Human immunodeficiency virus (HIV) infects 30 million people worldwide. In sub-Saharan Africa, the region most affected by HIV, women comprise 60% of the infected population. Heterosexual transmission is a major mode of viral acquisition, mandating further research of the process and prevention of HIV acquisition via the female reproductive tract (FRT). The FRT is a dynamic environment, protected by host immune mechanisms and commensal microbes. The disruption of either of these elements can increase susceptibility to HIV. Accordingly, one common risk factor for HIV acquisition is the microbial shift condition known as bacterial vaginosis (BV), which is characterized by the displacement of healthy lactobacilli by an overgrowth of pathogenic bacteria. As the bacteria responsible for BV pathogenicity and their interactions with host immunity are not understood, we sought to evaluate the effects of BV-associated bacteria on reproductive epithelia. Here we have characterized the interaction between BV-associated bacteria and the female reproductive tract by measuring cytokine and defensin induction in FRT epithelial cells following bacterial inoculation. Four BV-associated bacteria were evaluated alongside six lactobacilli for a comparative assessment. Our model showed good agreement with clinical BV trends; we observed a distinct cytokine and human β- defensin-2 response to BV-associated bacteria, especially Atopobium vaginae, compared to most lactobacilli. One lactobacillus species, Lactobacillus vaginalis, induced an immune response similar to that elicited by BV-associated bacteria. These iii data provide an important prioritization of BV-associated bacteria and support further characterization of reproductive bacteria and their interactions with host epithelia. We next evaluated the effect of this interaction on HIV infection by investigating the soluble effectors secreted when FRT epithelial cells were cocultured with A. vaginae. We observed increased proviral activity mediated by secreted low molecular weight effectors, and determined that this activity was not likely mediated by cytokine responses. Instead, we identified a complex mixture containing several upregulated host proteins. Selected individual proteins from the mixture exhibited HIV-enhancing activity only when applied with the complex mixture of proviral factors, suggesting that HIV enhancement might be mediated by synergistic effects. In addition to characterizing the immune interactions that mediate the enhanced HIV acquisition associated with BV, we also evaluated the safety and efficacy of RC- 101, a candidate vaginal microbicide being developed for the prevention of HIV transmission. RC-101 has been effective and well tolerated in preliminary cell culture and macaque models. However, the effect of RC-101 on primary vaginal tissues and resident vaginal microflora requires further evaluation. Here, we treated primary vaginal tissues and vaginal bacteria, both pathogenic and commensal, with RC-101 to investigate compatibility of this microbicide with FRT tissue and microflora. RC-101 was well tolerated by host tissues and commensal vaginal bacteria, while BV-associated bacteria were inhibited by RC-101. By establishing vaginal microflora, the specific antibacterial activity of RC-101 may provide a dual mechanism of HIV protection.

The F-strain of Mycoplasma gallisepticum (FMG) is commonly used in vaccination programs to displace infections by more virulent natural or wild type Mycoplasma gallisepticum strains. However, a better understanding of the mechanisms responsible for altered egg production (EP) and egg quality in commercial layers infected with FMG is important, as these alterations can cause economic loss to the United States layer industry. This study was designed to examine potential mechanism(s) responsible for alterations in EP and egg quality by FMG-inoculation. The effects of FMG on production parameters and physiological characteristics of commercial laying hens were evaluated. In isolation units, 12 wk FMG inoculation delayed onset of lay approximately one wk, decreased overall EP, and decreased EP 34 wk post-inoculation. A 12 wk FMG inoculation also resulted in a higher incidence of fatty liver hemorrhagic syndrome, ovarian follicular regression, and decreased isthmal and vaginal proportions of the reproductive tract. Ovarian regression may be related to retarded production (liver), transport (blood), and/or uptake (ovary) of yolk particles. Changes in blood characteristics (i.e. lipid composition) with FMG colonization of the liver may become manifest through changes in egg constituents. As evidenced through changes in the relative weights of various reproductive organs, colonization of these organs by FMG, in addition to the liver, may also be a cause of the effects observed on EP. Increases in hematocrit, serum triglycerides, and plasma protein between 8 and 10 wk post FMG-inoculation, suggest that the initial weeks of EP are stressful. Post-peak decreases in these same variables suggest a more chronic inhibition on lipid and protein synthesis in the liver. Decreased blood lipid concentration may be directly responsible for the reductions in yolk lipid, cholesterol, and fatty acid deposition in 12 wk FMG-inoculated hens. Dual adverse effects in the caged layer facility on feed conversion and egg mass were realized in 22 wk FMG-inoculated birds. In contrast, a 12 wk FMG inoculation delayed onset of lay without a loss in total EP or egg mass. Therefore, inoculation with FMG at 12 wk is more practical and cost effective. Higher degrees of physiological stress experienced by hens in a caged layer facility may exacerbate the effects of FMG inoculation seen in the isolation units. These data demonstrate that alterations in performance and egg characteristics of commercial layers inoculated with FMG at either 12 or 22 wk of age and housed in either isolation units or caged layer facilities are related to mutual functional disturbances in the blood, liver, ovary, and oviduct without concomitant intestinal changes.

Antimicrobial peptides (AMPs) are small proteins produced by epithelial surfaces, which have broad-spectrum antimicrobial and immunomodulatory activities. In the lung, skin and alimentary tract AMPs are known to be important in infectious and inflammatory conditions. Far less is known regarding the role of AMPs within the female reproductive tract, but as infection and inflammation are causes of preterm labour, AMPs may have a key function in maintain and protecting pregnancy. The major groups of human AMPs include the human beta defensins (HBDs), two antileukoproteinases (secretory leukocyte protease inhibitor (SLPI) and Trappin-2/Elafin), and the human cathelicidin hCAP18/LL-37, with several studies identifying their presence at sites throughout the reproductive tract. The cervix in pregnancy is positioned between the upper genital tract containing the developing fetus and the lower tract where infections usually arise. I hypothesise that AMPs are fundamental to mucosal immune defence of the cervix in pregnancy, preventing ascending infection and excessive inflammation that can cause preterm labour. This thesis focused on the human cathelicidin hCAP18/LL-37 and its role within the cervix and vagina. The aims of this thesis were to; investigate the inflammatory effects of LL-37 from cervical and vaginal derived epithelial cells and determine the pathways and receptors in which LL-37 may elicit its effects and how production may be regulated; investigate the role of CRAMP in a mouse model of preterm birth; and determine the production of AMPs by the pregnant cervix whilst investigating the relationship between AMP concentrations in cervicovaginal secretions and preterm labour. The inflammatory effect of LL-37 was investigated using cell lines derived from endocervical, ectocervical and vaginal epithelium. The study of these cell lines suggests divergent responses of cervical and vaginal epithelial cells. LL-37 mediated induction of IL-8 and IL-6 production from endocervical epithelial cells was observed in a dose-dependent and time-dependent manner, whilst ectocervical and vaginal cells also respond to treatment with LL-37 through IL-8 and IL-6 production. To determine a possible mechanism of action of LL-37 on IL-8 and IL-6 in the three cell lines, inhibitors against MAPK cascades, ERK, p38 MAPK and JNK, and known LL-37 receptors were investigated. In endocervical cells LL-37 mediated IL-8 occurs via activation of unidentified GPCRs, whilst in ectocervical cells this effect on IL‐8 and IL-6 is via the activation of ERK and p38 MAPK cascades. The mechanism by which LL-37 induces IL-8 secretion in vaginal epithelial cells remains unknown. Expression of LL-37 was shown to be mediated by vitamin D3 in vitro in cervical and vaginal epithelial cells. However when this relationship was investigated in vivo, using matched serum and cervicovaginal secretions from woman at early pregnancy, no correlation was observed between circulating vitamin D and cervicovaginal or circulating hCAP18/LL-37. However, the majority of women in this study reported with insufficient levels of vitamin D, which may effect the relationship observed with hCAP18/LL-37. Using a mouse model of LPS-induced preterm labour, to mimic the presence of intrauterine infection bacterial infection, I aimed to characterise the role of CRAMP, the mouse orthologue of hCAP18/LL-37, in the lower inflammatory and immune response that results in preterm labour. Wild type C57Bl/6J mice receiving an intrauterine injection of LPS deliver prematurely, within 24 hours of injection. However mice deficient in CRAMP (Camp -/-) receiving an intrauterine injection of LPS deliver significantly later and have a non-significant increase in pup survival compared to wild type C57Bl/6J mice. Cervical tissue collected post partum showed no difference in inflammatory markers between wild type C57Bl/6J and Camp -/- mice, however there was increased expression of the neutrophil chemoattractant marker, Cxcl5, and the neutrophil marker, Ngp in Camp -/- mice. In the lower genital tract, levels of antimicrobial peptides were determined in samples of cervicovaginal secretions collected from pregnant women. AMPs, hCAP18/LL-37, HBD-2 and SLPI were found in cervicovaginal secretions, and levels of hCAP18/LL-37 were increased in women with the common vaginal infection bacterial vaginosis. However no relationship was identified between the concentration of AMPs and preterm birth in this study. This work has shown that the lower genital tract, where infections that are associated with preterm labour originate, expresses the human cathelicidin hCAP18/LL-37. It may play an important role in modulating the immune response to invading infection associated with preterm labour. Further investigation of these responses may increase understanding of the physiology and pathophysiology of labour, and lead to strategies for the prevention of premature delivery.

<p>The vervet monkey, Chlorocebus aethiops, an Old World monkey, has been often used in biomedical research programs (toxicological studies and fertility) because of the inaccessibility of relevant human tissues. Data from nonhuman primates have been a vital component of advances in areas such as infertility, contraception, and other reproductive processes because of the phylogenetic closeness of the primates to humans. The aims and objectives of the study were 1) to describe the gross morphology, histology and ultrastructure of the male reproductive system, 2) to describe and compare the processes of spermatogenesis and spermiogenesis of the vervet monkey to humans and other nonhuman primates, and 3) to evaluate the vervet monkey as a possible experimental model for future human reproductive studies.</p> <p>Twenty-nine adult male vervet monkeys, aged between 5 and 11 years, were used. Gross morphological features of different organs of the reproductive tract were recorded. Light and electron microscopic techniques, and methacrylate sections were used on selected tissues of the reproductive tract. The results showed that the vervet monkey has a male&nbsp / reproductive system similar to many non-human primates studied and man. The epididymis was distinctively subdivided into the caput, corpus, and the caudal regions. No significant differences were observed on the epithelial height of these three regions. Four cell types, apical, principal, and basal cells, and the intraepithelial lymphocytes were observed. The basal cell distribution showed significant differences among three regions of the epididymis (P &le / 0.01). There were numerous phagocytic vesicles found in three regions of the epididymis. The Sertoli cells showed perforated sleeve-like processes which encased elongated and mature spermatids ready for spermiation. The nuclei of the Sertoli cells were found to be multilobed (4 to 5) compared to the less lobular nuclei of the human Sertoli cells (2 to 3). The Leydig cells showed typical features of steroidogenic cells with abundant smooth endoplasmic reticulum, numerous large mitochondria, and few rough endoplasmic reticulum.</p> <p>It was concluded that the gross morphology and structure of the reproductive tract of the vervet monkey has many similarities to humans and other mammals. Secondly, the organization of spermatogenesis is similar to that found in humans, and is commonly known as a helical arrangement. The results further suggest that the vervet monkey could be regarded as suitable model for human male reproductive studies.</p> <p>&nbsp / </p>

La aplicación de procesos biotecnológicos como la separación de espermatozoides X e Y mediante citometría de flujo, pueden generar la eliminación de determinados componentes del plasma seminal necesarios para conservar su funcionalidad. En este sentido, se ha observado que la adición de plasma seminal al medio espermático, protege a los espermatozoides cuando estos son sometidos a separación espermática mediante citometría de flujo. Sin embargo, diversos estudios demuestran que rel efecto beneficioso que ejerce el plasma seminal sobre los espermatozoides reside en determinadas proteínas. En el caso de la especie porcina, hay estudios previos que determinan que el efecto protector que ejerce el plasma seminal sobre los espermatozoides, se debe a una proteína denominada heterodímero PSP-I/PSP-II. El efecto beneficioso de dicha proteína sobre la funcionalidad espermática en espermatozoides de verraco altamente diluidos parece estar conservada, en su subunidad PSP-II y, concretamente, en la fracción peptídica de ésta. Estudiar, además, su localización a lo largo del tracto genital del verraco así como en las distintas fracciones del eyaculado, puede ser importante para llegar a conocer si la presencia de este heterodímero en el medio de recogida espermático de espermatozoides X e Y, es beneficioso tanto en la funcionalidad como en la capacidad fecundante de estos espermatozoides. / Biotechnological procedures of semen, such as sexing using flow cytometry/cell sorting procedures, causes high dilutions during sperm manipulation, linked with the wash away or high dilution of seminal plasma components. Thus, to develop strategies to extend the viability of treated spermatozoa are necessary. It is well known that add seminal plasma (SP) to the sperm media contributes to preserving the integrity and the fertilizing potential of sperm. Nevertheless, the beneficial effects of seminal plasma seem to be restricted to specific proteins of the SP. In porcine, previous studies have related this protective effect of the seminal plasma on the sperm cells to a protein called PSP-I/PSP-II heterodimer. The beneficial effect of this protein on the functionality of highly diluted boar spermatozoa is largely preserved in its PSP-II subunit and does not appear to require its glycan moiety. Moreover, study its localization along male reproductive tract and in different portions of the ejaculate could be important to know if the presence of PSP-I/PSP-II heterodimer in the collection medium for sex sorted spermatozoa is beneficial on the in vitro function and in vivo fertilizing ability.

<p>The vervet monkey, Chlorocebus aethiops, an Old World monkey, has been often used in biomedical research programs (toxicological studies and fertility) because of the inaccessibility of relevant human tissues. Data from nonhuman primates have been a vital component of advances in areas such as infertility, contraception, and other reproductive processes because of the phylogenetic closeness of the primates to humans. The aims and objectives of the study were 1) to describe the gross morphology, histology and ultrastructure of the male reproductive system, 2) to describe and compare the processes of spermatogenesis and spermiogenesis of the vervet monkey to humans and other nonhuman primates, and 3) to evaluate the vervet monkey as a possible experimental model for future human reproductive studies.</p> <p>Twenty-nine adult male vervet monkeys, aged between 5 and 11 years, were used. Gross morphological features of different organs of the reproductive tract were recorded. Light and electron microscopic techniques, and methacrylate sections were used on selected tissues of the reproductive tract. The results showed that the vervet monkey has a male&nbsp / reproductive system similar to many non-human primates studied and man. The epididymis was distinctively subdivided into the caput, corpus, and the caudal regions. No significant differences were observed on the epithelial height of these three regions. Four cell types, apical, principal, and basal cells, and the intraepithelial lymphocytes were observed. The basal cell distribution showed significant differences among three regions of the epididymis (P &le / 0.01). There were numerous phagocytic vesicles found in three regions of the epididymis. The Sertoli cells showed perforated sleeve-like processes which encased elongated and mature spermatids ready for spermiation. The nuclei of the Sertoli cells were found to be multilobed (4 to 5) compared to the less lobular nuclei of the human Sertoli cells (2 to 3). The Leydig cells showed typical features of steroidogenic cells with abundant smooth endoplasmic reticulum, numerous large mitochondria, and few rough endoplasmic reticulum.</p> <p>It was concluded that the gross morphology and structure of the reproductive tract of the vervet monkey has many similarities to humans and other mammals. Secondly, the organization of spermatogenesis is similar to that found in humans, and is commonly known as a helical arrangement. The results further suggest that the vervet monkey could be regarded as suitable model for human male reproductive studies.</p> <p>&nbsp / </p>

MARTINS, Jorge André Matias. Avaliação da biometria testicular, epididimal e de glândulas sexuais acessórias e correlações entre características biométricas e histológicas em carneiros deslanados sprd. 2006. 56 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Zootecnia, Fortaleza-CE, 2006. / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-08-01T12:33:32Z No. of bitstreams: 1 2006_dis_jammartins.pdf: 532644 bytes, checksum: 997a7780ea614be6ad84b2772b07b79a (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-08-01T12:33:59Z (GMT) No. of bitstreams: 1 2006_dis_jammartins.pdf: 532644 bytes, checksum: 997a7780ea614be6ad84b2772b07b79a (MD5) / Made available in DSpace on 2016-08-01T12:33:59Z (GMT). No. of bitstreams: 1 2006_dis_jammartins.pdf: 532644 bytes, checksum: 997a7780ea614be6ad84b2772b07b79a (MD5) Previous issue date: 2006 / The present study was conducted to evaluate aspects of the reproductive tract of crossbred rams raised in the semi-arid region of the Brazilian Northeast. Correlations between measurements of the testes, epididymis and accessory sex glands and variables related to the spermatogenic process were also determined. Data were obtained from 19 animals slaughtered at the approximate age of 60 weeks. Seminiferous tubules were studied with regard to volume, length and height and the population of Sertoli and germ cells. Associations between variables were estimated by Pearson’s method (p < 0.05) and linear regression analysis using StatView 5.0 software (SAS, 1998). Significant correlations were found among all testis measurements (weight, length, width and volume) and other aspects of the reproductive trait, such as length of the cauda epididymis (CCauEp; r = 0.66 to 0.68) and weight of the accessory sex glands (AG; r = 0.68 to 0.72). Weight of AG was also associated with epididymal weight and CCauEp (r = 0.56 and 0.70, respectively), suggesting the existence of a synchronized development of the male reproductive trait. Testis width (TW) represented the variable with the most significant correlations with testis weight, length and volume (r = 0.88; 0.87 and 0.94). Moreover, regression model allowed the empirical estimate of testis volume (TV) based on testis width: TV = - 260.17 + 68.479 x TW (R² = 0.95). This characteristic (TW) was highly correlated with important aspects of gonad histology, such as volume of the seminiferous tubules (r = 0.96), number of round spermatids per testis (r = 0.89) and daily production of round spermatids (r = 0.89). Linear regression models with high coefficients of determination (R²) allowed the estimation of histological parameters based on testis width. In conclusion, the present study shows, for the first time, information about the weight of accessory sex glands in crossbred rams. It was also observed that animals with larger testes had larger epididymis and accessory sex glands and greater quantitative estimates of the spermatogenesis. Testis width should be useful for selection of sires because it is of easy measurement. / O objetivo deste estudo foi avaliar a biometria de partes do trato reprodutor de ovinos deslanados sem padrão racial definido (SPRD) criados no semi-árido nordestino, e correlacionar os dados biométricos testiculares, epididimais e de glândulas sexuais acessórias com variáveis associadas ao processo espermatogênico. Os dados biométricos foram obtidos a partir dos testículos, epidídimos e das glândulas sexuais acessórias de 19 animais abatidos em torno de 60 semanas de idade. O parênquima testicular foi avaliado quanto ao diâmetro, volume e comprimento dos túbulos seminíferos, altura do epitélio germinativo e população de células e Sertoli e germinativas. Foram realizadas análises de correlação de Pearson (p < 0.05) entre as características estudadas e montados modelos de regressão linear simples para estas mesmas variáveis através do programa estatístico StatView, 5.0 (SAS, 1998). Correlações significativas foram verificadas entre todas as medidas testiculares (peso, comprimento, largura e volume) e as outras medidas do trato reprodutor, como comprimento da cauda do epidídimo (CCauEp; r=0,66 a 0,68) e as peso das glândulas sexuais acessórias (r=0,68 a 0,72). Estas também tiveram correlações significativas com o peso do epidídimo e com o CCauEp (r=0,56 e 0,70, respectivamente), o que sugere a existência de uma sinergia no desenvolvimento do trato genital masculino. A largura testicular (LT) foi o parâmetro que apresentou correlações mais significativas com peso, comprimento e volume testiculares (r=0,88; 0,87 e 0,94) e os modelos de regressão que permitem estimar qualquer uma dessas características, a exemplo do volume testicular: VT = -260,17 + 68,479 x LT; R²= 0,95. Esta característica também esteve alta e significativamente correlacionada com mportantes parâmetros histológicos, como volume do túbulo seminífero (r=0,96), número de espermátides arredondadas por testículo (r=0,89) e produção diária de espermátides arredondadas (r=0,89). Os modelos de regressão linear simples com coeficientes de determinação (R²) elevados permitem estimar as características histológicas a partir da largura testicular. No presente estudo, foram obtidos, pela primeira vez, os pesos das glândulas vesiculares e bulbo-uretrais em ovinos SPRD e pôde-se observar que animais com maior medidas testiculares também apresentaram elevados parâmetros quantitativos da espermatogênese e maiores epidídimos e glândulas vesiculares. A largura testicular pode ser útil para seleção de reprodutores por ser uma característica de fácil mensuração.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The experiment was conducted to evaluate the growth and reproductive development of beef heifers from 10 to 14 months of age in three food systems. Ryegrass pasture exclusively (Lolium multiflorum Lam.) or on pasture and receiving rice bran (0.8% of body weight) with or without ionophore. The experimental design was a completely randomized design with repeated measures, three food systems and nine replications, each animal considered an experimental unit. Body weight gain, body condition score and weight:height ratio (kg:cm) of heifers that received more ionophore rice bran were higher than those observed for heifers exclusively on ryegrass or receiving rice bran without ionophore. Regardless of the food system had hip height and pelvic area similar. Heifers that received rice bran with or without ionophore had higher reproductive tract score (RTS) that heifers exclusively on ryegrass. Heifers that received rice bran as a supplement with or without ionophore, the initial body weight of heifers was that best explained the variation in RTS, while the average daily gain of heifers on ryegrass exclusively explained the variation in RTS. / O experimento foi conduzido com o objetivo de avaliar o crescimento e o desenvolvimento reprodutivo de bezerras de corte dos 10 aos 14 meses em três sistemas alimentares, exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou em pastagem e recebendo farelo de arroz integral (FAI) com ou sem adição de ionóforo. A quantidade diária de FAI correspondeu a 0,8% do peso corporal. O delineamento experimental foi o inteiramente casualizado, com medidas repetidas no tempo, três sistemas alimentares e nove repetições, sendo cada animal considerado uma unidade experimental. O peso corporal, ganho de escore de condição corporal e de relação peso:altura (kg:cm) de bezerras que receberam FAI mais ionóforo foram superiores aos observados por bezerras exclusivamente em azevém ou recebendo FAI sem adição de ionóforo. Independente do sistema alimentar as bezerras apresentaram altura de garupa e área pélvica similar. Bezerras que receberam FAI com ou sem ionóforo apresentaram maior escore de trato reprodutivo (ETR) que bezerras exclusivamente em azevém. Para bezerras que receberam FAI como suplemento com ou sem adição de ionóforo, o peso corporal inicial foi que melhor explicou a variação em ETR, enquanto o ganho médio diário das bezerras exclusivamente em azevém explicou a variação em ETR.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O objetivo deste estudo foi avaliar a biometria de partes do trato reprodutor de ovinos deslanados sem padrÃo racial definido (SPRD) criados no semi-Ãrido nordestino, e correlacionar os dados biomÃtricos testiculares, epididimais e de glÃndulas sexuais acessÃrias com variÃveis associadas ao processo espermatogÃnico. Os dados biomÃtricos foram obtidos a partir dos testÃculos, epidÃdimos e das glÃndulas sexuais acessÃrias de 19 animais abatidos em torno de 60 semanas de idade. O parÃnquima testicular foi avaliado quanto ao diÃmetro, volume e comprimento dos tÃbulos seminÃferos, altura do epitÃlio germinativo e populaÃÃo de cÃlulas e Sertoli e germinativas. Foram realizadas anÃlises de correlaÃÃo de Pearson (p < 0.05) entre as caracterÃsticas estudadas e montados modelos de regressÃo linear simples para estas mesmas variÃveis atravÃs do programa estatÃstico StatView, 5.0 (SAS, 1998). CorrelaÃÃes significativas foram verificadas entre todas as medidas testiculares (peso, comprimento, largura e volume) e as outras medidas do trato reprodutor, como comprimento da cauda do epidÃdimo (CCauEp; r=0,66 a 0,68) e as peso das glÃndulas sexuais acessÃrias (r=0,68 a 0,72). Estas tambÃm tiveram correlaÃÃes significativas com o peso do epidÃdimo e com o CCauEp (r=0,56 e 0,70, respectivamente), o que sugere a existÃncia de uma sinergia no desenvolvimento do trato genital masculino. A largura testicular (LT) foi o parÃmetro que apresentou correlaÃÃes mais significativas com peso, comprimento e volume testiculares (r=0,88; 0,87 e 0,94) e os modelos de regressÃo que permitem estimar qualquer uma dessas caracterÃsticas, a exemplo do volume testicular: VT = -260,17 + 68,479 x LT; RÂ= 0,95. Esta caracterÃstica tambÃm esteve alta e significativamente correlacionada com mportantes parÃmetros histolÃgicos, como volume do tÃbulo seminÃfero (r=0,96), nÃmero de espermÃtides arredondadas por testÃculo (r=0,89) e produÃÃo diÃria de espermÃtides arredondadas (r=0,89). Os modelos de regressÃo linear simples com coeficientes de determinaÃÃo (RÂ) elevados permitem estimar as caracterÃsticas histolÃgicas a partir da largura testicular. No presente estudo, foram obtidos, pela primeira vez, os pesos das glÃndulas vesiculares e bulbo-uretrais em ovinos SPRD e pÃde-se observar que animais com maior medidas testiculares tambÃm apresentaram elevados parÃmetros quantitativos da espermatogÃnese e maiores epidÃdimos e glÃndulas vesiculares. A largura testicular pode ser Ãtil para seleÃÃo de reprodutores por ser uma caracterÃstica de fÃcil mensuraÃÃo. / The present study was conducted to evaluate aspects of the reproductive tract of crossbred rams raised in the semi-arid region of the Brazilian Northeast. Correlations between measurements of the testes, epididymis and accessory sex glands and variables related to the spermatogenic process were also determined. Data were obtained from 19 animals slaughtered at the approximate age of 60 weeks. Seminiferous tubules were studied with regard to volume, length and height and the population of Sertoli and germ cells. Associations between variables were estimated by Pearsonâs method (p < 0.05) and linear regression analysis using StatView 5.0 software (SAS, 1998). Significant correlations were found among all testis measurements (weight, length, width and volume) and other aspects of the reproductive trait, such as length of the cauda epididymis (CCauEp; r = 0.66 to 0.68) and weight of the accessory sex glands (AG; r = 0.68 to 0.72). Weight of AG was also associated with epididymal weight and CCauEp (r = 0.56 and 0.70, respectively), suggesting the existence of a synchronized development of the male reproductive trait. Testis width (TW) represented the variable with the most significant correlations with testis weight, length and volume (r = 0.88; 0.87 and 0.94). Moreover, regression model allowed the empirical estimate of testis volume (TV) based on testis width: TV = - 260.17 + 68.479 x TW (RÂ = 0.95). This characteristic (TW) was highly correlated with important aspects of gonad histology, such as volume of the seminiferous tubules (r = 0.96), number of round spermatids per testis (r = 0.89) and daily production of round spermatids (r = 0.89). Linear regression models with high coefficients of determination (RÂ) allowed the estimation of histological parameters based on testis width. In conclusion, the present study shows, for the first time, information about the weight of accessory sex glands in crossbred rams. It was also observed that animals with larger testes had larger epididymis and accessory sex glands and greater quantitative estimates of the spermatogenesis. Testis width should be useful for selection of sires because it is of easy measurement.

Orientadores: Luis Antonio Violin Dias Pereira, Lúcia Elvira Alvares / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-24T14:05:01Z (GMT). No. of bitstreams: 1 Barbieri_MainaraFerreira_M.pdf: 6077469 bytes, checksum: c80011c6f6fca680e92ac99691a9c348 (MD5) Previous issue date: 2014 / Resumo: A origem da albumina nos órgãos do trato reprodutor masculino tem sido questionada ao longo dos anos sendo sua presença tecidual constantemente atribuída a um mecanismo de extravasamento do plasma sanguíneo para o espaço extravascular intersticial, atribuindo-se ao fígado o papel de sítio de síntese desta proteína no período pós-natal. O padrão de distribuição da proteína albumina nos tecidos epiteliais de órgãos do trato reprodutor masculino reserva questionamentos quanto à veracidade de seu alcance luminal pelo rompimento de suas barreiras hematoteciduais. Carreador de esteroides; estimulador de células de Leydig; mediador do mecanismo parácrino de comunicação entre células de Sertoli e germinativas; promotor de proliferação celular e mediador de mudanças estruturais em membranas celulares, com suposta participação na reação acrossômica e capacitação do espermatozoide são possíveis papéis atribuídos à albumina no trato reprodutor masculino. A associação da proteína albumina a componentes glicídicos não é usual para mamíferos, porém há respaldo a essa hipótese através das PES (glicoproteínas pré-albumínicas epidídimo-específicas de ovinos) e dados de nosso grupo de pesquisa através do epítopo reconhecido pelo anticorpo monoclonal (Amc) TRA 54 (testicular germ cells immunized to rat). Essa dissertação de mestrado retoma a polêmica discussão centrada na presença, origem, localização e função da albumina no trato reprodutor masculino e se propôs, através de uma abordagem cronológica pós-natal (períodos neonatal, pré-púbere, púbere, adulto e idoso) a rediscutir os fundamentos teóricos associados à principal proteína sérica de mamíferos bem como o dúbio perfil de expressão dessa proteína em Ratos analbuminêmicos Nagase (NAR). Nesse estudo, a presença da proteína albumina foi evidenciada por immunoblotting em todos os grupos e períodos do desenvolvimento pós-natal, apresentando padrão distinto de bandeamento para testículos e epidídimos NAR. A localização celular da proteína albumina foi identificada em camundongos, por imunohistoquímica, principalmente no tecido intersticial e nas células da linhagem germinativa. Testículos de ratos Sprague Dawley adultos apresentaram marcação adicional para células de Sertoli. Testículos e epidídimos de NAR mantiveram reatividade intersticial, sugerindo, ainda que em níveis extremamente baixos, a expressão de albumina. Para epidídimos, foi identificado amplo espectro de perfis de imunomarcação das células epiteliais e estereocílios. Ductos deferentes apresentaram, na região proximal ao epidídimo, imunomarcação citoplasmática e, na região distal, marcação de estereocílios e espermatozoides luminais. Perfis de imunomarcação citoplasmática também foram observados em vesícula seminal, próstata e uretra peniana, principalmente, no período adulto. Por fim, a presença de transcritos de RNAm detectados por ensaios de RT PCR (reação em cadeia de polimerase via transcriptase reversa) para testículos e epidídimos foi confirmada desde o período neonatal até idoso, mostrando-se também presente para ducto deferente a partir da puberdade. A abordagem do desenvolvimento pós-natal revelou um pico de maior expressão da albumina no período adulto com início do declínio registrado no período idoso. Como ressalva final, a confirmação da presença de RNAm para órgãos do trato reprodutor masculino não invalida a hipótese da também presença de albumina circulante, sintetizada, em primeira instância, pelas células hepáticas / Abstract: The origin of the albumin in the male reproductive tract has been questioned over the years and their presence tissue constantly assigned to a mechanism for extravasation of plasma into the interstitial extravascular space, assigning the role of the liver as the site of synthesis of this protein in the postnatal period. The distribution pattern of the albumin protein in the epithelial tissues of organs of the male reproductive tract reserves inquiries as to the veracity of their luminal reach through the disruption of their tissue blood-barriers. Carrier steroids; stimulator of Leydig cells; mediator of paracrine mechanism of communication between Sertoli and germ cells; promoter of cell proliferation and mediator of structural changes in cell membranes, with alleged involvement in the acrosome reaction and capacitation of sperm are possible roles of albumin in the male reproductive tract. The association of the albumin protein to glycidyl components is unusual for mammals, but there is support to this hypothesis by the PES (prealbumins epididymis-specific glycoproteins of sheep) and data from our research group through the epitope recognized by the monoclonal antibody (mAb) TRA 54 (testicular germ cells immunized to rat). This dissertation takes up the controversial discussion centered on the presence, source, location and function of albumin in the male reproductive tract and proposed, through a chronological postnatal approach (neonatal, prepubertal, puberal, adult and elderly periods) to revisit the theoretical foundations associated with major serum protein of mammals as well as the dubious expression profile of this protein in Nagase analbuminemic rats (NAR). In this study, the presence of albumin protein was detected by immunoblotting in all groups and periods of postnatal development, with distinct banding pattern in testis and epididymis of NAR. The cellular localization of the albumin protein has been identified in mice by immunohistochemistry mainly in the interstitial tissue and in germline cells. Testes of Sprague Dawley rats showed additional labeling of Sertoli cells. Testes and epididymides of NAR maintained interstitial reactivity, suggesting the expression of albumin even at extremely low levels. To epididymis a broad spectrum of immunostaining of epithelial cells and stereocilia profiles were identified. Vas deferens showed cytoplasmic immunostaining in the region proximal to epididymis, and in the distal region, stereocilia and luminal spermatozoa labeling. Cytoplasmic immunostaining profiles were also observed in the seminal vesicle, prostate and penile urethra, especially in adult period. Finally, the presence of mRNA transcripts detected by RT PCR (reverse transcriptase polymerase chain reaction) assays to testis and epididymis was confirmed from neonates to the elderly, being also present in vas deferens from puberty. The approach of postnatal development revealed a peak with the highest expression of albumin in the adult period beginning to decline at elderly. As a final caveat, the presence of mRNA confirmed for organs of the male reproductive tract does not invalidate the hypothesis of the also presence of circulating albumin in reproductive tissue, which must be synthesized at first instance, by liver cells / Mestrado / Biologia Tecidual / Mestra em Biologia Celular e Estrutural