Foram utilizados 10 sistemas genitais femininos de mocós (Kerodon rupestris) para estabelecer parâmetros de referência relativos à osmolaridade, pH, cálcio, fósforo, uréia, creatinina, glicose e proteínas totais. Os animais foram criados em cativeiro no CEMAS(Centro de Criação de Animais Silvestres), Mossoró – RN. As fêmeas estavam da metade (30-45 dias) para o final da gestação (65-70 dias). As bolsas amnióticas e alantoideanas foram puncionadas individualmente para colheita dos líquidos fetais, que foram centrifugados e analisados posteriormente. Para o líquido amniótico, as concentrações médias em mg/dl foram: glicose = 45,87 ± 22,38; cálcio = 6,31 ± 1,24; fósforo = 1,64 ± 0,72; creatinina = 0,45 ± 0,12; uréia = 34,03 ± 5,96; proteínas totais = 31,24 ± 16,67. Para o líquido alantoideano, as concentrações médias em mg/dl foram: glicose = 59,17 ± 10,85; cálcio = 5,58 ± 0,59; fósforo = 1,27 ± 0,73; creatinina = 0,38 ± 0,38; uréia = 31,49 ± 2,28; proteínas totais = 30,70 ± 18,39. Foram observadas pequenas oscilações entre as concentrações dos parâmetros bioquímicos do fluido amniótico e alantoideano. Estas concentrações são determinadas, provavelmente, pela atividade metabólica materno-fetal. Para os estudos colpocitológicos foram utilizadas 8 fêmeas, sexualmente maduras, mantidas em cativeiro sob condições climáticas tropicais. Foram realizados esfregaços vaginais a cada 72 horas, durante 40 dias. Os animais mostraram ciclo poliestral contínuo, com duração média de 24,25 ± 3,99 dias, não havendo portanto sazonalidade. Durante o proestro houve prevalência de células superficiais anucleadas, intermediárias e parabasais. A fase de estro apresentou índices significativamente mais altos de células superficiais anucleadas e ausência de leucócitos. Alto índice de células intermediárias foi observado no metaestro, com abundância de leucócitos e vacúolos citoplasmáticos (células de Foam). O anestro mostrou índices significativamente mais altos de células parabasais quando comparados com as outras fases do ciclo. Os leucócitos apareceram em número reduzido. / The aim of the present work was to study some reproductives aspects of mocó female (Kerodon rupestris), by assesment of the following parameters: certain biochemical characteristics of fetal fluids and characterization of the estrous cycle by exfoliative colpocytology. In order to establish the levels of the following parameters: osmolarity, pH, calcium, phosphorous, total protein, urea, creatinina, glycose, presents in the amniotic and allantoic fetal fluids, 10 genital systems of mocós were sampled (gestation = 30-45 and 65-70 days). The amniotic and allantoic sacs were punctured and the fetal fluid obtained for posterior analysis. It was observed the following concentrations (mg/ml) in the amniotic fluid: glycose= 45,87 ± 22,38; calcium = 6,31 ± 1,24; phosphorus = 1,64 ± 0,72; creatinine = 0,45 ± 0,12; urea = 34,03 ± 5,96; total protein = 31,24 ± 16,67. In the allantoic fluid the concentrations were: : glycose = 59,17 ± 10,85; calcium = 5,58 ± 0,59; phosphorous = 1,27 ± 0,73; creatine = 0,38 ± 0,38; urea = 31,49 ± 2,28; total protein = 30,70 ± 18,39. Results obtained from the analisys of the composition of fetal fluids showed small oscillations between the amniotic and the allantoic fluids. Their concentrations, probably, are determined by the materno-fetal metabolic activity. Colpocytology studies were realized using 8 adult females kept in captivity, in climatologic conditions of humid tropic. Vaginal smears, prepared by means of swabs, were made each 72 hours, during 40 days. Animals showed a polyestrus continuos cycle (on an average 24,25 ± 3,99 days), so no sazonality was observed. During proestrus phase the exfoliative arrangement showed high frequency of anucleated superficial, intermediate and parabasal cells, respectively. Oestrus phase had elevated rates of superficial cells (both anucleated and nucleated) and absence of leukocytes. In the metestrus phase it was found highest indices of intermediate cells and increase of leukocytes. During the anestrus phase the predominant cells type was parabasal, and leukocytes were reduced.

Fundação de Apoio a Pesquisa e à Inovação Tecnológica do Estado de Sergipe - FAPITEC/SE / Pain can be defined as an unpleasant emotional and sensory experience associated with actual or potential tissue injury. About 70% of the individuals affected by chronic pain are female and the gonadal hormones play a decisive role in terms of sex differences and nociceptive responses. Studies suggest that rodents can recognize pain in co-specifics and expend active behaviors toward it. The main objective of the present study was to evaluate the influence of hormonal changes related to the estrous cycle on the social contact of rats during inflammatory pain and the effect of social contact on the nociceptive responses of female rats with inflammatory pain. Wistar female rats were used at 2 to 3 months of age. The project was composed of two experimental protocols. Experiment I evaluate the behavior of resident rats during exposure to cohabitants with inflammatory pain. It was composed of the following groups and experimental animals: Resident rats (RES), Control (CTRL), Formalin social contact (FCS) and saline (SAL). The three groups were repeated in different cycles stages, and three experiments were carried out where the residents are called Resident Estrous (RES / ES), Resident Ovariectomized (RES / OV) and Resident Proestrus (RES / PRO) All rats but ES and PRO residents underwent ovariectomy surgery. Experiment II was composed of the groups: Formalin Social Contact and the Formalin Isolate, which received the administration of formalin (1%) and was submitted to a 20min isolation period. After the 20 min of contact or isolation the nociceptive responses were evaluated for 40 minutes. The parameters used to evaluate the nociceptive responses were the lick time and the number of flinches. All groups were composed of n = 8. The results showed interaction between the phase the cycle versus treatment (p = 0.0013) for contact duration. In the case of the group of OV residents, (P <0.05) and with those residents in proestrus (p <0.001). In the experiment II, no difference was observed between the FCS and FI groups in groups composed of animals that had contact with ovariectomized or estrus residents. The group composed of animals that had contact with the proestrus residents showed analgesia during thirty minutes of the formalin test between 0 and 30 minutes of observation. The results suggest that rats recognize pain component in a cohabitant and direct active behaviors towards it, however the intensity and quality of this contact may vary according to the cycle. Regarding the nociceptive responses, it was observed that the social contact evoked through direct social behaviors was not enough to elicit analgesia, however aggressive behaviors on the part of the resident elicited an analgesia that may be based on the descending system through RVM with opioid character and being induced by stress. / A dor pode ser definida como uma experiência emocional e sensorial desagradável associada a uma lesão tecidual real ou potencial. Cerca de 70% dos indivíduos que são acometidos por dores crônicas são do sexo feminino e os hormônios gonadais tem função determinante no que tange o diformismo e as respostas nociceptivas. Estudos sugerem que roedores são capazes de reconhecer o componente álgico em co-específicos e despender comportamentos ativos em direção ao mesmo. O objetivo principal do presente trabalho foi avaliar a influência das variações hormonais relativas ao ciclo estral no contato social de ratas durante a dor inflamatória e o efeito do contato social nas respostas nociceptivas de ratas com dor inflamatória induzida por formalina. Foram utilizadas 120 ratas Wistar com 2 a 3 meses de idade. O projeto foi dividido em dois protocolos experimentais. O experimento I teve como objetivo a avaliação do comportamento de ratas residentes durante a exposição a coabitantes com dor inflamatória e foi composto pelos seguintes grupos e animais experimentais: Ratas residentes que foram submetidas aos animais controle (CTRL), formalina contato social (FCS) ou salina (SAL). Essa divisão dos grupos se repetiu nas diferentes fases do ciclo, das ratas residentes que foram denominadas: Residente Estro (ES), Residente Ovariectomizada (OV) e Residente Proestro (PRO). Excetuando as residentes estro e proestro todas as outras ratas foram submetidas a cirurgia de ovariectomia. O experimento II foi composto pelos os grupos: Formalina Contato Social (FCS), e o Formalina Isolado (FI) o qual recebeu a administração da formalina (1%) na pata posterior esquerda e após foi isolada por 20 minutos. Os animais coabitantes dos grupos FCS/ES, FCS/PRO, FCS/OV e FIS tiveram suas respostas nociceptivas avaliadas por 40 minutos. Os parâmetros utilizados para avaliação das respostas nociceptivas foram o tempo de lambida e o número de sacudidas da pata traseira. Todos os grupos foram compostos por n= 8. No que se refere ao Duração do Contato observou-se interação entre fase do ciclo e o tratamento (p= 0,0013). Neste sentido as residentes castradas mostraram duração do contato maiores que as residentes estro (p< 0,05) e as residentes no proestro (p< 0,001). No experimento II não se observou diferença entre os grupos FCS e FI nos grupos compostos por animais que tiveram contato com as residentes castradas e no estro. O grupo composto por animais que haviam tido contato com as residentes proestro mostrou redução no número de sacudidas o período entre 0 e 30 minutos de observação. Os resultados sugerem que ratas são capazes de reconhecer o componente álgico em um coabitante e direcionar comportamentos ativos em pró do mesmo, entretanto a intensidade e a qualidade desse contato podem variar de acordo com o ciclo. No que tange as respostas nociceptivas observou-se que o contato social evocado através de comportamentos sociais diretos não foi suficiente para eliciar analgesia, entretanto comportamentos agressivos por parte da residente eliciaram uma analgesia de longa duração nas coabitantes que pode indicar a ativação do sistema descendente antinociceptivo endógeno através do RVM com caráter opioide e sendo induzida por estresse. / São Cristóvão, SE

O presente estudo teve como objetivo verificar o efeito do uso de gonadotrofinas exógenas na indução do estro à puberdade em marrãs, e \"flushing\" alimentar no ciclo estral que antecede a primeira inseminação artificial na vida útil reprodutiva. O experimento foi realizado na Granja Barreiro (Suinocultura Rosetto) Cerqueira César/SP. Foram utilizadas 96 fêmeas da puberdade ao 1° parto, e 68 fêmeas selecionadas das 96 com ordem de parto do primeiro ao sexto para a avaliação associativa com a vida útil reprodutiva. O delineamento experimental foi inteiramente casualizado em um fatorial 2x2 sendo um fator a combinação hormonal H,( 600 UI de eCG e após 72 horas, 2,5 mg de LH porcino) (H) e o estímulo do macho (M). O fator \"flushing\" alimentar correspondeu ao esquema com restrição (R) e o adotado na granja (G). Os tratamentos foram: HR - hormônio e \"flushing\"; HG - hormônio e esquema alimentar da granja; MR - macho e \"flushing\" e MG - macho e esquema alimentar da granja. As informações foram obtidas a partir do banco de dados existente na propriedade onde o experimento foi realizado. As variáveis análisadas foram: número de estros pós-indução, total de nascidos (TN), nascidos vivos (NV), natimortos (NT), mumificados (MM) e intervalo entre partos (ITP). Cinco dias após a aplicação do tratamento hormonal, 31,37% (H) das fêmeas manifestaram características de estro quando comparado ao tratamento M, onde 11,62% apresentaram estro. O tratamento H mostrou maior sincronismo nas manifestações dos estros, diferente do tratamento M que apresentou maior dispersão ao longo dos ciclos. Não houve diferença estatística nas características de primeiro parto entre os tratamentos hormônio e \"flushing\", tanto quanto nas suas interações (P>0,05). Não houve influência dos tratamentos na produtividade dos animais nas diferentes parições (P>0,05). Portanto, apesar do maior sincronismo apresentado pela combinação hormonal em comparação com o estímulo natural somente pela presença do macho, não houve diferenças quanto à produtividade associativa do primeiro parto e demais partos nas características avaliadas. / The present study objective to verify the effect of the use of exogenous gonadotropins in oestrus induction of puberty in gilts, and \"flushing\" feed the estrous cycle before the first artificial insemination in lifetime reproductive. The experiment was conducted at the Granja Barreiro (Swine Rosetto) - Cerqueira César / SP. A total of 96 females from puberty to the 1st delivery, and 68 of the 96 selected females with birth order from first to sixth for assessing association with lifetime reproductive. The experimental design was completely randomized in a 2x2 factorial being a combination hormonal factor H (600 IU eCG and after 72 hours, 2.5 mg porcine LH) (H) and the stimulation of the male (M). The factor \"flushing\" food matched the schema constraint (R) and adopted on the farm (G). The treatments were: HR - hormone and \"flushing\"; HG - hormone regimen and feed the farm; MR - male and \"flushing\" and MG - male and feed the farm scheme. The information was obtained from the existing database on the property where the experiment was conducted. The variables analyzed were: number of post-estrus induction, total born (TN) live births (NV), stillbirths (NT), mummified (MM) and calving interval (ITP). Five days after application of the treatment, 31,37% (H) of estrus females showed characteristics when compared to treatment M, where 11,62% showed estrous. The H treatment showed greater synchronization of estrus in the demonstrations, unlike the M treatment with the highest dispersion along the cycles. There was no statistical difference in the characteristics of first birth among hormone treatments and \"flushing\" as well as their interactions (P>0.05). No influence of the treatments on the productivity of animals in different parities (P>0.05). Therefore, despite the higher timing presented by combination hormonal stimulation compared with only natural for the presence of males, there were no differences in yield associative first birth and other birth characteristics evaluated.

Para a metabolização de hormônios esteroides sexuais é essencial a participação de enzimas esteroidogênicas. A enzima citocromo P450c17 é responsável pela produção de andrógenos e a enzima citocromo P450 aromatase é responsável pela produção de estrógenos, sendo que, ambas as enzimas necessitam formar um complexo com uma enzima parceira, denominada NADPH citocromo P450 redutase, para realizar a metabolização destes hormônios essenciais para a diferenciação sexual. Objetivou-se imunolocalisar as três enzimas acima citadas no tecido vaginal de fêmeas de roedores Galea spixii. Para tanto, o experimento foi desenvolvido utilizando técnicas de citologia esfoliativa vaginal para definição das fases do ciclo estral, técnicas de microscopia de Luz, Eletrônica de Varredura e testes de imunohistoquímica para detecção das enzimas: citocromo P450c17, citocromo P450 aromatase, e NADPH citocromo P450 redutase. Constatou-se no presente estudo que o ciclo estral das fêmeas de preás silvestres do semiárido apresenta ciclo estral com duração de 15,85 ± 1,4 dias, com a formação e ruptura da membrana de oclusão vaginal durante cada ciclo estral. A vagina apresenta um clitóris hipertrofiado com o óstio uretral que se abre no topo do mesmo, e ausência de um vestíbulo vaginal. O epitélio da mucosa vaginal sofre modificações proliferativas, se espessando e se adelgando de acordo com a respectiva fase do ciclo estral. As enzimas citocromo P450 aromatase e NADPH redutase estão imunolocalizadas durante todo o ciclo estral no epitélio e tecido conjuntivo da vagina de Galea spixii. Por outro lado, a enzima P450c17 está imunolocalizada no epitélio vaginal durante todas as fases do ciclo estral, porém não está presente no estro. Com estes dados pode-se sugerir que ocorre uma metabolização local de hormônios estrógenos e andrógenos no tecido vaginal o que podem estar relacionado com a proliferação celular, variação na vascularização e inervação e ruptura da membrana de oclusão vaginal. / To the metabolism of sex steroid hormones is essential the participation of esteroidogenic enzymes. The enzyme cytochrome P450c17 is responsible for androgen production; and the enzyme aromatase cytochrome P450 is responsible for estrogen production, however, both enzymes require forming a complex with an enzyme partner called NADPH cytochrome P450 reductase to perform the metabolism of these hormones, which are essential for sexual differentiation. This study aimed to immunolocalization of the three enzymes above mentioned in the vaginal tissue from female rodents Galea spixii. Therefore, the experiment was carried out using vaginal cytology techniques for definition of the estrous cycle phases, techniques for light microscopy, scanning electron and immunohistochemical tests for the detection of enzymes: cytochrome P450c17, cytochrome P450 aromatase, and NADPH cytochrome P450 reductase. It was found that the estrus cycle of Galea spixii female lasts 15.85 ± 1.4 days, with the formation and rupture of vaginal closure membrane during each oestrous cycle. The vagina has a hypertrophied clitoris with the urethral orifice that opens on top of it, and it is observed the absence of a vaginal vestibule. The vaginal mucosal epithelium undergoes proliferative changes, with thickening and thinning according to the respective estrous cycle phase. The enzymes cytochrome P450 aromatase and NADPH reductase are immunolocated throughout the oestrous cycle in the epithelium and connective tissue of the vagina from Galea spixii. On the other hand, the enzyme cytochrome P450c17 is immune located in the vaginal epithelium during all stages of the estrous cycle, except at estrous. With these data may be suggested that there is a local estrogens and androgens metabolism in vaginal tissue which may be associated with cell proliferation, vascularization and innervation variation besides the formation and rupture of the vaginal closure membrane.

Orientadores: Claudia Herrera Tambeli, Maria Cecilia F. A. Veiga / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-03T22:56:59Z (GMT). No. of bitstreams: 1 Clemente-Napimoga_JulianaTrindade_M.pdf: 904782 bytes, checksum: 55d51f81e2ba465f145751f287246951 (MD5) Previous issue date: 2004 / Resumo: Este estudo avaliou as diferenças sexuais na resposta nociceptiva induzida pela administração de formalina na articulação temporomandibular (ATM) com ou sem a co-administração do U50,488 (agonista do receptor opióide capa). As fases do ciclo estral das fêmeas foram citologicamente determinadas e apenas aquelas que apresentavam-se na fase diestro ou proestro, e machos foram incluídos. A formalina induziu um comportamento nociceptivo maior nas fêmeas em diestro do que nas fêmeas em proestro ou machos. O U50,488 reduziu significativamente as respostas nociceptivas induzidas pela formalina, e esta redução foi maior nas fêmeas, especialmente nas fêmeas da fase diestro do ciclo estral. A injeção do U50,488 na ATM contralateral não afetou na magnitude do comportamento induzido pela formalina, e o pré-tratamento com o antagonista seletivo do receptor opióide capa nor-binaltorphimine (norBNI) na ATM ipsilateral reduziu os efeitos antinociceptivos do U50,488. Estes resultados demonstram a ação dos receptores opióides capa periféricos na modulação da dor inflamatória. Além disso, considerando que os níveis plasmáticos dos hormônios ovarianos são baixos durante a fase diestro, estes resultados são consistentes com a hipótese de que os hormônios sexuais femininos podem ter uma ação analgésica na redução da dor inflamatória induzida pela formalina, assim como, também ter uma ação anti-analgésica nos efeitos mediados pelos receptores opióides capa / Abstract: This study examined sex differences in nociceptive responses induced by intra-temporomandibular joint (TMJ) formalin with and without co-administration of the ?-opioid receptor agonist U50,488. The estrous phase of females was cytologically determined; only those in either proestrus or diestrus, and males, were included. Formalin elicited significantly greater nociceptive behavior in diestrus females than in either proestrus females or males. U50,488 significantly reduced formalin nociceptive responses, and this reduction was significantly greater in females, especially in the diestrus phase of the estrous cycle. U50,488 injection into the contralateral TMJ failed to affect the magnitude of formalininduced behavior, and preinjection of the selective kappa-opioid receptor antagonist nor-binaltorphimine (nor-BNI) into the ipsilateral TMJ significantly reduced the antinociceptive effect of U50,488. These findings support a role for peripheral kappa-opioid receptors in the modulation of inflammatory pain. Furthermore, since plasma levels of ovarian hormones are lowest during diestrus, these findings are consistent with the suggestion that female sex hormones may play an analgesic role in reducing formalin-induced inflammatory pain, and may also play an anti-analgesic role, at least in ?-mediated effects / Mestrado / Fisiologia Oral / Mestre em Odontologia

Women are twice as likely to suffer from depression as men, yet stress and depression research has relied primarily on the responses of males. Early life stress is hypothesized to influence the development of vulnerability to depression while adult stress exposure can act as a trigger in those predisposed. This relationship is mediated by other environmental factors. Maternal care and the social environment appear to be particularly important for mammals. The purpose of this thesis was twofold: to develop an animal model of depression for use in female rats based on the chronic mild stress (CMS) model previously validated for use in male rodents, and to apply this model in female offspring of mothers exposed to physical restraint in the second half of gestation representing an early life insult. Results indicate that a modified CMS model was able to alter hedonic and physiological responses not present in the original model. Housing condition interacted with CMS in that effects were evident only in singly housed rats. While gestational stress (GS) altered maternal weight and behavioural profiles related to offspring care and anxiety, little to no behavioural effects were noted in juvenile or adult offspring. Applying the modified CMS model to adult female offspring resulted in an anhedonic-like response that recovered rapidly prior to the third week of CMS. Weight in GS female rats was attenuated throughout life beginning post weaning. When taken together, these results demonstrate that stress-based models, previously established in males, must be altered to accommodate the hormonally intact female rat in two ways: first, to eliminate extraneous variables that may interfere with the estrous cycle and mask possible stress effects, and secondly, to consider the appropriateness of individual stressors to induce a stress response in females. While a general lack of effect was noted in response to CMS, this was interpreted as a strong influence of housing and supportive early life experiences in protecting the female rat from the establishment of stress effects related to depression and anxiety. The housing practices employed here may be considered a model of stress-resilience and represents an encouraging avenue of future research.

Made available in DSpace on 2016-01-26T18:55:38Z (GMT). No. of bitstreams: 1 Joao Lawrence Ortigosa.pdf: 235811 bytes, checksum: b02ccf6cce0ff77752f3a8a3d39c1bb7 (MD5) Previous issue date: 2013-06-28 / The aims of this study were to evaluate, in spring and summer (November to March), the plasma progesterone concentration in woolly and hairless ewe sheep created in western São Paulo; evaluate the plasma progesterone concentration as a function of residence time of progestogen (6 , 9 or 12 days) in TAI protocol (fixed-time artificial insemination). Experiment 1: We used 12 sheep cross breeds (6 hairless hair and 6 woolly ewe), in which were made between November and March, eight blood samples (C1 to C8), by jugular venipuncture for later determination of progesterone by radioimmunoassay (RIE). Experiment 2: We used 38 sheep cross breeds randomly divided into three groups: G-6 (n = 13), G-9 (n = 13), and G-12 (n = 12). Initially each sheep received an intravaginal sponge progestogen (D0) that has remained for 6 (G-6), 9 (G-9) or 12 days (G-12). At sponge removal were administered intramuscularly, 0.1315 mg of PGF2a and 300UI eCG. The IATF, laparoscopic, was made from 50 hours after withdrawal of progestin. Thirty days after pregnancy diagnosis was performed by transabdominal ultrasound Exp 1. With the exception of crops 4 (C4) and 8 (C8), all other wooless group P4 concentration showed statistically superior (p <0.05) than the group of lanadas. Exp 2. Upon withdrawal of the progestogen G-12 P4 concentration showed significantly (p <0.05) lower (0.342 ng / mL) to G-6 (1,684 ng / ml) and G-9 (1.762 ng / mL). However there was no difference in pregnancy rates between groups G-6 (76.9%), G-9 (61.5%) and G-12 (91.6%). The hairless ewes showed plasma P4 greater than the woolly sheep in periods. The duration of stay of progestin does not affect the pregnancy rate in ewes. The duration of stay of progestin does not affect the pregnancy rate in sheep, but the sheep in G-12 P4 exhibited lower when compared to G-6 and G-9. / Os objetivos do presente trabalho foram: avaliar, na primavera e verão (novembro a março), a concentração plasmática de progesterona em ovelhas lanadas e deslanadas criadas no oeste paulista; avaliar a concentração plasmática de progesterona em função do tempo de permanência do progestágeno (6, 9 ou 12 dias) em protocolo de IATF (inseminação artificial em tempo fixo). Experimento 1: Foram utilizadas 12 ovelhas mestiças (6 lanadas e 6 deslanadas), nas quais foram feitas, entre novembro e março, oito colheitas de sangue (C1 a C8), por venopunção da jugular., para posterior dosagem de progesterona por radioimunoensaio (RIA). Experimento 2: Foram utilizadas 38 ovelhas mestiças divididas aleatoriamente em três grupos: G-6 (n=13); G-9 (n=13); e G-12 (n=12). Inicialmente cada ovelha recebeu uma esponja intravaginal de progestágeno (D0) que permaneceu por 6 (G-6), 9 (G-9) ou 12 dias (G-12). Na retirada da esponja foram administrados, por via IM, 0,1315mg de PGF2&#945; e 300UI de eCG. A IATF, por via laparoscópica, foi feita a partir de 50 horas após a retirada do progestágeno. Trinta dias após foi realizado diagnóstico de gestação através de ultrassonografia transabdominal. Exp. 1. Com exceção das colheitas 4 (C4) e 8 (C8), em todas as outras o grupo deslanadas apresentou concentração de P4 estatisticamente superior (p<0,05) ao grupo das lanadas. Exp. 2. No momento da retirada do progestágeno o G-12 apresentou concentração de P4 significativamente (p<0,05) menor (0,342 ng/mL) que o G-6 (1,684 ng/mL) e G-9 (1,762 ng/mL). No entanto não houve diferença na taxa de prenhez entre os grupos G-6 (76,9%), G-9 (61,5%) e G-12 (91,6%). As ovelhas DES apresentaram concentração plasmática de P4 maior que as ovelhas L nos períodos avaliados. A duração da permanência do progestágeno não afeta a taxa de prenhez em ovelhas, mas as ovelhas do G-12 exibiram P4 menor quando comparada aos G-6 e G-9.

Made available in DSpace on 2016-07-18T17:53:12Z (GMT). No. of bitstreams: 1 Joao Lawrence Ortigosa.pdf: 235811 bytes, checksum: b02ccf6cce0ff77752f3a8a3d39c1bb7 (MD5) Previous issue date: 2013-06-28 / The aims of this study were to evaluate, in spring and summer (November to March), the plasma progesterone concentration in woolly and hairless ewe sheep created in western São Paulo; evaluate the plasma progesterone concentration as a function of residence time of progestogen (6 , 9 or 12 days) in TAI protocol (fixed-time artificial insemination). Experiment 1: We used 12 sheep cross breeds (6 hairless hair and 6 woolly ewe), in which were made between November and March, eight blood samples (C1 to C8), by jugular venipuncture for later determination of progesterone by radioimmunoassay (RIE). Experiment 2: We used 38 sheep cross breeds randomly divided into three groups: G-6 (n = 13), G-9 (n = 13), and G-12 (n = 12). Initially each sheep received an intravaginal sponge progestogen (D0) that has remained for 6 (G-6), 9 (G-9) or 12 days (G-12). At sponge removal were administered intramuscularly, 0.1315 mg of PGF2a and 300UI eCG. The IATF, laparoscopic, was made from 50 hours after withdrawal of progestin. Thirty days after pregnancy diagnosis was performed by transabdominal ultrasound Exp 1. With the exception of crops 4 (C4) and 8 (C8), all other wooless group P4 concentration showed statistically superior (p <0.05) than the group of lanadas. Exp 2. Upon withdrawal of the progestogen G-12 P4 concentration showed significantly (p <0.05) lower (0.342 ng / mL) to G-6 (1,684 ng / ml) and G-9 (1.762 ng / mL). However there was no difference in pregnancy rates between groups G-6 (76.9%), G-9 (61.5%) and G-12 (91.6%). The hairless ewes showed plasma P4 greater than the woolly sheep in periods. The duration of stay of progestin does not affect the pregnancy rate in ewes. The duration of stay of progestin does not affect the pregnancy rate in sheep, but the sheep in G-12 P4 exhibited lower when compared to G-6 and G-9. / Os objetivos do presente trabalho foram: avaliar, na primavera e verão (novembro a março), a concentração plasmática de progesterona em ovelhas lanadas e deslanadas criadas no oeste paulista; avaliar a concentração plasmática de progesterona em função do tempo de permanência do progestágeno (6, 9 ou 12 dias) em protocolo de IATF (inseminação artificial em tempo fixo). Experimento 1: Foram utilizadas 12 ovelhas mestiças (6 lanadas e 6 deslanadas), nas quais foram feitas, entre novembro e março, oito colheitas de sangue (C1 a C8), por venopunção da jugular., para posterior dosagem de progesterona por radioimunoensaio (RIA). Experimento 2: Foram utilizadas 38 ovelhas mestiças divididas aleatoriamente em três grupos: G-6 (n=13); G-9 (n=13); e G-12 (n=12). Inicialmente cada ovelha recebeu uma esponja intravaginal de progestágeno (D0) que permaneceu por 6 (G-6), 9 (G-9) ou 12 dias (G-12). Na retirada da esponja foram administrados, por via IM, 0,1315mg de PGF2&#945; e 300UI de eCG. A IATF, por via laparoscópica, foi feita a partir de 50 horas após a retirada do progestágeno. Trinta dias após foi realizado diagnóstico de gestação através de ultrassonografia transabdominal. Exp. 1. Com exceção das colheitas 4 (C4) e 8 (C8), em todas as outras o grupo deslanadas apresentou concentração de P4 estatisticamente superior (p<0,05) ao grupo das lanadas. Exp. 2. No momento da retirada do progestágeno o G-12 apresentou concentração de P4 significativamente (p<0,05) menor (0,342 ng/mL) que o G-6 (1,684 ng/mL) e G-9 (1,762 ng/mL). No entanto não houve diferença na taxa de prenhez entre os grupos G-6 (76,9%), G-9 (61,5%) e G-12 (91,6%). As ovelhas DES apresentaram concentração plasmática de P4 maior que as ovelhas L nos períodos avaliados. A duração da permanência do progestágeno não afeta a taxa de prenhez em ovelhas, mas as ovelhas do G-12 exibiram P4 menor quando comparada aos G-6 e G-9.

Ziel der vorliegenden Arbeit war es, die physiologische Zellinfiltration des bovinen Endo-metriums während des Zyklus bei fruchtbaren Rindern (immun-)histologisch zu bestimmen. Darauf aufbauend sollte die Endometritis unter besonderer Berücksichtigung ihres Grades histologisch definiert werden. Des Weiteren erfolgte eine histologische Dokumentation des endometrialen „Gesundheitszustandes“ klinisch gesunder Rinder und es sollte ermittelt wer-den, welche endometrialen Befunde hinsichtlich ihrer Qualität und Quantität mit einer Gravi-dität vereinbar sind und ob eine prognostische Aussage in Bezug auf die Fertilität möglich ist. Zudem wurden unterschiedliche Endometritisformen hinsichtlich Qualität und Quantität der beteiligten Entzündungszellen histologisch und immunhistologisch detailliert charakterisiert. Darüber hinaus erfolgte eine vergleichende Betrachtung der Ergebnisse der histologischen Untersuchung mit denen der Uteruszytologie. Zur Definition des Normalbefundes der endometrialen Infiltration mit freien Zellen wurden nach Ovulationsinduktion von zwölf klinisch-gynäkologisch gesunden Rindern im Östrus (Fol-likelphase) sowie von sieben Kühen zweimal im Laufe eines Zyklus (Zyklustag 6 ± 1, frühe Lutealphase und Zyklustag 14 ± 1, späte Lutealphase) zytologische Präparate und Endo-metriumbioptate entnommen und (immun-)histologisch untersucht. Das Vorkommen von Lymphozyten (T- und B-Lymphozyten), Plasmazellen, neutrophilen und eosinophilen Granu-lozyten, Makrophagen und Mastzellen (Tryptase- und Chymase-positive Mastzellen) wurde quantitativ bestimmt und statistisch untersucht. Mittels dieser Ergebnisse wurden Grenzwerte zwischen der physiologischen endometrialen Infiltration während des Zyklus und dem Vorlie-gen einer Endometritis festgelegt. Weniger als zehn neutrophile Granulozyten sowie weniger als zwölf mononukleäre Zellen (weniger als acht Lymphozyten und weniger als vier Plasma-zellen) pro Gesichtsfeld bei 400facher Vergrößerung im Bereich des Stratum compactum oder des luminalen Epithels mit angrenzenden Anteilen des Stratum compactum werden als Normalbefund angesehen. Um einen Überblick über das Endometrium klinisch gesunder Rinder zu erhalten, wurden Endometriumbioptate von 259 Milchkühen histologisch ausgewertet, wichtige Frucht¬barkeitskennzahlen dokumentiert und die Ergebnisse statistisch untersucht. Von 200 dieser Tiere wurden die Ergebnisse der rektalen Trächtigkeitsuntersuchung herangezogen und die Auswertung der Uteruszytologien in die Untersuchungen integriert. Histopathologisch zeigten sich größtenteils geringgradige Veränderungen. Chronische nicht-eitrige Endometritiden dominierten gegenüber eitrigen Entzündungen. Mit höherem Grad der Endometritis stieg der Besamungsaufwand, während sich der Besamungserfolg verringerte. Lymphfollikel traten vor allem zusammen mit nicht-eitrigen Endometritiden auf. Ein Einfluss der größtenteils gering¬gradigen Endometrose auf die Fruchtbarkeit konnte nicht nachgewiesen werden. Die Angiosklerose stieg mit der Anzahl an Abkalbungen und hatte, insbesondere interkarunkulär, mit höhergradigerer Ausprägung einen negativen Einfluss auf die Fertilität. Karunkuläre Angiosklerosen waren zudem stärker ausgeprägt als inter¬karunkuläre. Zur detaillierten Charakterisierung unterschiedlicher Endometritisformen wurden Endo¬metritiden bei 90 Rindern semiquantitiativ mittels H.E.-Färbung hinsichtlich ihres Grades und Charakters eingeteilt und anschließend die beteiligten Entzündungszellen anhand von Spe-zialverfahren (Immunhistologie, Spezialfärbung) ausgezählt. Vor allem Plasmazellen, neutrophile Granulozyten und T-Lymphozyten weisen im Vergleich zur Zyklusgruppe und mit zunehmendem Grad der Endometritis deutlich erhöhte Zellzahlen auf. Nicht-eitrige Endo-metritiden werden durch die Dominanz von Lymphozyten und insbesondere von Plasmazel-len sowie dem häufigen Vorkommen von Lymphfollikeln zu den chronischen Entzündungen gezählt. Die Entzündung innerhalb der Karunkel entspricht grundlegend in ihrem Charakter und ihrer Ausprägung der des umliegenden interkarunkulären Gewebes. Die endometrialen Zellinfiltrate fanden sich bei dieser Studie sowohl in den verschiedenen Zyklusphasen als auch bei den verschiedenen Endometritiden vor allem in den oberflächli-chen endometrialen Bereichen. Bei allen untersuchten Rindern fiel zudem auf, dass T- Lymphozyten im Endometrium in deutlich höherer Anzahl vorkommen als B-Lymphozyten. Ebenso ist die Anzahl an Mastzellen (gesamt) wesentlich höher als die Summe Tryptase- und Chymase-positiver Mastzellen. Darüber hinaus wurden erstmals Chymase-positive Mastzellen im bovinen Endometrium immunhistologisch dargestellt, die in signifikant gerin-gerer Anzahl als Tryptase-positive Mastzellen im Endometrium vorkommen. Histologie und Zytologie wiesen vor allem bei den klinisch gesunden Rindern (Gruppe 2) und den sub-/infertilen Rindern (Gruppe 3) deutliche Diskrepanzen auf. Vorkommen, Grad und Charakter einer Endometritis konnten durch die Uteruszytologie nicht eindeutig bestimmt werden. Zum einen wurde bei histologisch unauffälligen Endometrien mittels Zytologie die Diagnose ‚subklinische’ Endometritis gestellt. Zum anderen blieben histologisch festgestellte, insbesondere chronische nicht-eitrige Endometritiden, in der Zytologie unerkannt. Somit soll-te die Beurteilung der Uteruszytologie nur unter Berücksichtigung klinischer Ergebnisse so-wie unter Einbeziehung des Zyklusstandes erfolgen. Zudem sollten anhand der eigenen Er-gebnisse die Vorteile und die Aussagekraft dieses Verfahrens überdacht werden. Im Gegensatz zu den Ergebnissen der zytologischen Untersuchung konnten mittels Endo-metriumbiopsie auch chronische nicht-eitrige Endometritiden sowie degenerative endo-metriale Veränderungen (Endometrose, Angiosklerose) erkannt werden. Somit stellt die En-dometriumbiopsie, besonders bei „repeat breedern“ und bei Kühen mit unklarer klinischer Symptomatik, eine sinnvolle Ergänzung der klinischen Diagnostik dar, um klinisch nicht er-fassbare endometriale Erkrankungen festzustellen. Die Entnahme einer Endometriumbiopsie hat keine negativen Auswirkungen auf die prospektive Reproduktionsleistung bei Milchkühen. Eine hochfrequente Entnahme von Endometriumbioptaten über einen Zyklus hinweg ist dagegen nicht empfehlenswert. Beim Rind stellen geringgradige endometriale Veränderungen mit einer erneuten Gravidität vereinbare Befunde dar, während mittel- und hochgradige Veränderungen die Prognose für eine erneute Trächtigkeit stark herabsetzen.

Master of Science / Department of Animal Sciences and Industry / Timothy G. Rozell / Several alternatively-spliced mRNA transcripts of the follicle stimulating hormone receptor (FSHR) have been identified in sheep, including FSHR-1 (G protein-coupled form), FSHR-2 (dominant negative form), and FSHR-3 (growth factor type-1 form). Coupling of the FSHR to signaling pathways which activate different downstream effectors leads to speculation that specific splice variants may be transcribed under differing physiological conditions. This is the first study to correlate expression patterns of FSHR-1, FSHR-2, and FSHR-3 and development of follicles in the mature sheep ovary. In Experiment 1, 8 Suffolk-cross ewes were allowed to come into estrus naturally and were euthanized 24 (n=3), 36 (n=3), and 48 (n=2) hours after the onset of estrus. In Experiment 2, 7 Suffolk-cross ewes received CIDRs for 14 days. At CIDR removal, PMSG (500IU) was administered to treatment ewes (n=3), while controls (n=4) received no PMSG. Ewes were euthanized 24 (n=4; 2 CIDR only, 2 PMSG) or 36 (n=3; 2 CIDR only, 1 PMSG) hours later. All visible follicles were aspirated and pooled according to follicular diameter: small (≤ 2.0 mm), medium (2.1-4.0 mm), large (4.1-6.0 mm), and preovulatory (≥ 6.1 mm). Granulosa cells were separated from follicular fluid by centrifugation. Total RNA was extracted from granulosa cells (GC) and reversed transcribed. The resulting cDNA was subjected to qPCR, using primer sets designed to amplify each variant specifically. For Experiment 1, regardless of time after onset of estrus, relative expression of FSHR-3 exceeded that of both FSHR-1 and FSHR-2 in medium follicles (p < 0.01), and tended to be higher in small follicles (p=0.09). For Experiment 2, treatment with PMSG did not significantly alter expression patterns of FSHR variants (p=0.18). The FSHR-3 was expressed higher than FSHR-2 in all follicle sizes (p < 0.01) and was numerically more highly expressed than FSHR-1, although this difference was not significant (p > 0.11). These experiments show that in addition to the well characterized G protein-coupled form of the FSHR, alternatively spliced variants of the FSHR may participate in follicular dynamics during the first follicular wave of the sheep estrous cycle. Furthermore, these results would indicate that an “alternatively” spliced form of the FSHR (FSHR-3) is the predominant form of the FSHR in the sheep.