Orientador: Celso Antônio Rodrigues / Resumo: O comportamento das mulas, durante o estro, influencia negativamente sua função zootécnica. A ovariectomia tem se apresentado como forma de torná-las mais cooperativas. Procedimentos realizados por videocirurgia se apresentam com uma possibilidade para reduzir as complicações trans e pós-operatórias. Então, foram utilizadas 9 mulas, com idade de 7,9 ± 3,2 anos, pesando 352 ± 76 kg. Realizou-se estudo do comportamento reprodutivo dos animais durante a rufiação com garanhão equino, nas diferentes fases do ciclo estral. A duração do estro foi de 6,4 ± 1,4 dias. As ovulações ocorreram 5,3 ± 2,6 dias após o início dos sinais de estro. Os sinais que apresentaram maior correlação foram micção frequente (MF) e levantamento da cauda (LC) (0,79) e imobilidade (IM) e LC (0,76) enquanto que os que apresentaram menor correlação foram aproximação do macho (AM) e LC (0,26) e AM e MF (0,29). Posteriormente foi realizada a ovariectomia por videolaparoscopia e no pós-operatório foi realizado o acompanhamento clínico-laboratorial para caracterizar possíveis complicações da técnica. Para hemostasia dos vasos presentes no pedículo ovariano utilizou-se no grupo A um bisturi ultrassônico e no grupo B um aplicador de clips de laqueação de titânio. Observou-se que o grau de dificuldade da ovariectomia foi inversamente proporcional ao tamanho do ovário. O enfisema subcutâneo foi a complicação pós-operatória mais evidente, sendo observada em todo grupo (9/9), e o hemoperitôneo foi considerado a complic... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The mules’ behavior influences their zootecnical function negatively during the estrus. Ovariectomy is an interesting way to make these animals more cooperative. Videosurgeric procedures has become a possibility to reduce operative and postoperative complications. Nine mules underwent the procedure and were followed from January to May 2017. The average age of the animals was 7,9 ± 3,2 years old and the weight approximately 352 ± 76 kg. A prospective evaluation of reproductive behavior was set during the ruffling of the stallion equine, along different phases of the estrus. Estrus length was 6,4 ± 1,4 days. Ovulation occurred 5.3 ± 2.6 days after the onset of estrus signs. The signs that presented the highest correlation were frequent micturition (MF) with tail lifting (LC) (0,79) and immobility with just LC (0,76). The least correlations were male approximation (AM) with LC and AM with MF. Subsequently, video-laparoscopic ovariectomy was performed. In post-operative days, clinical follow up and laboratory assay were performed to detect possible complications. Two groups were randomly selected for ultrasonic scalpel (group A) and titanium clips (group B) aiming the hemostasis of the vascular pedicle on the ovaries. The difficulty of the procedure was greater as smaller the ovary was. Some complications were observed. Subcutaneous emphysema was found in all mules studied, possibly caused by CO2 blowing. On the other hand, hemo-peritoneum was uncommon (just one animal developed... (Complete abstract click electronic access below) / Doutor

Ziel der vorliegenden Arbeit war die ausführliche histomorphologische Charakterisierung der endometrialen Funktionsmorphologie sowie die Darstellung der Ki-67 Antigen, Östrogen- und Progesteronrezeptor-Expression am Endometrium zyklischer und azyklischer Maultierstuten unter Einbeziehung klinischer und endokrinologischer Daten. Die Ergebnisse sollten mit den zu diesem Thema bereits vorliegenden Erkenntnissen bei der Pferdestute verglichen werden. Zu diesem Zweck wurden 97 Endometriumbioptate von sechs klinisch-gynäkologisch gesunden Maultierstuten im Alter von 13 bis 23 Jahren (immun-)histologisch ausgewertet. Die Bioptate waren zuvor in Abständen von zwei bis zehn Tagen und über einen Zeitraum von 4 Monaten (April bis August 2004), im Fall einer Stute von sieben Monaten (April bis November 2004) entnommen worden. Für jedes der Tiere lagen detaillierte Informationen über die klinisch-gynäkologischen Befunde und die Serumhormonkonzentrationen zu den entsprechenden Entnahmezeitpunkten vor (BARTMANN et al. 2008). Vorberichtlich zeigen zwei der Tiere im Untersuchungszeitraum ein unregelmäßiges Zyklusgeschehen mit ovariellen Funktionskörpern sowie variablen Serumöstradiol- und –progesteronkonzentrationen, bei vier Maultieren wird klinisch eine Azyklie mit inaktiven Ovarien sowie niedrigen, wenig schwankenden Serumhormonwerten dokumentiert. Bei den Tieren der zyklischen Gruppe (n=2) variiert die endometriale Funktionsmorphologie sehr stark. In nahezu allen untersuchten Endometriumbioptaten findet sich eine ausgeprägte endometriale Fehldifferenzierung. Diese kann anhand ihres Erscheinungsbildes nach den bei der Pferdestute bekannten histomorphologischen Kriterien (HÄFNER 1999, SCHOON et al. 1999b) eingeteilt werden. Sie tritt bei den untersuchten Maultieren als irreguläre glanduläre oder aber als irreguläre und ungleichmäßige glanduläre Differenzierung in Erscheinung und wird mit einem inaktiven, sekretorischen oder proliferativen Grundcharakter vorgefunden. Maultierstute 1 zeigt im Untersuchungszeitraum eine variable endometriale Proliferationsaktivität (Marker Ki-67 Antigen), bei Maultierstute 2 ist sie konstant schwach ausgeprägt. Es lässt sich kein charakteristisches, zyklusphasenabhängiges, positiv an die Steroidhormonrezeptorexpression gekoppeltes Expressionsmuster, wie es bei der Pferdestute bekannt ist, nachweisen. Die ER- und PR-Expression zeigt bei den zyklischen Maultierstuten ein sehr variables Bild (insbesondere Maultierstute 1), welches dem bei zyklierenden Pferdestuten während der physiologischen Decksaison nicht entspricht. Aufgrund der Diskrepanz zwischen klinischen Daten, Serumhormonkonzentrationen, endometrialer Funktionsmorphologie und Hormonrezeptorstatus bei den zyklischen Maultierstuten kann festgestellt werden, dass es bei den untersuchten Tieren nicht möglich ist, anhand der morphologisch-funktionellen Gegebenheiten am Endometrium auf den klinischen Zyklusstand zu schließen und vice versa. Die erhobenen Befunde sind in ihrer Gesamtheit am ehesten vergleichbar mit den Verhältnissen bei Pferdestuten in den Übergangszyklen zwischen Winteranöstrus und physiologischer Decksaison. Bei der Gruppe der azyklischen Maultiere (n=4) findet sich in allen untersuchten Endometriumbioptaten eine inaktive endometriale Funktionsmorphologie. Diese passt grundsätzlich zu dem klinischen Vorbericht der Azyklie mit inaktiven Ovarien und niedrigen, wenig schwankenden Serumöstradiol- und -progesteronkonzentrationen. In der Mehrzahl der Bioptate liegt eine Irregularität der glandulären Epithelien vor und in einigen Fällen treten hyperplastische und hypersekretorische Drüsenareale auf. Diese Befunde können, anlehnend an die Erkenntnisse bei der Pferdestute, als Kennzeichen einer endometrialen Fehldifferenzierung interpretiert werden (HÄFNER 1999, SCHOON et al. 1999b). Im Zusammenhang mit der schwachen endometrialen Proliferationsaktivität und der variablen ER- und PR-Expression lässt sich eine deutliche Ähnlichkeit zu den Befunden bei Pferdestuten im Winteranöstrus bzw. im Anöstrus während der physiologischen Decksaison finden (AUPPERLE et al. 2003, GOCKELN et al. 2006). Die möglichen Ursachen der endometrialen Fehldifferenzierung bei den Maultierstuten können im Rahmen der durchgeführten Untersuchungen nicht abschließend geklärt werden. Es ist jedoch denkbar, dass, analog zur Pferdestute, eine ovariell oder zentral bedingte Dysregulation und/oder eine Störung auf der Effektor/Rezeptor-Ebene vorliegen können. Die Hauptursache für die Infertilität bei Maultierstuten besteht in den chromosomal bedingten Störungen im Ablauf der Meiose der Keimzellen (WODSEDALEK 1916). Aufgrund der eigenen Untersuchungsergebnisse kann jedoch angenommen werden, dass die diagnostizierte endometriale Fehldifferenzierung, die variable Steroidhormonrezeptor-Expression sowie das Vorliegen degenerativer endometrialer Alterationen, z.B. der Endometrose, in Anlehnung an die Erkenntnisse bei der Pferdestute, zusätzliche Faktoren darstellen könnten, die zur Unfruchtbarkeit der Maultiere beitragen. Die Ergebnisse der vorliegenden Arbeit beruhen jedoch lediglich auf der Untersuchung von sechs Maultierstuten und sollten aus diesem Grund nicht unkritisch auf die Gesamtpopulation der Maultiere übertragen werden. Weitere Verlaufsuntersuchungen an einem größeren Tiergut in verschiedenen Altersgruppen inkl. Betrachtung der Hypophysenhormone FSH und LH könnten zur Aufklärung der Ursache sowie der Bedeutung der endometrialen Fehldifferenzierung für die Infertilität beim Maultier beitragen. / The aim of the present study was a detailed histomorphological characterization of the functional morphology and the evaluation of Ki-67 antigen, estrogen and progesterone receptor expression patterns in the endometrium of cyclic and non-cyclic mule mares, with respect to clinical and endocrinological data. The results were to be compared with corresponding observations in the endometrium of horse mares. For this purpose 97 endometrial biopsies from six clinically and gynaecologically healthy mule mares, aged between 13 and 23 years, were evaluated histologically and immunohistochemically. The biopsies were taken in intervals from two up to ten days over a period of four months (April to August 2004), in one mare over seven months (April to November 2004). Detailed information was given about clinical and gynaecological findings. The serum hormone concentrations for each day of biopsy sampling were determined (BARTMANN et al. 2008). According to the clinical record two of the animals show an erratic estrous cycle associated with the occurrence of corpora lutea and follicles on the ovaries during the whole period of investigation. Additionally, variable plasma estrogen and progesterone values are found. Four mules reveal acyclia including inactive ovaries and constant low serum hormone levels. In the animals of the cyclic group (n=2) the endometrial functional morphology varies to a great extend. In almost every investigated endometrial biopsy a marked endometrial maldifferentiation can be detected. It is classified according to common histomorphological criteria used for the endometrium of horse mares (HÄFNER 1999, SCHOON et al. 1999b). Endometrial maldifferentiation of the mules investigated occurs as irregular glandular or irregular and unequal differentiation and appeares in combination with a proliferative, secretory or inactive functional morphology. Mule 1 shows variable and mule 2 constant low endometrial Ki-67 expression patterns during the period of investigation. In contrast to horses, no characteristic, stage of cycle- or steroid hormone receptor expression-dependent proliferation activity can be observed with the mule mares of the present study. The expression of estrogen and progesterone receptors is highly variable in the animals of the cyclic group (especially in mule 1). These findings do not correspond to the ones in horse mares during the physiological breeding season. Due to the discrepancy between clinical data, serum hormone values, endometrial functional morphology and hormone receptor expression in the investigated mules, a conclusion from the clinical stage of cycle to morpho-functional conditions in the endometrium and vice versa seems to be impossible. The results of the present study are most likely comparable with findings in horse mares during the transitional period between winter anestrus and the physiological breeding season. The mules of the acyclic group (n=4) reveal an inactive endometrial functional morphology in all investigated endometrial biopsies. This finding basically corresponds to the clinical record of acyclia associated with inactive ovaries and rather constant serum estrogen and progesterone concentrations. In the case of these animals a majority of the investigated endometria show irregular glandular epithelia. In some areas hyperplastic and hypersecretory glands can be seen. According to observations in horse mares these findings can be interpreted as signs of endometrial maldifferentiation (HÄFNER 1999, SCHOON et al. 1999b). In connection with the weak proliferation activity and the variable ER- and PR-expression patterns seen in the acyclic mules, these results resemble findings in horses during winter anestrus or anestrus during the breeding season (AUPPERLE et al. 2003, GOCKELN et al. 2006). Within the present study the possible reasons for endometrial maldifferentiation in mules could not be finally evaluated. But it seems possible that, like in horse mares, an ovarian or central dysregulation and/or dysfunction regarding the level of receptors and effectors could play a certain role. Infertility in she-mules is mainly due to the chromosomal abnormalities during meiosis of germ cells (WODSEDALEK 1916). According to the findings in horse mares, the results of this investigation indicate that the diagnosed endometrial maldifferentiation, the variable expression of steroid hormone receptors and the presence of endometrial degenerative lesions (e.g. endometrosis) could be additional factors that possibly lead to sterility in mule mares, too. Due to the small number of individuals examined in this study, the results should not uncritically be applied to the whole population of mules. Further clinical, histological and immunohistochemical investigations, including the pituitary hormones FSH and LH as well as the use of a larger number of animals at different stages of age, could lead to a better understanding of the causes of endometrial maldifferentiation and its relevance for the infertility in mules.

No description available.

A morfologia e a fisiologia ovariana nos equinos têm sido motivo de estudo devido a seu córtex e medula invertidos em comparação às das outras espécies mamíferas e seu ciclo reprodutivo fotoperíodo-dependente e à existência de uma fossa de ovulação. Estes aspectos levam ao profissional do campo da reprodução equina a se interessar em aprofundar cada vez mais no desenvolvimento de pesquisas que possam auxiliar oferecendo métodos e técnicas que ajudem no trabalho de campo. Uma das melhores técnicas utilizadas para o estudo da atividade ovariana têm sido as imagens ultrassonográficas nos seus diferentes modos de operação. Uma das estruturas ovarianas que se pode avaliar e acompanhar com o ultrassom é o Corpo Lúteo (CL), sendo ele vital pela produção de Progesterona (P4), hormônio encarregado da preparação do endométrio para oferecer um ambiente ao embrião. É relatada uma relação positiva entre quantidade de células lúteas e produção de P4. O objetivo deste estudo foi testar uma metodologia que permitisse avaliar a densidade celular por meio da histologia e determinar se existe relação com o numero de pixéis lúteos numa imagem de ultrassom e assim ter um dado confiável para tomar decisões. Foram coletados 29 ovários de éguas crioulas em diferentes fases do diestro, analisados todos dentro de um único grupo. As amostras foram passadas pelo ultrassom para obter a imagem ultrasonográfica salva diretamente do ultrassom. Posteriormente, os CLs foram dessecados para fazer laminas corada pela Hematoxilina Eosina. Os resultados mostraram que entre as variáveis existe correlação inversa de R=-0.61. O coeficiente de determinação achado foi de aproximadamente R2=40% com uma probabilidade P=0.04%. Os resultados indicam que, existem variações no numero de pixéis lúteos que podem explicar em grande percentagem os valores totais do numero de células presentes no tecido lúteo. / The ovarian morphology and physiology in horses has been the subject of study due to its reversed cortex and medulla in comparison to other mammalian species, the existance of an ovulation fossa and for its photoperiod-dependent reproductive cycle. These aspects lead to the field practitioner specialist on equine to develop research that may help at fieldwork. One of the best techniques for the study of ovarian activity has been the ultrassonographic images in its different modes of operation. The corpus luteum (CL) is one of the ovarian structures that can be assessed and followed by ultrasound, being vital for the production of progesterone (P4), hormone which prepares the endometrium to provide a good environment for embryo development. A positive relationship between amount of luteal cells and P4 production has been reported before. The aim of this study was to test a methodology that would assess cell density by histology and determine whether there is a correlation between the number of ultrasound image pixels and the luteal cells concentration and thus have a really trusted ultrasound data which might help veterinarians to make decisions. Twenty nine ovaries from criollo mares were collected at different stages of diestrus, and all analyzed within a unique group. Samples were assessed by ultrasound to obtain the ultrasonographic image. After that, the CLs were dried to make microscopy slides stained with hematoxylin-eosin. The results showed that an inverse correlation (r = -0.61) exists between the variables studied. The determination coefficient found was approximately R2 = 40% with a probability of P = 0.04%. The results indicate that there are variations in the number of pixels that can explain differences in the numbers of luteal cells in the corpus luteum tissue.

A morfologia e a fisiologia ovariana nos equinos têm sido motivo de estudo devido a seu córtex e medula invertidos em comparação às das outras espécies mamíferas e seu ciclo reprodutivo fotoperíodo-dependente e à existência de uma fossa de ovulação. Estes aspectos levam ao profissional do campo da reprodução equina a se interessar em aprofundar cada vez mais no desenvolvimento de pesquisas que possam auxiliar oferecendo métodos e técnicas que ajudem no trabalho de campo. Uma das melhores técnicas utilizadas para o estudo da atividade ovariana têm sido as imagens ultrassonográficas nos seus diferentes modos de operação. Uma das estruturas ovarianas que se pode avaliar e acompanhar com o ultrassom é o Corpo Lúteo (CL), sendo ele vital pela produção de Progesterona (P4), hormônio encarregado da preparação do endométrio para oferecer um ambiente ao embrião. É relatada uma relação positiva entre quantidade de células lúteas e produção de P4. O objetivo deste estudo foi testar uma metodologia que permitisse avaliar a densidade celular por meio da histologia e determinar se existe relação com o numero de pixéis lúteos numa imagem de ultrassom e assim ter um dado confiável para tomar decisões. Foram coletados 29 ovários de éguas crioulas em diferentes fases do diestro, analisados todos dentro de um único grupo. As amostras foram passadas pelo ultrassom para obter a imagem ultrasonográfica salva diretamente do ultrassom. Posteriormente, os CLs foram dessecados para fazer laminas corada pela Hematoxilina Eosina. Os resultados mostraram que entre as variáveis existe correlação inversa de R=-0.61. O coeficiente de determinação achado foi de aproximadamente R2=40% com uma probabilidade P=0.04%. Os resultados indicam que, existem variações no numero de pixéis lúteos que podem explicar em grande percentagem os valores totais do numero de células presentes no tecido lúteo. / The ovarian morphology and physiology in horses has been the subject of study due to its reversed cortex and medulla in comparison to other mammalian species, the existance of an ovulation fossa and for its photoperiod-dependent reproductive cycle. These aspects lead to the field practitioner specialist on equine to develop research that may help at fieldwork. One of the best techniques for the study of ovarian activity has been the ultrassonographic images in its different modes of operation. The corpus luteum (CL) is one of the ovarian structures that can be assessed and followed by ultrasound, being vital for the production of progesterone (P4), hormone which prepares the endometrium to provide a good environment for embryo development. A positive relationship between amount of luteal cells and P4 production has been reported before. The aim of this study was to test a methodology that would assess cell density by histology and determine whether there is a correlation between the number of ultrasound image pixels and the luteal cells concentration and thus have a really trusted ultrasound data which might help veterinarians to make decisions. Twenty nine ovaries from criollo mares were collected at different stages of diestrus, and all analyzed within a unique group. Samples were assessed by ultrasound to obtain the ultrasonographic image. After that, the CLs were dried to make microscopy slides stained with hematoxylin-eosin. The results showed that an inverse correlation (r = -0.61) exists between the variables studied. The determination coefficient found was approximately R2 = 40% with a probability of P = 0.04%. The results indicate that there are variations in the number of pixels that can explain differences in the numbers of luteal cells in the corpus luteum tissue.

A morfologia e a fisiologia ovariana nos equinos têm sido motivo de estudo devido a seu córtex e medula invertidos em comparação às das outras espécies mamíferas e seu ciclo reprodutivo fotoperíodo-dependente e à existência de uma fossa de ovulação. Estes aspectos levam ao profissional do campo da reprodução equina a se interessar em aprofundar cada vez mais no desenvolvimento de pesquisas que possam auxiliar oferecendo métodos e técnicas que ajudem no trabalho de campo. Uma das melhores técnicas utilizadas para o estudo da atividade ovariana têm sido as imagens ultrassonográficas nos seus diferentes modos de operação. Uma das estruturas ovarianas que se pode avaliar e acompanhar com o ultrassom é o Corpo Lúteo (CL), sendo ele vital pela produção de Progesterona (P4), hormônio encarregado da preparação do endométrio para oferecer um ambiente ao embrião. É relatada uma relação positiva entre quantidade de células lúteas e produção de P4. O objetivo deste estudo foi testar uma metodologia que permitisse avaliar a densidade celular por meio da histologia e determinar se existe relação com o numero de pixéis lúteos numa imagem de ultrassom e assim ter um dado confiável para tomar decisões. Foram coletados 29 ovários de éguas crioulas em diferentes fases do diestro, analisados todos dentro de um único grupo. As amostras foram passadas pelo ultrassom para obter a imagem ultrasonográfica salva diretamente do ultrassom. Posteriormente, os CLs foram dessecados para fazer laminas corada pela Hematoxilina Eosina. Os resultados mostraram que entre as variáveis existe correlação inversa de R=-0.61. O coeficiente de determinação achado foi de aproximadamente R2=40% com uma probabilidade P=0.04%. Os resultados indicam que, existem variações no numero de pixéis lúteos que podem explicar em grande percentagem os valores totais do numero de células presentes no tecido lúteo. / The ovarian morphology and physiology in horses has been the subject of study due to its reversed cortex and medulla in comparison to other mammalian species, the existance of an ovulation fossa and for its photoperiod-dependent reproductive cycle. These aspects lead to the field practitioner specialist on equine to develop research that may help at fieldwork. One of the best techniques for the study of ovarian activity has been the ultrassonographic images in its different modes of operation. The corpus luteum (CL) is one of the ovarian structures that can be assessed and followed by ultrasound, being vital for the production of progesterone (P4), hormone which prepares the endometrium to provide a good environment for embryo development. A positive relationship between amount of luteal cells and P4 production has been reported before. The aim of this study was to test a methodology that would assess cell density by histology and determine whether there is a correlation between the number of ultrasound image pixels and the luteal cells concentration and thus have a really trusted ultrasound data which might help veterinarians to make decisions. Twenty nine ovaries from criollo mares were collected at different stages of diestrus, and all analyzed within a unique group. Samples were assessed by ultrasound to obtain the ultrasonographic image. After that, the CLs were dried to make microscopy slides stained with hematoxylin-eosin. The results showed that an inverse correlation (r = -0.61) exists between the variables studied. The determination coefficient found was approximately R2 = 40% with a probability of P = 0.04%. The results indicate that there are variations in the number of pixels that can explain differences in the numbers of luteal cells in the corpus luteum tissue.

Ziel der vorliegenden Arbeit war die ausführliche histomorphologische Charakterisierung der endometrialen Funktionsmorphologie sowie die Darstellung der Ki-67 Antigen, Östrogen- und Progesteronrezeptor-Expression am Endometrium zyklischer und azyklischer Maultierstuten unter Einbeziehung klinischer und endokrinologischer Daten. Die Ergebnisse sollten mit den zu diesem Thema bereits vorliegenden Erkenntnissen bei der Pferdestute verglichen werden. Zu diesem Zweck wurden 97 Endometriumbioptate von sechs klinisch-gynäkologisch gesunden Maultierstuten im Alter von 13 bis 23 Jahren (immun-)histologisch ausgewertet. Die Bioptate waren zuvor in Abständen von zwei bis zehn Tagen und über einen Zeitraum von 4 Monaten (April bis August 2004), im Fall einer Stute von sieben Monaten (April bis November 2004) entnommen worden. Für jedes der Tiere lagen detaillierte Informationen über die klinisch-gynäkologischen Befunde und die Serumhormonkonzentrationen zu den entsprechenden Entnahmezeitpunkten vor (BARTMANN et al. 2008). Vorberichtlich zeigen zwei der Tiere im Untersuchungszeitraum ein unregelmäßiges Zyklusgeschehen mit ovariellen Funktionskörpern sowie variablen Serumöstradiol- und –progesteronkonzentrationen, bei vier Maultieren wird klinisch eine Azyklie mit inaktiven Ovarien sowie niedrigen, wenig schwankenden Serumhormonwerten dokumentiert. Bei den Tieren der zyklischen Gruppe (n=2) variiert die endometriale Funktionsmorphologie sehr stark. In nahezu allen untersuchten Endometriumbioptaten findet sich eine ausgeprägte endometriale Fehldifferenzierung. Diese kann anhand ihres Erscheinungsbildes nach den bei der Pferdestute bekannten histomorphologischen Kriterien (HÄFNER 1999, SCHOON et al. 1999b) eingeteilt werden. Sie tritt bei den untersuchten Maultieren als irreguläre glanduläre oder aber als irreguläre und ungleichmäßige glanduläre Differenzierung in Erscheinung und wird mit einem inaktiven, sekretorischen oder proliferativen Grundcharakter vorgefunden. Maultierstute 1 zeigt im Untersuchungszeitraum eine variable endometriale Proliferationsaktivität (Marker Ki-67 Antigen), bei Maultierstute 2 ist sie konstant schwach ausgeprägt. Es lässt sich kein charakteristisches, zyklusphasenabhängiges, positiv an die Steroidhormonrezeptorexpression gekoppeltes Expressionsmuster, wie es bei der Pferdestute bekannt ist, nachweisen. Die ER- und PR-Expression zeigt bei den zyklischen Maultierstuten ein sehr variables Bild (insbesondere Maultierstute 1), welches dem bei zyklierenden Pferdestuten während der physiologischen Decksaison nicht entspricht. Aufgrund der Diskrepanz zwischen klinischen Daten, Serumhormonkonzentrationen, endometrialer Funktionsmorphologie und Hormonrezeptorstatus bei den zyklischen Maultierstuten kann festgestellt werden, dass es bei den untersuchten Tieren nicht möglich ist, anhand der morphologisch-funktionellen Gegebenheiten am Endometrium auf den klinischen Zyklusstand zu schließen und vice versa. Die erhobenen Befunde sind in ihrer Gesamtheit am ehesten vergleichbar mit den Verhältnissen bei Pferdestuten in den Übergangszyklen zwischen Winteranöstrus und physiologischer Decksaison. Bei der Gruppe der azyklischen Maultiere (n=4) findet sich in allen untersuchten Endometriumbioptaten eine inaktive endometriale Funktionsmorphologie. Diese passt grundsätzlich zu dem klinischen Vorbericht der Azyklie mit inaktiven Ovarien und niedrigen, wenig schwankenden Serumöstradiol- und -progesteronkonzentrationen. In der Mehrzahl der Bioptate liegt eine Irregularität der glandulären Epithelien vor und in einigen Fällen treten hyperplastische und hypersekretorische Drüsenareale auf. Diese Befunde können, anlehnend an die Erkenntnisse bei der Pferdestute, als Kennzeichen einer endometrialen Fehldifferenzierung interpretiert werden (HÄFNER 1999, SCHOON et al. 1999b). Im Zusammenhang mit der schwachen endometrialen Proliferationsaktivität und der variablen ER- und PR-Expression lässt sich eine deutliche Ähnlichkeit zu den Befunden bei Pferdestuten im Winteranöstrus bzw. im Anöstrus während der physiologischen Decksaison finden (AUPPERLE et al. 2003, GOCKELN et al. 2006). Die möglichen Ursachen der endometrialen Fehldifferenzierung bei den Maultierstuten können im Rahmen der durchgeführten Untersuchungen nicht abschließend geklärt werden. Es ist jedoch denkbar, dass, analog zur Pferdestute, eine ovariell oder zentral bedingte Dysregulation und/oder eine Störung auf der Effektor/Rezeptor-Ebene vorliegen können. Die Hauptursache für die Infertilität bei Maultierstuten besteht in den chromosomal bedingten Störungen im Ablauf der Meiose der Keimzellen (WODSEDALEK 1916). Aufgrund der eigenen Untersuchungsergebnisse kann jedoch angenommen werden, dass die diagnostizierte endometriale Fehldifferenzierung, die variable Steroidhormonrezeptor-Expression sowie das Vorliegen degenerativer endometrialer Alterationen, z.B. der Endometrose, in Anlehnung an die Erkenntnisse bei der Pferdestute, zusätzliche Faktoren darstellen könnten, die zur Unfruchtbarkeit der Maultiere beitragen. Die Ergebnisse der vorliegenden Arbeit beruhen jedoch lediglich auf der Untersuchung von sechs Maultierstuten und sollten aus diesem Grund nicht unkritisch auf die Gesamtpopulation der Maultiere übertragen werden. Weitere Verlaufsuntersuchungen an einem größeren Tiergut in verschiedenen Altersgruppen inkl. Betrachtung der Hypophysenhormone FSH und LH könnten zur Aufklärung der Ursache sowie der Bedeutung der endometrialen Fehldifferenzierung für die Infertilität beim Maultier beitragen. / The aim of the present study was a detailed histomorphological characterization of the functional morphology and the evaluation of Ki-67 antigen, estrogen and progesterone receptor expression patterns in the endometrium of cyclic and non-cyclic mule mares, with respect to clinical and endocrinological data. The results were to be compared with corresponding observations in the endometrium of horse mares. For this purpose 97 endometrial biopsies from six clinically and gynaecologically healthy mule mares, aged between 13 and 23 years, were evaluated histologically and immunohistochemically. The biopsies were taken in intervals from two up to ten days over a period of four months (April to August 2004), in one mare over seven months (April to November 2004). Detailed information was given about clinical and gynaecological findings. The serum hormone concentrations for each day of biopsy sampling were determined (BARTMANN et al. 2008). According to the clinical record two of the animals show an erratic estrous cycle associated with the occurrence of corpora lutea and follicles on the ovaries during the whole period of investigation. Additionally, variable plasma estrogen and progesterone values are found. Four mules reveal acyclia including inactive ovaries and constant low serum hormone levels. In the animals of the cyclic group (n=2) the endometrial functional morphology varies to a great extend. In almost every investigated endometrial biopsy a marked endometrial maldifferentiation can be detected. It is classified according to common histomorphological criteria used for the endometrium of horse mares (HÄFNER 1999, SCHOON et al. 1999b). Endometrial maldifferentiation of the mules investigated occurs as irregular glandular or irregular and unequal differentiation and appeares in combination with a proliferative, secretory or inactive functional morphology. Mule 1 shows variable and mule 2 constant low endometrial Ki-67 expression patterns during the period of investigation. In contrast to horses, no characteristic, stage of cycle- or steroid hormone receptor expression-dependent proliferation activity can be observed with the mule mares of the present study. The expression of estrogen and progesterone receptors is highly variable in the animals of the cyclic group (especially in mule 1). These findings do not correspond to the ones in horse mares during the physiological breeding season. Due to the discrepancy between clinical data, serum hormone values, endometrial functional morphology and hormone receptor expression in the investigated mules, a conclusion from the clinical stage of cycle to morpho-functional conditions in the endometrium and vice versa seems to be impossible. The results of the present study are most likely comparable with findings in horse mares during the transitional period between winter anestrus and the physiological breeding season. The mules of the acyclic group (n=4) reveal an inactive endometrial functional morphology in all investigated endometrial biopsies. This finding basically corresponds to the clinical record of acyclia associated with inactive ovaries and rather constant serum estrogen and progesterone concentrations. In the case of these animals a majority of the investigated endometria show irregular glandular epithelia. In some areas hyperplastic and hypersecretory glands can be seen. According to observations in horse mares these findings can be interpreted as signs of endometrial maldifferentiation (HÄFNER 1999, SCHOON et al. 1999b). In connection with the weak proliferation activity and the variable ER- and PR-expression patterns seen in the acyclic mules, these results resemble findings in horses during winter anestrus or anestrus during the breeding season (AUPPERLE et al. 2003, GOCKELN et al. 2006). Within the present study the possible reasons for endometrial maldifferentiation in mules could not be finally evaluated. But it seems possible that, like in horse mares, an ovarian or central dysregulation and/or dysfunction regarding the level of receptors and effectors could play a certain role. Infertility in she-mules is mainly due to the chromosomal abnormalities during meiosis of germ cells (WODSEDALEK 1916). According to the findings in horse mares, the results of this investigation indicate that the diagnosed endometrial maldifferentiation, the variable expression of steroid hormone receptors and the presence of endometrial degenerative lesions (e.g. endometrosis) could be additional factors that possibly lead to sterility in mule mares, too. Due to the small number of individuals examined in this study, the results should not uncritically be applied to the whole population of mules. Further clinical, histological and immunohistochemical investigations, including the pituitary hormones FSH and LH as well as the use of a larger number of animals at different stages of age, could lead to a better understanding of the causes of endometrial maldifferentiation and its relevance for the infertility in mules.

A isquemia cerebral é uma enfermidade grave, de incidência e prevalência global, considerada uma das principais causas de morte e incapacidade no mundo. Ela é decorrente da hipoperfusão ou interrupção sanguínea em uma determinada região encefálica e a falta de oxigênio e glicose leva a falha no metabolismo energético de neurônios e células gliais. A morte destas células libera grande quantidade de glutamato, o principal neurotransmissor excitatório, o que desencadeia a cascata isquêmica, formada por estresse oxidativo, excitotoxicidade e processo inflamatório. Estes insultos causam danos teciduais graves que comprometem, principalmente, o sistema sensorimotor, memória, cognição e as emoções. A guanosina, um derivado da guanina, tem mostrado papel neuroprotetor ao sistema nervoso central, agindo no sistema glutamatérgico. Esses efeitos têm sido demonstrados em ratos machos, mas não em fêmeas, por isso, o objetivo desta tese foi investigar através de testes comportamentais, imunoistoquímico e histológico, a ação da guanosina em ratos Wistar machos e fêmeas induzidos cirurgicamente a um modelo de isquemia focal permanente do córtex cerebral parietal, levando em consideração o ciclo estral das fêmeas imediatamente antes da indução isquêmica. Este trabalho foi aprovado pela Comissão de Ética no Uso de Animais da UFRGS (Nº 29396). Após a verificação do ciclo estral, fêmeas e machos foram anestesiados, posicionados em aparelho estereotáxico e a isquemia focal cortical foi induzida por termocoagulação dos vasos piais. Ao término da cirurgia, os animais foram tratados com solução salina 0,9% ou guanosina 60 mg/kg, ambos por via intraperitoneal, em 4 doses (0, 1, 3 e 6 horas após a indução isquêmica). No teste do cilindro, as fêmeas apresentaram significativamente melhor recuperação sensorimotora a longo prazo comparadas aos machos. No teste do campo aberto, memória de habituação a longo prazo e locomoção em fêmeas foram prejudicadas pela isquemia e os machos não apresentaram prejuízo. No teste do labirinto em cruz elevado não houve diferença significativa entre os grupos. No teste claro/escuro, as fêmeas isquêmicas demonstraram comportamento ansiolítico comparadas aos machos e às fêmeas naïves. A imunoistoquímica de astrócitos e a mensuração do volume de lesão não apresentaram diferença estatística entre os grupos. Com este trabalho demonstrou-se a importância da realização de pesquisas em ambos os gêneros, considerando o ciclo estral, principalmente para enfermidades complexas como a isquemia cerebral. O tratamento com a guanosina é promissor, principalmente na recuperação motora em fêmeas, entretanto, estudos futuros são necessários para melhor compreendimento dos mecanismos envolvidos. / Ischemic stroke is a serious disease of global incidence and prevalence, considered a leading cause of death and disability worldwide. It is due to hypoperfusion or blood disruption in a brain region and the lack of oxygen and glucose leads to failure in energetic metabolism of neurons and glial cells. The death of these cells releases large amount of glutamate, the main excitatory neurotransmitter, which triggers the ischemic cascade, formed by oxidative stress, excitotoxicity and inflammatory processes. These insults cause severe tissue damage that mainly compromise the sensorimotor system, memory, cognition and emotions. Guanosine, a guanine derivative, has shown a neuroprotective role in the central nervous system, acting on the glutamatergic system. These effects have been demonstrated in male rats, but not in females, so the aim of this thesis was to investigate through behavioral, immunohistochemical and histological tests, the action of guanosine in male and female Wistar rats surgically induced to a model of focal permanent cerebral ischemia in the motor cortex, considering the female estrous cycle immediately before the ischemic induction. This work was approved by the UFRGS Ethical Committee on the Use of Animals (No. 29396). After estrous cycle verification, females and males were anesthetized, positioned in a stereotaxic apparatus, and cortical focal ischemia was induced by thermocoagulation of the pial vessels. At the end of the surgery, the animals were treated with 0.9% saline or guanosine 60 mg/kg, both intraperitoneally, in 4 doses (0, 1, 3 and 6 hours after ischemic induction). In the cylinder test, females presented significant long-term sensorimotor recovery compared to males. In the open field test, long-term memory habituation and locomotion/exploratory activity in females were impaired by ischemia and males showed no impairment. In the elevated plus maze test, there was no significant difference between groups. In the light/dark test, the ischemic females showed anxiolytic-like behavior compared to naïve males and females. The immunohistochemistry of astrocytes and the measurement of lesion volume did not show any statistical difference between groups. With this work it was demonstrated the importance of conducting researches in both genders, considering the estrous cycle, mainly for complex diseases such as cerebral ischemia. The treatment with guanosine is promising, especially in motor recovery in females; however, future studies are necessary to better understand the mechanisms involved.

L'utérus est un organe hautement dynamique, qui subit des changements morphologiques durant le cycle œstral sous l'influence des hormones sexuelles. Ces adaptations sont essentielles à l’homéostasie utérine et résultent d’une régulation minutieuse de la prolifération, de la différenciation et de la survie cellulaire par de nombreuses voies de signalisation. Nous avons émis l'hypothèse que la voie Hippo est impliquée dans l'homéostasie utérine chez l’espèce bovine, et nous avons tenté de déterminer les variations de ses constituants. Des biopsies de l'endomètre et des échantillons de sang ont été obtenus aux jours 0, 3 et 10 du cycle chez 14 vaches. Les concentrations sanguines de progestérone et d’œstrogène ont confirmé la progression du cycle œstral chez cinq vaches. Des analyses immunohistochimiques, Western Blot et RT-PCR ont été réalisées sur les biopsies afin d’évaluer la variation des composantes de la voie Hippo et de ses gènes cibles. Nous avons démontré la présence de YAP et de TAZ dans l'endomètre bovin à tous les moments du cycle œstral. Elles présentaient une expression cytoplasmique dans l'épithélium glandulaire, et présentaient une expression nucléaire dans le stroma à J0 et J10 et dans l'épithélium luminal à J3. Nous avons également noté une tendance à l'augmentation des concentrations de la forme phosphorylée de YAP au jour 3 du cycle œstral par rapport aux jours 0 et 10. Cette augmentation a été corrélée à une diminution de l'expression du gène CTGF au jour 3 par rapport aux autres jours du cycle œstral. En conclusion, nous avons démontré que la voie Hippo est exprimée différentiellement dans l'utérus bovin au cours du cycle œstral. / The uterus is a highly dynamic organ that changes morphologically under the influence of sexual hormones during the estrous cycle. These adaptations are essential for the proper function of the uterus and are due to the careful regulation of cell proliferation, differentiation, and apoptosis by numerous signaling pathways. We hypothesized that the Hippo pathway is implicated in uterine homeostasis in cattle, and we aimed to determine the variation of its canonical constituents during the bovine cycle. Endometrial biopsies and blood samples were obtained on days 0, 3, and 10 of the cycle in 14 cows. Progesterone and estrogens blood concentrations were measured and confirmed the status of the oestrus cycle in five cows. Immunohistochemistry, western blot, and RT-PCR were then performed on their endometrial biopsies to evaluate the Hippo components and targeted genes. Our results showed the presence of YAP and TAZ in the bovine endometrium at all time point. Both proteins displayed cytoplasmic expression in the glandular epithelium while exhibiting nuclear expression in the stroma on D0 and D10 and in the luminal epithelium on D3. We also noted a tendency for an increased amount of phosphorylated YAP during day 3 of the estrous cycle compared to days 0 and 10. It was correlated with a decreased expression of CTGF (Hippo target gene) on day 3 compared to the other day of the cycle. In conclusion, we can confirm that Hippo is differentially expressed in the bovine uterus during the estrous cycle.