aproveitamento das fêmeas até o parto e total de leitões nascidos. / The objective of the study was to investigate in a biotecnical program applied in a commercial breeding unit, the effects of the gonadotrophins (eCG and LH) on puberty estrus inductions and synchronization in gilts followed by their ciclicity. The investigation included the use of flushing on preceding oestrus cycle to first artificial insemination (IA). The traits analised were: conception rate, farrowing rate, the percentage of animals that started the experimental period and stayied to first farrowing, and litter size. The experiment used 119 hybrid gilts on 153 days of age. The experimental design was entirely random in a factorial arrangement 2x2. One factor corresponding to the gonadotrophin (H) application or the male (M) induction only. The hormonal treatment utilized a combination of 600 UI of eCG (Novormon®, Syntex S. A., Argentina) and 72 hours lates, 2,5 mg of LH (Lutropin®, Vetrepharm Canadá Inc., Canadá). The other factor corresponds to the flushing application in a restriction - ad libitum regimen (R) or a flushing application based on a lactation diet offered in an ad libitum regimen in the breeding unit (B). This factor was applied in the oestrus cycle that preceded first artificial insemination, occurring at 220 days of age. Our results identified,18,33% hormonal treatment of females showing apparent estrus compared to 5% in treatment that used only the male induction. From 18 to 30 days after induction, considering the estrus interval variation about 18-25 days, the hormonal treatment showed significant percentage more than the male induction (48,33% vs. 16,94%, respectively). In the following intervals the differences weren\'t statiscally significant. The cumulative percentuals until 30 days and 90 days periods, the values were 71,67% vs. 69,49% and 91,67% vs. 94,92%, respectively, for hormonal treatment and male induction. There wasn\'t significant interation for reproductive traits. There wasn\'t significant difference on the conception rate, when you considered the factors separately, hormonal treatment (H) and male (M) induction (96,23% vs. 94,00%, respectively) and flushing (R) and the regime adapted in the breeding unit (B) (95,92% vs. 94,44%, respectively). While the farrowing rate hasn\'t showed significance, there was umerical differences in favour of hormonal treatment against male induction (92,45 %vs 80,00%, respectively) (P=0,0653). Considering the period since induction (153 days of age) until parturition, the percentual of hormonal treatment group was superior compared to the male induction group (81,67% vs. 67,80%, respectively) (P=0,0815). However, the percentual was some flushing (R) and the regimen assumed in the breeding unit (G) (70,00% vs. 67,80%, respectively). There wasn\'t significant difference related to total born, born alive, stillborn and mummified fetus on both factors. It is important to emphasize the numerical differences of 0,75 total born piglets in favor of the hormonal groups. There wasn\'t any difference on the flushing factor. The economical analyses, considering the period since induction until parturition and total born piglets suggested a financial benify to the hormonal treatment compared to the male induction.

Gonadotropin-releasing hormone (GnRH) and its superactive analogues are currently being used in the treatment of a number of endocrine disorders, such as endometriosis, precocious puberty, infertility and prostatic cancer. Selection of these analogues for clinical use have been previously based on their activities in animal models. This thesis has therefore investigated the binding characteristics of the human GnRH receptor, in comparison to those of the rat receptor, as well as the activities of a number of GnRH analogues for stimulating luteinising hormone (LH) and follicle stimulating hormone (FSH) secretion from cultured human pituitary cells. The establishment of a human pituitary bioassay system has further made possible the investigation of the direct regulatory roles of GnRH and other neuropeptides in man. To date, such studies in man have been performed in vivo and are thus complicated by the simultaneous interactions of numerous modulators.