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.

Orientador: Guilherme de Paula Nogueira / Banca: José Luis Moraes Vasconcelos / Banca: Ciro Moraes Barros / Resumo: Seis novilhas Nelore tiveram seus ciclos estrais acompanhados durante diferentes estações do ano (outono n=11; inverno n=8; primavera n=9; verão n=9) com exames ultrassonográficos diários para contar e mensurar folículos ≥3mm. Amostras de sangue foram colhidas a cada 12h para hormônio luteinizante (LH) e progesterona (P4), e a cada 3h do estro até a ovulação para caracterizar o pico de LH. Cinco novilhas ovariectomizadas receberam 17β-estradiol (2μg/kg/p.v.) em cada estação, e amostras de sangue foram colhidas depois disso a cada 3h para quantificação de LH. A diferença percentual mensal ( %) do peso não variou entre as estações. A concentração média de P4 no ciclo estral foi maior (p=0,001) e o número de folículos menor (p=0,001) durante o outono (2,5±0,2ng/mL; 7,8±0,1) e verão (2,9±0,3ng/mL; 6,8±0,2) comparado com o inverno (1,4±0,2ng/mL; 9,6±0,3) e primavera (1,6±0,2ng/mL; 9,7±0,3). Durante o inverno houve mais ciclos estrais com três (5 de 8) e durante o verão somente ciclos com duas ondas foliculares (p=0,009). Como a secreção de LH não variou, apesar da variação sazonal na concentração de P4, e como houve correlação negativa entre os valores máximos de P4 e a variação percentual do fotoperíodo (p=0,0056; r = -0,4465), uma variação sazonal na sensibilidade das células luteínicas ao LH precisa ser avaliada. Nas novilhas ovariectomizadas, a concentração circanual de LH sem o estímulo de estradiol foi significante (p=0,0214). A resposta de LH ao tratamento de estradiol foi menor no verão (0,8±0,2ng/mL vs 1,3±0,5ng/mL). Nós supomos que existe variação sazonal na sensibilidade hipotalâmica ao estradiol. / Abstract: Six Nelore heifers had their estrous cycle followed during different seasons of the year (autumn n=11; winter n=8; spring n=9 and summer n=9) with daily ultrasonographic exams to count and measure follicles ≥3mm. Blood was collected every 12h for luteinizing hormone (LH) and progesterone (P4), and every 3h from estrus until ovulation to characterize the LH peak. Five ovariectomized heifers were injected with 17β-estradiol (2μg/kg/LW) every season and blood samples were collected thereafter at 3h intervals for LH quantification. The monthly body weight percentile difference ( %) did not vary between seasons. Average P4 concentration for the cycle was higher (p=0.001) and follicle number lower during autumn (2.5±.2ng/ml; 7.8±.1) and summer (2.9±.3ng/ml; 6.8±.2) (p=0.001) compared to winter (1.4±.2ng/ml; 9.6±.3) and spring (1.6±.2ng/ml; 9.7±.3). During winter there were more estrous cycles with three follicle waves (5 out of 8) and during summer only cycles with two follicular waves (p=0.009). As LH secretion did not vary despite seasonal variation in P4 concentration and as there was a negative correlation between higher P4 values and daily percentile variation of photoperiod ( %, p=0.0056; r= -0.4465), a seasonal variation in luteal cell sensitivity to LH needs to be evaluated. In the ovariectomized Nelore heifers, the LH circanual concentration without estradiol stimulus was significant (p=0.0214). The LH response to estradiol treatment was lower in summer (0.8±.2ng/ml vs 1.3±.5ng/ml). We hypothesize there exists seasonal variation in hypothalamic sensitivity to estradiol. / Mestre

A matriz extracelular (MEC) dos tecidos uterinos é altamente remodelada na gestação de camundongos. Os objetivos deste estudo foram avaliar a influência dos hormônios ovarianos estrógeno (E2) e progesterona (P4) sobre a estrutura dos tecidos uterinos de camundongo e a deposição dos proteoglicanos decorim, biglicam, fibromodulim, lumicam, perlecam e versicam nestes tecidos. Para isto, utilizamos um modelo de castração e reposição hormonal, e o ciclo estral como parâmetro fisiológico. Verificamos que, como na gestação, durante o ciclo estral ocorre intensa remodelação na estrutura e na MEC dos tecidos uterinos. Verificamos ainda que a resposta aos hormônios ovarianos é: compartimento-específica; hormônio-específica e molécula-específica. Notável foi a modulação do versicam pelos hormônios ovarianos. P4 induz a deposição de versicam no estroma, enquanto o miométrio responde apenas a E2. A modulação dos proteoglicanos pelos hormônios ovarianos mostra a relevância destas moléculas para a composição de um ambiente uterino propício para o desenvolvimento do embrião. / The extracellular matrix (ECM) of the mouse uterine tissues is highly remodeled during pregnancy. The aim of this study was to demonstrate the influence of estrogen (E2) and progesterone (P4) on the uterine structure and on the deposition of the proteoglycans decorin, biglycan, fibromodulin, lumican, perlecan and versican in these tissues. For that purpose, we used a model of castration e hormone replacement as main strategy, and the estrous cycle as physiological parameter. We verified that, as in pregnancy, during the estrous cycle an intense remodeling occurs on the structure and the ECM of the uterine tissues. We also showed that the response to the ovarian hormones is: compartment-; hormone- and molecule- specific. Noteworthy was the modulation of versican by the hormones: P4 induces the deposition of versican in the stroma, whereas the myometrium responds only to E2. The modulation of proteoglycans by the ovarian hormones indicates the relevance of these molecules for the composition of a proper microenvironment for embryo development.

Os ratos-de-espinho são abundantes nas florestas úmidas Neotropicais. Trinomys yonenagae (ROCHA, 1995) constitui uma exceção entre as espécies do gênero por habitar o campo de dunas fixas da Caatinga, ser colonial e fossorial. Tanto o ambiente seco, como a socialidade e o hábito fossorial podem alterar o padrão reprodutivo das espécies. Diante do fato desses roedores serem coloniais, por exemplo, espera-se a influência de co-específicos (fêmeas ou machos) no ciclo estral. Estudos-piloto na natureza e em cativeiro mostraram que a fase de diestro pode ocorrer com a vagina aberta, um fato que merece ser mais investigado, já que se considera que a abertura vaginal ocorra somente durante a transição da fase proestro-estro. O objetivo deste trabalho é, portanto, o de estudar aspectos dos ritmos reprodutivos infradianos (ciclo estral e gestação) e circadiano (temperatura corporal) em rabos-de-facho, mantidos em cativeiro, alojados individualmente, e pareados com fêmeas e com machos. Nesses grupos são caracterizados os esfregaços e o estado de abertura da vagina, buscando conhecer aspectos da fisiologia reprodutiva, tais como, a idade de maturidade sexual (cap. 1), a relação entre a fase do ciclo estral e o estado vaginal (cap. 2), e as características da citologia esfoliativa durante a gravidez (cap. 3). Os dados da citologia são correlacionados com o padrão comportamental durante o acasalamento, caracterizando a existência de estro pós-parto (cap. 3). Analisa-se ainda a distribuição de todos os nascimentos ocorridos ao longo de dez anos. O ritmo de temperatura corporal das fêmeas é aferido a partir de acompanhamento da temperatura retal com uso de termômetro digital durante 72h (cap. 4). Embora de maneira geral Trinomys yonenagae siga os padrões reprodutivos de outros membros da família Echimyidae, tais como, gestação longa (90d), ninhadas pequenas (média de 2 filhotes) e maturidade sexual tardia (ao redor dos 90d) é possível identificar a influência de co-específicos (dominância entre fêmeas) no ciclo estral. Este ciclo parece ser espontâneo, do tipo poliéstrico, com fases muito curtas de proestro e estro (< 24h). A condição vaginal não é um bom indicativo de atividade reprodutiva (ocorre abertura no diestro e na gravidez); a presença do macho não garante fecundação; também se confirma a ocorrência de estro pós-parto, igualmente breve. O comportamento de acasalamento é elaborado, o que é esperado de espécies com estro curto ou de vida social e subterrânea. A distribuição dos nascimentos não é sazonal, confirmando dados de campo. Quanto ao ritmo de temperatura corporal, ele é circadiano com picos noturnos e com relação positiva de fase com o ritmo de atividade/repouso, o que segue o padrão geral de roedores / Spiny rats are among the most numerous group of mammals in Neotropical rainforests. Trinomys yonenagae (ROCHA, 1995) is an exception because it is colonial, fossorial and inhabits the semiarid Caatinga. The dry environment and the underground and colonial life styles are known to affect reproductive patterns. So it is expected, for example, that social life may influence the estrous cycle especially through the presence of conspecific (females and males). It is considered that vaginal opening occurs only during the estrus. Previous field studies or in captivity have shown the unexpected occurrence of vaginal opening and diestrus, a fact that deserves further investigation. The aim of this work is verify the reproductive infradian rhythms (estrous cycle and pregnancy) and the circadian body temperature rhythm, in T. yonenagae kept individually or in pairs (male-female and female-female) in captivity. In these groups vaginal smears and the presence of the vaginal membrane is studied aiming to characterize aspects of its reproductive life, such as, the age of sexual maturity (chapter 1), the relationship between the vaginal opening and the phase of the estrous cycle (chapter 2), and the smear cytology during pregnancy (chapter 3). The cytological data are correlated with the mating behavioral, characterizing the post-partum estrus (chapter 3). All birth that occurrence during 10 years are analyzed. The body temperature rhythm is measured during 72h by rectal temperature with a digital thermometer (chapter 4). Although Trinomys yonenagae follows the reproductive patterns of other members of Echimyidae, such as prolonged gestation( 90d), small sized litter (mean of 2) and delayed sexual maturity (around 90d), it is possible to identify the influence of conspecifics (female dominance) upon the estrous cycle. This cycle seems to be spontaneous, poliestric, having very short proestrus and estrus phases (< 24h). The vaginal condition is not a good indicative of the reproductive status (vaginal opening may occur during the diestrus phase and the pregnancy); the presence of males does not guarantee pregnancy. We also confirm the occurrence of post-partum estrus, which is also short-lived. The mating behavior is complex, which is expected in species were the duration of estrus is short, or in species of social and subterranean life. The birth distribution is non seasonal, which is in agreement with field data. The body temperature rhythmicity is circadian with peaks at night; and shows a positive phase relationship with the rest/activity rhythm, both cycles follow the pattern known for other nocturnal rodents

Avaliamos o efeito do treinamento físico intenso e da restrição alimentar no eixo reprodutor e na expressão do RNAm do MCH em ratas fêmeas. Utilizamos ratas fêmeas sedentárias alimentadas (SA), sedentárias com restrição alimentar, treinadas alimentadas (TA) e treinadas com restrição alimentar. O grupo TR entrou em anestro a partir da quarta semana de experimento. Os grupos restritos apresentaram redução de peso corporal, ovários, úteros e tecido adiposo branco, comparados aos alimentados. Houve redução sérica de leptina nos grupos SR e TR. Os grupos TA, SR e TR apresentaram cistos ovarianos. A expressão do RNAm do ppMCH na área incerto-hipotalâmica (IHy) foi menor nos animais TR, SR e nos animais alimentados em proestro, e na área hipotalâmica lateral (LHA) foi maior nos animais em restrição alimentar e nos animais alimentados em proestro. Conclui-se que a leptina exerce forte influencia no eixo reprodutor feminino, tal ação parece ter envolvimento do MCH, sobretudo o proveniente da IHy, uma vez que o MCH da LHA tem relação com o estímulo a busca de alimento. / We evaluated the effect of intense physical training and food restriction on the reproductive axis and MCH mRNA expression in female rats. We used female rats fed sedentary (SF), food-restricted sedentary, trained fed (TF) and trained with food restriction. The group entered anestrus TR from the fourth week of the experiment. The restricted groups had reduced body weight, ovary, uterus and white adipose tissue, compared to the fed. There was a reduction of serum leptin in the SR and TR groups. The TF groups, SR and TR had ovarian cysts. The expression of mRNA ppMCH incerto-hypothalamic area (IHy) was lower in TR animals, SR and animals fed on proestrus, and the lateral hypothalamic area (LHA) were higher in animals on restricted food and animal feeding in proestrus. It is concluded that leptin exerts a strong influence on female reproductive axis, such action appears to have involvement of MCH, especially from the IHy, since the MCH of the LHA is related to the motivational behavior associated to the search for food.

Avaliamos o efeito do treinamento físico intenso e da restrição alimentar no eixo reprodutor e na expressão do RNAm do MCH em ratas fêmeas. Utilizamos ratas fêmeas sedentárias alimentadas (SA), sedentárias com restrição alimentar, treinadas alimentadas (TA) e treinadas com restrição alimentar. O grupo TR entrou em anestro a partir da quarta semana de experimento. Os grupos restritos apresentaram redução de peso corporal, ovários, úteros e tecido adiposo branco, comparados aos alimentados. Houve redução sérica de leptina nos grupos SR e TR. Os grupos TA, SR e TR apresentaram cistos ovarianos. A expressão do RNAm do ppMCH na área incerto-hipotalâmica (IHy) foi menor nos animais TR, SR e nos animais alimentados em proestro, e na área hipotalâmica lateral (LHA) foi maior nos animais em restrição alimentar e nos animais alimentados em proestro. Conclui-se que a leptina exerce forte influencia no eixo reprodutor feminino, tal ação parece ter envolvimento do MCH, sobretudo o proveniente da IHy, uma vez que o MCH da LHA tem relação com o estímulo a busca de alimento. / We evaluated the effect of intense physical training and food restriction on the reproductive axis and MCH mRNA expression in female rats. We used female rats fed sedentary (SF), food-restricted sedentary, trained fed (TF) and trained with food restriction. The group entered anestrus TR from the fourth week of the experiment. The restricted groups had reduced body weight, ovary, uterus and white adipose tissue, compared to the fed. There was a reduction of serum leptin in the SR and TR groups. The TF groups, SR and TR had ovarian cysts. The expression of mRNA ppMCH incerto-hypothalamic area (IHy) was lower in TR animals, SR and animals fed on proestrus, and the lateral hypothalamic area (LHA) were higher in animals on restricted food and animal feeding in proestrus. It is concluded that leptin exerts a strong influence on female reproductive axis, such action appears to have involvement of MCH, especially from the IHy, since the MCH of the LHA is related to the motivational behavior associated to the search for food.