BACKGROUND:Seminal fluid plays an important role in successful fertilization, but knowledge of the full suite of proteins transferred from males to females during copulation is incomplete. The list of ejaculated proteins remains particularly scant in one of the best-studied mammalian systems, the house mouse (Mus domesticus), where artificial ejaculation techniques have proven inadequate. Here we investigate an alternative method for identifying ejaculated proteins, by isotopically labeling females with 15N and then mating them to unlabeled, vasectomized males. Proteins were then isolated from mated females and identified using mass spectrometry. In addition to gaining insights into possible functions and fates of ejaculated proteins, our study serves as proof of concept that isotopic labeling is a powerful means to study reproductive proteins.RESULTS:We identified 69 male-derived proteins from the female reproductive tract following copulation. More than a third of all spectra detected mapped to just seven genes known to be structurally important in the formation of the copulatory plug, a hard coagulum that forms shortly after mating. Seminal fluid is significantly enriched for proteins that function in protection from oxidative stress and endopeptidase inhibition. Females, on the other hand, produce endopeptidases in response to mating. The 69 ejaculated proteins evolve significantly more rapidly than other proteins that we previously identified directly from dissection of the male reproductive tract.CONCLUSION:Our study attempts to comprehensively identify the proteins transferred from males to females during mating, expanding the application of isotopic labeling to mammalian reproductive genomics. This technique opens the way to the targeted monitoring of the fate of ejaculated proteins as they incubate in the female reproductive tract.

The objective of this research was to clarify the basis of subfertility in Delaware roosters. It was anticipated that a sensitive method would be needed to compare seminal plasma protein composition between subfertile and fertile roosters. Consequently, the applicability of two-dimensional electrophoresis was tested as a tool for the analysis of chicken seminal plasma proteins. Two-dimensional electrophoresis resolved 95 ± 4.4 derivative polypeptides from seminal plasma proteins of fertile roosters, whereas one-dimensional electrophoresis resolved only 23 ± 0.4. Thus, two-dimensional electrophoresis was found to be a useful tool for seminal plasma protein analysis. Seminal plasma composition was compared between subfertile Delaware and fertile roosters. Seminal plasma from subfertile roosters was characterized by an imbalance of proteins, electrolytes, and amino acids (P<0.05). Neither type of seminal plasma contained proteolytic activity. No difference (P>0.05) was observed in seminal plasma osmolality. Differences in seminal plasma composition were attributed to a dysfunction of the excurrent ducts of the testis. This realization lead to experiments designed to modulate subfertility. Hemicastration exacerbated (P<0.001) subfertility, whereas supplementation of spermatozoa with seminal plasma proteins from fertile roosters ameliorated (P<0.001) subfertility. Addition of seminal plasma proteins from subfertile roosters to spermatozoa from fertile roosters had no effect (P>0.05) on fertility. Therefore, subfertility was attributed to protein deficiency in seminal plasma rather than the presence of some agent that induces subfertility. The study of subfertile Delaware roosters has helped establish a link between seminal plasma proteins and fertility in the domestic fowl. / Graduation date: 1992

published_or_final_version / Anatomy / Master / Master of Philosophy

Thesis (Ing. Zootecnista)--Universidad Católica Boliviana San Pablo, Unidad Académica Campesina Tiahuanaco, Carrera Ingeniería Zootécnica, 2002. / Reproduced from copy at BYU's Benson Institute. Includes bibliographical references (leaves 87-91).

Thesis (M. Phil.)--University of Hong Kong, 1994. / Includes bibliographical references (leaves 129-157).

Thesis (Ph. D.)--Virginia Polytechnic Institute and State University, 1995. / Vita. Abstract. Includes bibliographical references (leaves 64-68). Also available via the Internet.

No description available.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Em altas latitudes tem-se observado uma variaÃÃo na composiÃÃo bioquÃmica do plasma seminal (PS) de caprinos por influÃncia da estacionalidade. Em baixas latitudes essa variaÃÃo tem sido atribuÃda unicamente ao baixo valor nutricional das dietas na Ãpoca seca. Objetivou-se por meio deste estudo avaliar a ocorrÃncia de variaÃÃo na composiÃÃo bioquÃmica do PS de caprinos SPRD criados intensivamente no Estado do CearÃ, Brasil. O experimento foi conduzido nas instalaÃÃes do Dep. de Zootecnia da Universidade Federal do Cearà â UFC. Foram utilizados 20 bodes SPRD, alimentados segundo o NRC/caprinos (1981). Os ejaculados foram coletados semanalmente em vagina artificial e letroejaculador e em seguida avaliados, mensurados, centrifugados e separados o PS (sobrenadante) das cÃlulas espermÃticas (precipitado), sendo acondicionados em tubos eppendorffs a -18ÂC, durante um ano (Set/2005 a Ago/2006). Para realizaÃÃo das anÃlises as amostra semanais passaram a constituir um pool individual mensal, onde foram avaliadas as concentraÃÃes de Ca, P, Mg, PTt, AC e frutose. Foi observada variaÃÃo significativa (p<0,05) na composiÃÃo bioquÃmica do PS entre as Ãpocas do ano, registrando-se os nÃveis mais elevados dos parÃmetros bioquÃmicos na Ãpoca chuvosa. A variaÃÃo individual foi significativa (p<0,01), sà havendo interaÃÃo significativa, animal/Ãpoca para a frutose (p<0,05). O estudo das correlaÃÃes mostrou associaÃÃes significativas (p<0,0001) entre PTt, AC e frutose, entre os volumes de ejaculado e de PS, com os componentes, e de temperatura, umidade, precipitaÃÃo e ITU, com os parÃmetros bioquÃmicos do PS. Concluindo-se que a frutose à um bom parÃmetro bioquÃmico para o estudo da qualidade seminal, entretanto, mais estudos sÃo necessÃrios para identificar as causas desta variaÃÃo, e compreender melhor o papel da frutose, acido cÃtrico e proteÃnas totais no metabolismo espermÃtico. / Goat seminal plasma (SP) biochemical composition observed to very in high latitudes due to seasonality. In low latitudes this variation has been attributed only to the low nutritional value of diets in the dry period. The aims of this study were to evaluate the occurrence of variation in the SP biochemical composition of SPRD goats breed intensively in the State of CearÃ, Brazil. The experiment was carried out in the facilities of Dep. of Zootecnia of the Federal University of Cearà - UFC. Twenty male SPRD goats were used, they were fed according to NRC/goats (1981). Ejaculated were collected weekly in artificial vagina and eletroejaculador, and evaluated, measure, centrifuged and separated the SP (sobreswing) from the cells spermatic (precipitate), being stored in eppendorff tubes at -18ÂC, during one year (Sep/2005 the Aug/2006). For analyses purpose weekly sample were mixed to constitute a monthly individual pool, from which were evaluated the concentrations of Ca, P, Mg, PTt, CA and fructose. Significant variation was observed (p <0,05) in the biochemical composition of the SP among the periods of the year, registering the highest levels of biochemical parameters in the rainy period. The individual variation was significant (p <0,01), only having significant interaction animal/period to the fructose (p <0,05). The study of correlations showed significant associations (p <0,0001) among PTt, CA and fructose, among the ejaculate volume and of SP with the components, and of temperature, humidity, precipitation and ITU with the biochemical parameters of the SP. It is concluded that fructose is a good biochemical parameter for the study of seminal quality. However, more studies are necessary to identify the causes of this variation, and to better understand the role of the fructose, citric acid and total proteins in the metabolism spermatic.

Made available in DSpace on 2014-06-11T19:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-17Bitstream added on 2014-06-13T18:51:13Z : No. of bitstreams: 1 motheo_tf_me_jabo.pdf: 1290385 bytes, checksum: ce65f89bddd3c29c19638e90cb3193d2 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Atualmente, estudos têm demonstrado a utilização da toxina botulínica do tipo A (TB-A) no tratamento de afecções prostáticas, como a hiperplasia prostática benigna. Ainda, sabe-se que alguns componentes bioquímicos do plasma seminal são relativamente específicos para a regulação da função espermática. O objetivo deste trabalho foi avaliar os possíveis efeitos deletérios ou benéficos da TB-A sobre a fertilidade de cães com HPB. Foram utilizados 18 cães machos, sem raça definida com sinais de HPB. Os animais foram divididos ao acaso em 3 grupos de 6 cães que receberam injeção intraprostática de solução salina de NaCl 0,9% (GC), solução contendo 250U (GI) e 500U de TB-A (GII). Quatro amostras foram colhidas antes da aplicação e 2, 4 e 8 semanas após o tratamento. Foi mensurado o pH e dosadas as concentrações de proteína total, cloretos totais, cálcio, potássio e sódio das amostras de plasma seminal. Ainda, foi realizada eletroforese SDS-Page utilizando géis de poliacrilamida nas concentrações de 12 e 18%. Os resultados foram avaliados por meio de análise de variância (ANOVA) e ao teste de Kruskall-Wallis (p<0,05). Não foram observadas diferenças significativas nos parâmetros bioquímicos avaliados. Outrossim, foram constatadas 31 bandas protéicas, com pesos moleculares variando de 106,2 a 3,9 kDa semelhantemente aos achados descritos na literatura. Destarte, pode-se concluir que os tratamentos com 250U e 500U de TB-A não alteraram os perfis bioquímico e protéico do plasma seminal de cães com HPB e, portanto podem ser considerados uma boa opção para cães destinados à reprodução ou envolvidos em programas de criopreservação de gametas. / Recently, botulinum neurotoxin type A (BoNT-A) application in the lower urinary tract has been extended to prostate disorders such as benign prostatic hyperplasia (BPH). Nevertheless, sperm function is highly dependent on ionic and protein environment. The goal of the present study was to evaluate the effects of BoNTA treatment on seminal plasma biochemical and eletrophoretic profile in dogs with BPH. Eighteen mature male mongrel dogs with BPH, were use in this study. They were randomly divided in three groups, which received intraprostatic injection of 0,9% NaCl normal saline solution (GI), 250U(GII) and 500U (GIII) of BoNT-A. All animals were previously conditioned to semen collection and four semen samples were collected before treatment and at 2, 4 and 8 weeks after injection. The semen was evaluated, and centrifuged to obtain seminal plasma for measurement of pH, total protein concentration (TP), total chlorides (TC), calcium (Ca), potassium (K), and sodium (Na). Additionally, one-dimensional sodium dodecyl sulfatepolyacrilamide gel eletrophoresis (SDS-PAGE) was carried out on 12 and 18% vertical minigel. One way ANOVA and Kruskall-Wallis with Bonferoni as a post hoc test were used to comparison of means (P<0.05). Average pH, TP, TC, Ca, K did not vary significantly at any time point and among each studied group and maintained normal values for the specie. Similarly as described in the literature, the eletrophoresis analysis of the pooled eluded fractions, identified 31 bands, with molecular weights varying from 106.2 to 3.9 kDA, In conclusion, the treatment with 250U and 500U of BoNT-A did not modify the biochemical or protein profiles of the seminal plasma of dogs with BPH.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Um dos principais objetivos de um programa de inseminaÃÃo artificial em caprinos à produzir o maior nÃmero de descendentes de animais geneticamente superiores com a finalidade de melhorar a produÃÃo animal. Para tanto à necessÃrio que se dominem tÃcnicas de diluiÃÃo e de armazenamento de sÃmen nesta espÃcie. Neste trabalho foram testados trÃs diferentes diluidores, citrato-gema (CG), TRIS-gema (TG) e Ãgua de coco industrializada (ACI) e dois grupos de animais, com alto nÃvel inicial de frutose (grupo I) e com baixo nÃvel inicial de frutose (grupo II) no plasma seminal. O sÃmen foi armazenado por atà 48h a 5oC e avaliado nos tempos 0 (fresco), 2, 24 e 48h, onde uma amostra do sÃmen conservado foi submetida ao teste de termo-resistÃncia (TTR) a 38oC por 2h. Durante a incubaÃÃo foram verificados os parÃmetros de motilidade (MOT) e vigor (VIG), nos tempos 5, 60 e 120 minutos e ao final foi calculada a taxa de degradaÃÃo da motilidade (TDM). EsfregaÃos de sÃmen foram realizados para verificaÃÃo das possÃveis alteraÃÃes morfolÃgicas. Houve uma queda significativa nos parÃmetros analisados no decorrer do tempo de armazenamento, independente do diluidor utilizado (P<0,05). Contudo, o diluidor ACI apresentou resultados inferiores aos dos demais. O efeito do grupo foi observado apenas no ACI, mostrando que este diluidor nÃo conseguiu suprir as necessidades energÃticas dos espermatozÃides cuja concentraÃÃo inicial de frutose no plasma estava baixa. Os diluidores CG e TG comportaram-se de forma semelhante nos dois grupos. Foi encontrada diferenÃa significativa na atividade da fosfolipase A2 (PLA2) entre os grupos (P<0,05), sendo que uma atividade superior foi observada no grupo II. Concluiu-se que, independente do diluidor utilizado, os parÃmetros seminais tendem a cair conforme o tempo de conservaÃÃo à prolongado. AlÃm disso, a concentraÃÃo inicial de frutose nÃo interferiu na qualidade espermÃtica quando os diluidores CG e TG foram utilizados, todavia, quando os nÃveis deste componente no plasma seminal foram superiores a 740 mg/dL ocorreu um aumento na sobrevida dos espermatozÃides conservados a 5 oC, quando se utilizou ACI. Sugere-se que o baixo desempenho espermÃtico tambÃm possa ser devido a uma maior atividade de PLA2, contudo mais estudos, contendo um maior nÃmero de animais, sÃo necessÃrios para elucidar esta hipÃtese / One of the main objectives of the artificial insemination program in goats is to produce the highest number of descendants of genetically superior animals with the purpose to improve the animal production. Thus, it is necessary to dominate correct techniques of dilution and semen storage in this specie. In this work three different extenders had been tested: citrate-egg yolk (CE), TRIS-egg yolk (TE) and industrialized coconut water (ICW) and two groups of animals: with high initial level of fructose (group I) and with low initial level of fructose (group II) in the seminal plasma. The semen was stored until 48h at 5oC and evaluated in times 0 (fresh), 2, 24 and 48h, where a sample of the conserved semen was submitted to the term-resistance test (TTR) that consists of incubate the sample at 38oC for 2h. During the incubation it had been verified the parameters of Motility (MOT) and vigor (VIG), in times 5, 60 and 120 minutes and to the end was calculated the tax of degradation of the motility (TDM) and semen slides had been carried through for verification of the possible morphologic alterations. It had a significant fall in the parameters analyzed in elapsing of the time of storage, independent of the used extender (P<0,05). However, extender ICW presented inferior resulted than the others. The effect of the group was observed only in the ICW,showing that this extender did not supply the energy necessities of the spermatozoa whose initial concentration of fructose in the plasma was low. CE and TE worked similarly in the two groups. Significant difference in the activity of phospholipase A2 between the groups was found (P<05), in way that a bigger activity was observed in group II. In conclusion, independent of the used extender, the seminal parameters tend to fall as draw out the conservation time. Moreover, the initial concentration of fructose did not intervene with the sperm quality when extenders CE and TE had been used, however, when the levels of this component in the seminal plasma had been superior to 740mg/dL, occurred an increase in lifetime of the spermatozoa conserved at 5oC, when ICW was used. It suggests that sperm low action also can be due to a higher activity of PLA2, however more studies are necessary, with a large number of animals to elucidate this hypothesis