Orientador: Wilma de Grava Kempinas / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-10T14:12:01Z (GMT). No. of bitstreams: 1 Pontes_DaviAbeid_M.pdf: 1952879 bytes, checksum: ba9688c1750c742fa3f40d3d422ed3eb (MD5) Previous issue date: 2008 / Resumo: O Diabetes mellitus geralmente se associa a algum tipo de disfunção sexual, provocando infertilidade, tanto em seres humanos, quanto em animais experimentais. Em trabalho anterior realizado em nosso laboratório, ratos machos que tiveram hiperglicemia induzida quimicamente pela administração de streptozotocin apresentaram diminuição da fertilidade através de múltiplos parâmetros analisados. O presente estudo teve como objetivo investigar os mecanismos envolvidos e o papel da testosterona no processo. Para tanto, ratos machos foram divididos em 3 grupos experimentais: normoglicêmico (controle), hiperglicêmico (streptozotocin), e hiperglicêmico com reposição hormonal (streptozotocin+testosterona) e foram avaliados parâmetros reprodutivos e espermáticos, níveis hormonais, a contratilidade do ducto deferente isolado in vitro, comportamento sexual e o número de espermatozóides ejaculados no útero. O ducto deferente de animais diabéticos apresentou um quadro de hipersensibilidade à metoxamina, um agonista sintético de adrenoceptores _1. Estes mesmos animais apresentaram os seguintes resultados: alteração de comportamento sexual e ausência de espermatozóides ejaculados, redução dos níveis plasmáticos de testosterona, perda de peso corpóreo e de órgãos como epidídimo, ducto deferente, vesículas seminais e próstata ventral, perda de células germinativas na luz e desorganização epitelial aparente em túbulos seminíferos, além da aceleração do tempo de trânsito dos espermatozóides pelo epidídimo. Os dados apresentados neste trabalho indicam que os mecanismos responsáveis pela perda de fertilidade natural de ratos diabéticos envolvem comprometimento do processo espermatogênico, assim como desregulação do eixo reprodutivo masculino, juntamente com evidências para problemas no processo de maturação espermática, tendo como fator complicante o prejuízo da função ejaculatória, dependente da contratilidade da musculatura lisa dos ductos deferentes. A reposição de andrógeno não foi totalmente capaz de reverter os danos causados pelo diabetes no sistema reprodutivo masculino de ratos adultos / Abstract: Diabetes mellitus is usually related with some kind of sexual dysfunction, promoting infertility in humans as well as in experimental models. In a prior work from our laboratory, male rats, which had a diabetic-induced state of hyperglycemia caused by streptozotocin administration, demonstrated reduced fertility through several parameters analyzed. The present study aimed at investigating the mechanisms involved and the role of testosterone in the process. Male rats were randomly allocated in 3 experimental groups: control, hyperglycemic (streptozotocin), and hyperglycemic with hormone replacement (streptozotocin+testosterone) and the following parameters were analyzed: reproductive and spermatic parameters, hormone levels, sexual behavior, contractility of vas deferens in vitro, sexual behavior parameters and the number of sperm ejaculated in utero. The vas deferens of diabetic animals was hypersensitive to methoxamine, a synthetic agonist of _1 adrenoceptors. The same animals showed the following results: alterations in sexual behavior and lack of sperm ejaculated, reduction in plasma testosterone levels, decreased body weight and epididymis, seminal vesicles, ventral prostate and vas deferens weights, loss of germ cells in the lumen and apparent epithelial disarrange in seminiferous tubules, and accelerated sperm transit time in the epididymis. The data presented herein indicate that the mechanisms underlying the reduced fertility through natural mating observed in diabetic rats involve impairment of the spermatogenic process, as well as a dysregulation of the male reproductive axis, together with evidence for problems in the sperm maturation process, which has as a complicant factor the impairment of the ejaculatory function, dependent on the vas deferens smooth muscle contractility. Androgen replacement was not totally capable of reversing the damage caused by diabetes on the male reproductive system of adult rats / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

Orientador: Rejane Maira Goes / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-05T20:48:06Z (GMT). No. of bitstreams: 1 Pinto_MariaEtelvina_M.pdf: 13780138 bytes, checksum: 687200b70b6f4ab0abe1ba1bd2d7243a (MD5) Previous issue date: 2005 / Resumo: Embora seja bem conhecido que o estrógeno é essencial para o adequado desenvolvimento embrionário e maturação pós-natal do testículo, seu papel específico para a fisiologia testicular e em especial para a espermatogênese ainda permanece pouco compreendido. O objetivo do presente estudo foi determinar o impacto da administração subcutânea de uma dose única (35 mg/kg de peso corporal) de benzoato de estradiol e do anti-estrógeno Tamoxifeno sobre o testículo de ratos jovens (5 semanas de idade). Os efeitos do desequilíbrio hormonal passageiro foram avaliados uma (curto prazo) e sete semanas (médio prazo) após os referidos tratamentos com o uso de análises de rotina em microscopia de luz e microscopia eletrônica de transmissão, associadas a análises morfométricas e estereológicas e também pelo método de TUNEL para a detecção de células apoptóticas. O estrógeno afetou mais drasticamente a estrutura testicular e a espermatogênese quando comparado com o Tamoxifeno. Em curto prazo, a exposição à alta dose de estrógeno reduziu em 50% o peso testicular, diminuiu o diâmetro dos túbulos seminíferos e a população de células de Sertoli, induziu um aumento na apoptose das espermátides alongadas acarretando seu desaparecimento. Embora o índice gônadosomático tenha se recuperado sete semanas após a injeção de estrógeno, a taxa de apoptose das células germinativas ainda permanece 5 vezes mais alta e o número de espermátides alongadas é muito inferior ao encontrado nos animais controle. Em curto prazo, a exposição ao Tamoxifeno foi menos prejudicial para o testículo de ratos jovens em comparação com o estrógeno, mas, em médio prazo, resultou em alguns efeitos semelhantes tais como redução do diâmetro dos túbulos seminíferos, incidência elevada de células germinativas apoptóticas, e atrofia de células de Leydig. A partir dos resultados obtidos concluímos, que tanto a exposição passageira ao estrógeno como ao Tamoxifeno interfere negativamente na dinâmica testicular de ratos jovens, levando a danos na espermatogênese que persistem nos animais adultos com uma provável redução da fertilidade / Abstract: Although it is a consensus that estrogen is essential for normal embryonic development and postnatal maturation of the testis, its specific role to the testicularphysiology and in particular to spermatogenesis, is still little understood The aim of the present research was to determine the impact of a single high dose (35mg1kg of body weight) of estrogen and anti-estrogen Tamoxifen, administered subcutaneously in 5-weekold rats, on the testis structure and spermatogenesis. The effects ofthese transient hormonal disruptions were evaluated one (short-term) and seven weeks (medium-term) after treatments by light and transmission electron microscopies, both associated to morphometric and stereological analysis. Moreover, TUNEL's method was employed in order to detect the apoptotic process on germ cells. The estrogen affected more drastically the testicular structure and the spermatogenesis when compared to Tamoxifen. In short term ana1ysis,the exposure to high doses of estrogen caused a 50% reduction in the testis weight, besides a reduction in the diameter of seminiferous tubules and population of Sertoli cells. This treatment also induced an increase in apoptosis and a total destruction of elongated spermatids. Even though the gonad-somatic-index had been recovered within seven weeks after the estrogen injection, apoptotic germ cell rate still remained tive times higher and the number of elongated spermatids was lower than that found in the control animal group. In short term, the exposure to Tamoxifen was less harrnful to the testis of young rats when compared to the estrogen, but in medium term it resulted in some similar effects such as the reduction of the seminiferous tubule diameter, high occurrence of germ apoptotic cells, and Leydig cell atrophy. Based on these results we conclude that transient exposure to estrogen and Tamoxifen induces aherations in testicular dynamics of young rats with a probable damage to the spermatogenesis of aduh animais / Mestrado / Biologia Celular / Mestre em Biologia Celular e Estrutural

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Spematic cord torsion is one of the more common surgical urological emergencies in childhood. An experimental study has been conducted to investigate acute changes in testicular concentrations of ATP and glucose following one-hour unilateral 720 testicular torsion. Sixty prepubertal male Wistar rats were distributed into two groups: Group 2(Ischemia) and Group 1(Sham operated). Each group(n=30) was divided into 5 subgroups (n=6). All animals were subjected to right testicular torsion of 720Â. The testicular torsion of animals from Group 1 was immediately corrected. Group 2 animals had their testicular torsion lasting one hour followed by detorsion. Testicles were collected and arterial blood samples taken from abdominal aorta of each animal subgroup at the and of ischemic period or one hour after sham operation and 0.5, 1,6 and 24 hours later. ATP and glucose concentrations were significantly decreased (p<0,05) in ischemia group as compared to sham group at the end of ischemic period. There was no difference in glycemia when comparing ischemic versus sham-operated rats. Glucose concentrations were significantly increase (p<0,05) in ipsilateral testis of sham-operated rats No significant change in ATP and glucose concentrations were found in the left testicle in comparasion to the ones measured in the rigth testicle of the same animal. The decrease in ATP and glucose concentrations may be explained by the reduction in blood flow to the torted testis imposed by torsion. The absence of significant contralateral alterations in the concentrations of ATP and glucose could be related to the limited time of isquemia imposed to the right testis. It is concluded that unilateral one-hour testicular torsion causes a decrease in ipsilateral testicular perfusion resulting in decresead testicular concentrations of glucose and ATP in the ipstelial (right) testicles and no alterations in the concentrations of glucose and ATP in the contraleral testicles. Simple testicle manipulation (Sham Operation) causes a signficant increase in glucose concentrations in the ipsilateral(right)testis. / A torÃÃo do cordÃo espermÃtico à a emergÃncia geniturinÃria mais comum na idade pediÃtrica. Foram estudadas as alteraÃÃes metabÃlicas agudas nas concentraÃÃes testiculares de ATP e glicose apÃs uma hora de isquemia induzida por torÃÃo do cordÃo espermÃtico. Sessenta ratos wistar machos prÃ-pÃberes, foram distribuÃdos em 2 grupos de 30 animais cada e cada grupo em 5 subgrupos de 6 ratos. Os ratos do grupo 1 foram submetidos à torÃÃo seguida por distorÃÃo imediata (cirurgia simulada) do testÃculo direito. Os animais do grupo 2 foram submetidos à torÃÃo do testÃculo direito (720o) durante uma hora. Os animais dos diferentes subgrupos foram sacrificados 30 minutos, 1 hora, 6 horas e 24 horas apÃs a distorÃÃo ou realizaÃÃo da cirurgia simulada para a remoÃÃo dos testÃculos e coleta de amostra de sangue arterial. As alteraÃÃes das concentraÃÃes de glicose e ATP foram determinadas por mÃtodos enzimÃticos. Ocorreu queda significante (p<0,05) da concentraÃÃo de glicose e ATP no tempo 0 h, nos testiculos isquemiados. NÃo ocorreram alteraÃÃes na glicemia nem nas concentraÃÃes de glicose e ATP nos testÃculos contralaterais. Os animais submetidos à cirurgia simulada apresentaram aumento significante (p< 0,05) da concentraÃÃo de glicose no testÃculo ipsilateral quando comparada à concentraÃÃo de glicose no testÃculo esquerdo dos mesmos animais nos tempos 0 h e 24h. A diminuiÃÃo do fluxo sangÃÃneo em decorrÃncia da torÃÃo explica a reduÃÃo das concentraÃÃes de ATP e glicose nos testÃculos isquemiados. A ausÃncia de alteraÃÃes nas concentraÃÃes de ATP e glicose nos testÃculos contralaterais (esquerdos) poderia ser uma decorrÃncia do reduzido tempo de isquemia imposto ao testÃculo ipsilateral (direito). Concluiu-se que a torÃÃo testicular unilateral de 720o, por uma hora, induz uma reduÃÃo significante (p < 0,05) das concentraÃÃes testiculares de ATP e de glicose no testÃculo isquemiado sem alteraÃÃes correspondentes da glicemia. A simples manipulaÃÃo do testÃculo direito (cirurgia simulada) induz um aumento da concentraÃÃo de glicose testicular.

Melatonin is a product of the pineal gland and is postulated to play an antigonadotropic role in the reproductive system of mammals. The reproductive system of non-seasonally breeding mammals is believed to be not as responsive to melatonin treatment as that of seasonally breeding mammals. Recently, there has been increasing support from in vivo and in vitro studies, for the hypothesis that melatonin has negative effects on sperm physiology, especially on sperm motility. High and/or low seminal concentrations of melatonin have been associated with abnormalities in human sperm motility and concentration. In this study, I examined the effects of melatonin on the testis, epididymis and sperm physiology, using in vivo and in vitro experiments, in a non-seasonally breeding mammal. Treatment, in vivo, with exogenous melatonin for six weeks did not inhibit testosterone production or spermatogenesis, nor did it affect the mass of the testes and epididymides at dissection, the concentration the morphology of speimatozoa. However, melatonin in vivo had a small, but significant negative effect on sperm motility and sperm motility index. In vitro incubation of spermatozoa Fith melatonin reduced the percentage (%) of forward progressive movement (fpm), increased the % reduction in fpm, reduced the vigor or quality of sperm motility, reduced the sperm motility index, and delayed and/or prolonged the transition of one pattern of sperm motility to the subsequent patterns. Melatonin increased the pH of the culture medium, and the increased pH, and the ethanol utilized as a solvent for melatonin, both negatively affected all the sperm motility parameters that were assessed in my study. The effects of ethanol increased with time, and the effects of pH increased with both time and increasing pH. Melatonin in vitro did not inhibit capacitation and the acrosome reaction, but it delayed the onset and the progression of capacitation and the acrosome reaction. These results suggest that while melatonin did not inhibit spermatogenesis in the Wistar rat, it may influence sperm motility. Therefore, the presence of high concentrations of melatonin in the reproductive fluids may inhibit sperm motility. With further detailed research, melatonin may have a potential use as a contraceptive drug.

Study to determine the seasonal changes in histological and cytological structures in the testis of Huro Salmoides (Lacepede), the large-mouthed black bass.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19243号 / 農博第2140号 / 新制||農||1036(附属図書館) / 学位論文||H27||N4947(農学部図書室) / 32242 / 京都大学大学院農学研究科応用生物科学専攻 / (主査)教授 今井 裕, 教授 祝前 博明, 教授 松井 徹 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Quaternary ammonium compounds (QACs) are antimicrobial disinfectants commonly used in commercial and household settings. While these compounds have been used for decades, reproductive toxicity has not been thoroughly evaluated. Extensive use of QACs results in ubiquitous human exposure to potentially toxic compounds. Reproductive toxicity of two common QACs, alkyl dimethyl benzyl ammonium chloride (ADBAC) and didecyl dimethyl ammonium chloride (DDAC), was investigated to determine gender-specific toxicity with an emphasis on male reproductive function. Breeding pairs of mice exposed for six months to ADBAC+DDAC exhibited decreases in fertility and fecundity, with fewer pregnancies and decreased numbers of pups over a six month period. Females proceeded through significantly fewer estrus cycles, and both ovulation and implantation rates were reduced. Males exhibited declines in both sperm concentration and motility. Male reproductive toxicity was further assessed in a series of in-vitro and in-vivo experiments. ADBAC+DDAC were cytotoxic to testicular Sertoli cells in culture at concentrations greater than or equal to 0.0005%. Changes in blood-testis-barrier integrity (BTB) were observed at 0.01% ADBAC+DDAC using a two-compartment culture system that measures transepithelial electrical resistance (TER). Sertoli cell cytotoxicity correlated with decreased TER at ADBAC+DDAC concentrations above 0.001%. In-vitro fertilization capacity of epididymal sperm was reduced in males given a 10-day rest period following ADBAC+DDAC exposure. Multigenerational changes in sperm parameters and in mRNA expression of enzymes involved with epigenetic modifications were evaluated across three generations. Sperm concentration and motility were reduced in F0 males exposed directly to ADBAC+DDAC. In F1 males, sperm concentration was increased and motility decreased, while there was no change in the F2 progeny. Genes involved in epigenetic modifications were altered in the exposed F0, with upregulation of two histone acetyltransferases (Hat1 and Kat2b) and downregulation of one lysine-specific demethylase (Kdm6b). F1 and F2 generations were not different from controls except for downregulation of the methyltransferase Dnmt1 in F1 progeny. The reproductive toxicity of ADBAC+DDAC identified in these studies, particularly to the male, compels further investigation into the potential effects that these compounds may have on human reproduction. / Ph. D.

Os antígenos cancer-testis (CT) são proteínas imunogênicas expressas em tecido gametogênico e em diferentes tipos de tumor, sendo considerados candidatos promissores para a imunoterapia do câncer. Entretanto, pouco se sabe sobre a função desses antígenos na tumorigênese. Em 2006, identificamos CTSP-1 como um novo antígeno CT, frequentemente expresso em vários tumores. Nesse trabalho, investigamos a função de CTSP-1 por meio da identificação de proteínas expressas em tumores de próstata e que são capazes de interagir fisicamente com esse antígeno. Demonstramos que CTSP-1 interage com a proteína CTCF em ensaios de duplo-híbrido em leveduras, pulldown e de co-localização e, em seguida, analisamos o impacto da superexpressão de CTSP-1 no controle da expressão de genes CT mediada por CTCF e na progressão do ciclo celular. Utilizando o CT NY-ESO-1 como modelo, demonstramos que a superexpressão de CTSP-1 não altera os níveis endógenos de NY-ESO-1 na linhagem celular tumoral H1299. Por outro lado, observamos que a superexpressão de CTSP-1 48h após as transfecções em H1299 induz um bloqueio do ciclo em G0/G1, reduzindo a capacidade clonogênica dessas células por um mecanismo dependente dos níveis de expressão de CTSP-1. Resultados semelhantes não foram observados em ensaios com clones superexpressando CTSP-1 estavelmente, o que sugere que eles tenham se originado de células que conseguiram escapar do bloqueio em G0/G1. Resultados preliminares sugerem que a redução da capacidade clonogênica das células H1299 que superexpressam CTSP-1 48h após as tansfecções não está associada à ocorrência de morte por apoptose. / Cancer-testis (CT) antigens are immunogenic proteins expressed in gametogenic tissues and in different histological types of tumors, being considered promising candidates for cancer immunotherapy. However, little is known about their role in tumorigenesis. In 2006, we identified CTSP-1 as a novel CT antigen, frequently expressed in different types of tumors. In this work, we investigated the functional role of CTSP-1 through the identification of proteins expressed in prostate tumors and that physically interact with this tumor antigen. We demonstrate that CTSP-1 interacts with the CTCF protein using the yeast two-hybrid system, pulldown and co-localization assays and have further analyzed the impact of CTSP-1 overexpression on the expression of CT genes mediated by CTCF and on the cell cycle progression. Using the CT antigen NY-ESO-1 as a model, we showed that the CTSP-1 overexpression does not alter the endogenous levels of NY-ESO-1 in the tumor cell line H1299. On the other hand, we observed that the overexpression of CTSP-1 in H1299 cells 48h after the transfections induces a cell cycle arrest in G0/G1 and reduces the clonogenic capacity of these cells by a mechanism dependent on the CTSP-1 expression levels. Similar results were not observed for cell clones stably overexpressing CTSP-1, suggesting that these clones have arisen from cells that managed to escape cell cycle arrest in G0/G1. Preliminary results suggest that the reduced clonogenic capacity of H1299 cells expressing CTSP-1 and analyzed 48h after the transfections is not associated with cell death by apoptosis.

"April 2002" Includes bibliographical references (leaves 200-249)

"April 2002" / Includes bibliographical references (leaves 200-249) / xv, 249 leaves : ill., plates (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Animal Science, 2003