Made available in DSpace on 2014-07-29T15:07:30Z (GMT). No. of bitstreams: 1 Ubirajara_Bilego.pdf: 525711 bytes, checksum: 92b1baad77eca0e7758c3f7bfe166098 (MD5) Previous issue date: 2009-02-26 / The aim of this study was to describe the physiological responses to the Ovsynch protocol in Girolando heifers breeding in extensive model on Centro-Oeste of Brazil region and to determine relationships with environments factors in two stations, dry and rain. This responses were describe through the characterization of ovarian structures how: (NTL) total number of follicles; (DFOLIPR) mean of follicular diameter on protocol s begin; (DFOLOV) ovulatory diameter follicles; (DCL) corpus luteum ovulatory diameter; (SCL) ovulatory corpus luteum brightness score; pregnancy rates and ovulation rate on day 9 obtained in two year stations. Were utilized 40 heifers 18-24 months age, split in two groups of 20 animals being G1 group of dry station and G2 group of rain station. During the evaluations were found differences for the total number of follicles 8,07±0,25 e 8,83±0,26 in two stations applied. The mean of follicular diameter on protocol s begin was not show differences, but the measure of the ovulatory follicle diameter, have differences for the year stations (P<0,01), being the diameter for the dry station 11,88 ± 0,4mm whereas the rain station was 10,13 ± 0,36mm. Have in this study, higher frequency of follicles that ovulate with 11 to 12 mm on dry station and 10 to 11mm on rain station. The mean of the corpus luteum diameter on the present study not differ among the stations (P>0,05), with10,46±0,41mm for the dry station and 10,49±0,34mm for the rain station, being the higher frequency of CL 11 to 12mm in both stages. The CL brightness score not have differ(P>0,05), being 2,6 ± 0,16 for the dry and 2,31 ± 0,11 for the rain. The pregnancy rate have differences on two stages (P<0,01) with values 45% and 11% on station dry and rain respectively. The THI values differ among the stations dry and rain (P<0,01). The ovulation rate on D9 do not differ (P>0,05) although to have numeric differences. It was possible to conclude that in two stations evaluated, dry and rain, have differences on follicular morfometry and pregnancy rates. / Objetivou-se com este estudo descrever a resposta fisiológica do protocolo Ovsynch em novilhas Girolando, criadas extensivamente na região Centro-Oeste do Brasil em duas estações do ano, seca e águas. Tais respostas foram descritas através da caracterização das estruturas ovarianas como: (NTF) Número total de folículos; (DFOLIPR) Diâmetro folicular médio no início do protocolo; (DFOLOV) Diâmetro do folículo ovulatório; (DCL) Diâmetro do corpo lúteo; (SCL) Escore de ecogenicidade do corpo lúteo; Taxa de gestação e Taxa de ovulação no D9 obtidas em duas estações do ano. Foram utilizadas 40 novilhas girolando com idades entre 18-24 meses, divididas em dois grupos de 20 animais, sendo (G1) de estação seca e (G2) grupo de estação de águas. Durante as avaliações foram encontradas diferenças quanto ao número total de folículos 8,07±0,25 e 8,83±0,26 nas duas estações avaliadas. O diâmetro folicular médio no início do protocolo não mostrou diferenças, porém, houve diferença entre as estações do ano (P< 0,01), quanto ao diâmetro folicular ovulatório sendo o diâmetro médio para a estação seca 11,88 ± 0,4mm enquanto para a estação das águas foi 10,13 ± 0,36mm. Houve nesse estudo maior freqüência de folículos que ovularam em torno de 11 a 12mm na estação seca e 10 a 11mm na estação das águas. O diâmetro médio do corpo lúteo no presente estudo não diferiu entre as estações (P>0,05), com 10,46±0,41mm para a estação seca e 10,49±0,34mm, com maior freqüência de CLs de 11 a 12mm em ambas as estações. Os escores de corpo lúteo também não mostraram diferenças (P>0,05) sendo 2,6 ± 0,16 para seca e 2,31 ± 0,11 para águas. A taxa de gestação também mostrou diferença entre as estações com valores de 45% e 11% nas estações seca e águas respectivamente. Taxa de ovulação no D9 não diferiu (P>0,05) entre as estações apesar de ser numericamente diferente entre os tratamentos. Os valores de THI diferiram entre as estações seca e águas (P<0,01). Foi possível concluir que as duas estações avaliadas, seca e águas, mostraram diferenças quanto morfometria folicular e taxas de gestação.

Objetivo: avaliar marcadores de reserva ovariana e a presença de anticorpo anti-corpo lúteo (anti-CoL) em pacientes com lúpus eritematoso sistêmico (LES) de início na infância. Métodos: A presença do anti-CoL foi avaliada através de immunoblot em cinquenta e sete mulheres com LES e 21 controles saudáveis. A reserva ovariana foi estimada através das dosagens do hormônio folículo estimulante (FSH), hormônio luteinizante (LH), estradiol, hormônio anti- Mülleriano (AMH) e da contagem de folículos antrais (CFA). Foram também avaliados dados demográficos, alterações menstruais, atividade da doença, dano cumulativo e tratamento. Resultados: a mediana da idade atual foi similar nos pacientes com LES em relação aos controles (27,7 vs. 27,7 anos, p=0,414). A mediana do AMH (1,1 vs. 1,5ng/mL, p=0,037) da CFA (6 vs. 16 p<0,001) forma significantemente menores nos pacientes com LES quando comparados aos controles, porém sem alterações menstruais significantes. A presença do anti-CoL foi observada apenas nos pacientes com LES (16% vs. 0%, p=0,103) e não foi relacionada com dados demográficos, parâmetros de reserva ovariana, atividade da doença, dano cumulativo ou tratamento. Avaliação dos pacientes tratados com ciclofosfamida mostrou níveis elevados de FSH quando comparados com os pacientes que não receberam ciclofosfamida e com controles (8,8 vs. 5,7 vs. 5,6IU/L, p=0,032) e níveis menores de AMH e CFA (0,4 vs. 1,5 vs. 1,5ng/mL, p=0,004; 4,0 vs. 6,5 vs. 16IU/L, p=0,001; respectivamente). Dezenove pacientes foram tratados com metotrexate sem histórico de uso de ciclofosfamida sendo evidenciada uma correlação negativa entre a dose cumulativa de metotrexate e os níveis de AMH (p=0,027, r=-0,507). Conclusões: este estudo identificou que altas doses ciclofosfamida e metotrexato são causas relevantes de disfunção ovariana subclínica durante a idade reprodutiva em mulheres com LES de início na infância / Objective: To assess ovarian reserve markers and anti-corpus luteum antibodies (anti-CoL) in systemic lupus erythematosus (SLE) patients with onset before adulthood. Methods: Fifty-seven SLE female patients and 21 healthy controls were evaluated for anti-CoL by immunoblot. Ovarian reserve was assessed by: follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, anti-Müllerian hormone (AMH) and antral follicle count (AFC). Demographic data, menstrual abnormalities, disease activity, damage and treatment were also analyzed. Results: The median of current age was similar in SLE patients and controls (27.7. vs. 27.7 years, p=0.414). The median of AMH (1.1 vs. 1.5ng/mL, p=0.037) and AFC (6 vs. 16, p<0.001) were significantly reduced in SLE patients versus controls without significant menstrual abnormalities. Anti-CoL was solely observed in SLE patients (16% vs. 0%, p=0.103) and not associated with demographic data, ovarian reserve parameters, disease activity/damage and treatment. Further evaluation of patients treated with cyclophosphamide revealed a higher median of FSH levels compared to SLE patients not treated with cyclophosphamide and controls (8.8 vs. 5.7 vs. 5.6IU/L, p=0.032) and a lower median AMH levels and AFC (0.4 vs. 1.5 vs. 1.5ng/mL, p=0.004; 4.0 vs. 6.5 vs. 16IU/L, p=0.001; respectively). Nineteen patients were treated with methotrexate without cyclophosphamide use, and a negative correlation was observed between cumulative methotrexate dose and AMH levels (p=0.027, r=-0.507). Conclusions: The present study identifies high doses of cyclophosphamide and methotrexate as relevant causes of subclinical ovarian dysfunction during reproductive ages in SLE patients with onset before adulthood

Der Gelbkörper (corpus luteum, CL) ist eine transitorische Drüse, die im Ovar nach der Ovulation gebildet wird und durch ihre Progesteron-(P4)-Produktion die Trächtigkeit unterstützt. Bei allen bisher untersuchten Säugetieren endet die sekretorische Aktivität der CL mit dem Ende der Trächtigkeit oder Laktation, danach werden die CL abgebaut. Der Zyklus von Katzenartigen, wie etwa der Hauskatze, folgt dem gleichen Muster. Anders verläuft der Zyklus bei Luchsen. Beim Eurasischen Luchs (Lynx lynx) und beim Pardelluchs (Lynx pardinus) persistieren die CL nach der Geburt für mehr als zwei Jahre und sezernieren weiter P4. Die vorgestellte Arbeit sollte die Funktion persistierender (per) CL im Luchs untersuchen und die Fortpflanzung von Katzen weiter aufklären. Dazu wurden grundlegende histologische und hormonelle Aspekte der Lutealphase analysiert und der Einfluss des apoptotischen Systems sowie die Rezeptivität gegenüber Steroiden bei der Regulation der CL-Funktion betrachtet. Die CL von Hauskatzen und Luchsen wurden histomorphologisch unterteilt. In allen Proben wurden intraluteales P4 und Östrogene bestimmt. Weiterhin wurde die mRNA- und, wenn möglich, die Proteinexpression der proapoptotischen Faktoren BAX, Caspase-3, FAS, Tumor necrosis factor (TNF), TNF Rezeptor 1 (TNFRSFA1) und der Überlebensfaktoren (BCL2, TNFRSFB1), sowie des Progesteronrezeptors (PGR), der PGR-Membrankomponente (PGRMC) 1 und 2, des Östrogenrezeptors (ESR) 1 und 2, des G-Protein-gekoppelten Östrogenrezeptors 1 (GPER1) und des Androgenrezeptors (AR) gemessen. Die Ergebnisse weisen darauf hin, dass die Lutealphase der Hauskatze durch FAS, Caspase-3 und die TNF Rezeptoren 1 und 2 reguliert sein könnte. Steroide könnten über ihre Rezeptoren PGR, PGRMC1 und PGRMC2, ESR1 und AR wirken. Die physiologische Persistenz der CL beim Luchs könnte über BCL2, FAS, TNFRSFB1, PGRMC1, PGRMC2, ESR1, GPER1 und AR vermittelt werden. / Corpus luteum (CL) is a transitory gland which forms in the ovary after ovulation and supports the pregnancy with its production of progesterone (P4). In all mammals studied so far, the CL loses its secretory activity after pregnancy and regresses from the ovary. The feline luteal cycle follows the same pattern, and CL of the domestic cat functionally and structurally regress after lactation. However, the story is different for the lynx. In the Eurasian (Lynx lynx) and Iberian (Lynx pardinus) lynx, CL persist after parturition, weaning and for up to two years, still retaining their ability to secrete P4. Current work was initiated to understand the control of unusual persistent (per) CL in lynx and to learn more about feline reproduction in general. For this, studies on the basic histological and endocrinological aspects of the feline luteal phase, as well as potential involvement of systems of apoptosis and steroid receptivity in the CL regulation were performed. Collected CL from domestic cats and lynx were classified based on their histomorphology. In all samples, intraluteal P4 and estrogens were measured. Moreover, mRNA and where possible protein levels were determined for pro-apoptotic BAX, caspase-3, FAS, tumor necrosis factor (TNF), TNF receptor 1 (TNFRSFA1), pro-survival BCL2, TNFRSFB1, and for progesterone receptor (PGR), PGR membrane components (PGRMC) 1 and 2, estrogen receptors (ESR) 1 and 2, G protein-coupled estrogen receptor 1 (GPER1) and androgen receptor (AR). The results suggest that the luteal phase of the domestic cat is potentially regulated by caspase-3, FAS, TNFRSF1A, TNFRSF1B, and by actions of steroids via PGR, PGRMC1, PGRMC2, ESR1 and AR. Physiological persistence of Iberian lynx CL might be mediated by BCL2, FAS, TNFRSFB1, PGRMC1, PGRMC2, ESR1, GPER1 and AR. Current work indicates profound differences between the CL function and regulation in domestic cats and lynx, and promotes a highly species-specific approach in reproduction studies.

Objetivo: avaliar marcadores de reserva ovariana e a presença de anticorpo anti-corpo lúteo (anti-CoL) em pacientes com lúpus eritematoso sistêmico (LES) de início na infância. Métodos: A presença do anti-CoL foi avaliada através de immunoblot em cinquenta e sete mulheres com LES e 21 controles saudáveis. A reserva ovariana foi estimada através das dosagens do hormônio folículo estimulante (FSH), hormônio luteinizante (LH), estradiol, hormônio anti- Mülleriano (AMH) e da contagem de folículos antrais (CFA). Foram também avaliados dados demográficos, alterações menstruais, atividade da doença, dano cumulativo e tratamento. Resultados: a mediana da idade atual foi similar nos pacientes com LES em relação aos controles (27,7 vs. 27,7 anos, p=0,414). A mediana do AMH (1,1 vs. 1,5ng/mL, p=0,037) da CFA (6 vs. 16 p<0,001) forma significantemente menores nos pacientes com LES quando comparados aos controles, porém sem alterações menstruais significantes. A presença do anti-CoL foi observada apenas nos pacientes com LES (16% vs. 0%, p=0,103) e não foi relacionada com dados demográficos, parâmetros de reserva ovariana, atividade da doença, dano cumulativo ou tratamento. Avaliação dos pacientes tratados com ciclofosfamida mostrou níveis elevados de FSH quando comparados com os pacientes que não receberam ciclofosfamida e com controles (8,8 vs. 5,7 vs. 5,6IU/L, p=0,032) e níveis menores de AMH e CFA (0,4 vs. 1,5 vs. 1,5ng/mL, p=0,004; 4,0 vs. 6,5 vs. 16IU/L, p=0,001; respectivamente). Dezenove pacientes foram tratados com metotrexate sem histórico de uso de ciclofosfamida sendo evidenciada uma correlação negativa entre a dose cumulativa de metotrexate e os níveis de AMH (p=0,027, r=-0,507). Conclusões: este estudo identificou que altas doses ciclofosfamida e metotrexato são causas relevantes de disfunção ovariana subclínica durante a idade reprodutiva em mulheres com LES de início na infância / Objective: To assess ovarian reserve markers and anti-corpus luteum antibodies (anti-CoL) in systemic lupus erythematosus (SLE) patients with onset before adulthood. Methods: Fifty-seven SLE female patients and 21 healthy controls were evaluated for anti-CoL by immunoblot. Ovarian reserve was assessed by: follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, anti-Müllerian hormone (AMH) and antral follicle count (AFC). Demographic data, menstrual abnormalities, disease activity, damage and treatment were also analyzed. Results: The median of current age was similar in SLE patients and controls (27.7. vs. 27.7 years, p=0.414). The median of AMH (1.1 vs. 1.5ng/mL, p=0.037) and AFC (6 vs. 16, p<0.001) were significantly reduced in SLE patients versus controls without significant menstrual abnormalities. Anti-CoL was solely observed in SLE patients (16% vs. 0%, p=0.103) and not associated with demographic data, ovarian reserve parameters, disease activity/damage and treatment. Further evaluation of patients treated with cyclophosphamide revealed a higher median of FSH levels compared to SLE patients not treated with cyclophosphamide and controls (8.8 vs. 5.7 vs. 5.6IU/L, p=0.032) and a lower median AMH levels and AFC (0.4 vs. 1.5 vs. 1.5ng/mL, p=0.004; 4.0 vs. 6.5 vs. 16IU/L, p=0.001; respectively). Nineteen patients were treated with methotrexate without cyclophosphamide use, and a negative correlation was observed between cumulative methotrexate dose and AMH levels (p=0.027, r=-0.507). Conclusions: The present study identifies high doses of cyclophosphamide and methotrexate as relevant causes of subclinical ovarian dysfunction during reproductive ages in SLE patients with onset before adulthood

The control of reproductive cycles in higher primates is largely dependent on negative and positive feedback mechanisms by both steroidal and non-steroidal substances of the ovaries which regulate the function of hypothalamo-pituitary system. To gain insights into the role of INH A, the non steroidal ovarian hormone in the feedback control of pituitary FSH secretion, studies were conducted to examine the interrelationships of hormones throughout the menstrual cycle of the bonnet macaque. The findings of chapter II provide a detailed description of endocrine hormone profile during the menstrual cycle of the bonnet macaques with special attention to the feedback role of INH A on pituitary FSH secretion. To characterize the endocrine profile of different hormones, both ovarian (E2, P4, INH A) and pituitary (FSH, LH) hormones were measured daily for more than 40 days. To further examine the site of secretion of INH A and its relationship with pituitary FSH dynamics, surgical lutectomy and pharmacological induction of luteolysis employing the third generation GnRH R antagonist, Cetrorelix (CET) studies were carried out in the subsequent experiments. The results obtained from these studies suggest that INH A and P4 secreted from the CL during luteal phase regulate pituitary FSH secretion. The selective rise in FSH observed during the late menstrual cycle and during menstruation (referred to as luteo-follicular transition), as has been reported previously in higher primates, considered necessary for initiation of follicular growth and recruitment of follicles for ensuing menstrual cycle was characterized in the monkey. Surgical lutectomy and induction of luteolysis by CET experiments suggested that increased GnRH secretion is essential for this selective rise in FSH following withdrawal of inhibition by P4 and INH A. In clinical cases of reproductive ageing, the shortened follicular phase in human females has been identified to be the result of occurrence of early onset of FSH rise during the luteal-follicular transition period. The cause(s) of declining fertility with age in women who still have regular menstrual cycles are not clear, but issues of relationship between dysregulation of selective FSH rise in the late luteal phase and associated infertility could be examined using bonnet monkey as a model system. INH A is secreted in significant quantities by CL in higher primates and the feto placental unit suggesting its importance during fertility and pregnancy. Apart from the negative feedback regulation of pituitary FSH secretion, the complete repertoire of actions of this hormone during pregnancy is yet to be fully understood. The data presented in this thesis is the first comprehensive report showing the endocrine hormone profile of gonadotropins and ovarian hormones including INH A throughout the menstrual cycle of the bonnet macaque. The characterization of INH A profile in bonnet monkey will be of significant value for studies examining the role of INH A in higher primates. Dimeric inhibin has been suggested to be important for regulation of fertility and reproductive functions. Also, inhibin-α (one of the subunits of the dimeric protein) knock out mice model has provided convincing evidence that it acts as a tumour suppressor. A great deal of new information has been generated in recent years regarding the potential clinical usefulness of monitoring inhibin levels in blood and biological fluids in gynaecological diseases, pathological pregnancies and other disorders. Emerging clinical roles of inhibin have made INH A an important candidate molecule to study its molecular regulation. The results presented in chapter II suggested that LH regulates luteal INH A secretion (induction of luteolysis by CET administration experiment). As a first step towards understanding molecular regulation of inhibin-α expression in the macaque CL, in silico promoter analysis of macaque inhibin-α was performed and it revealed several transcriptional factor binding sites that were conserved across species. In rats FSH up regulates while preovulatory LH surge suppresses inhibin-α mRNA expression in the ovary and this suppression has been suggested to be necessary for occurrence of secondary FSH surge during metestrus. To address differential regulation of inhibin-α by LH and FSH in rat ovary during the periovulatory period, studies employing immature rats were carried out and the results are presented in chapter III. The results suggest that immature rat ovaries respond to exogenous gonadotropins in terms of LH signaling (cAMP production), luteinization (P4 production) and as well induction of ICER expression required for repression of inhibin-α subunit expression. PDE4 inhibitor (rolipram) treatment enhanced the ovarian cAMP concentrations suggesting that PDE4 play a major role in controlling intraovarian cAMP concentrations in rat ovaries. However increased cAMP concentrations did not appear to up regulate the ICER expression at the time point examined in this study. In higher primates time course of second FSH surge and continued synthesis and secretion of INH A in the CL are different from non primate species. In the monkey, the second FSH rise occurs during the late luteal phase and experiments have been carried out to examine the regulation of inhibin-α subunit expression by ICER. Expressions of ICER (mRNA/protein) and INH A were examined during different stages of CL and the results indicated no clear inverse relationship between the ICER and inhibin-α mRNAs. With no conclusive role for the ICER in regulating luteal inhibin-α observed in the study, the role of transcriptional activators in the regulation of inhibin-α like GATA4, SF-1, β-catenin were further examined. Since luteal INH A secretion was dependent on pituitary LH as determined earlier in chapter II, expressions of transcriptional activators were examined in CL of different stages and also during induced luteolysis and the results are described in chapter IV. In conclusion, our results indicate cross talk between WNT, cAMP and P38 MAP kinase signaling pathways in the regulation of luteal INH A secretion. The pituitary gonadotropin, LH, is the primary luteotropin in primate species acting to maintain the structure and function of the CL during the menstrual cycle. However whether the actions of LH are direct or mediated by local factors such as P4 remain unknown. Moreover, P4 secretion which is dominant during luteal phase has any role in regulating CL structure and function is not clearly defined. To address these and issues concerning P4 actions, initially, experiments were performed in the rat model to study the importance of P4 in the regulation of ovarian functions. An antiprogestin, RU486, was employed as a tool to uncover the PR regulated pathways during ovulation in rats and the findings are presented in the chapter V. The results indicated that blockade of PR action by RU486 during gonadotropin-induced superovulation resulted in inhibition of follicular rupture and ovulation in immature rats. Further to understand the downstream effectors of PR action, and to identify the candidate target genes of PR activation, semi-quantitative RT-PCR and western blot analyses were performed. The results obtained indicated that betacellulin, a member of EGF family and MMP-9 a proteolytic enzyme, were markedly repressed in response to RU486 treatment in rat ovaries. Also, the down stream pathway of EGF signaling leading to activation of ERK was markedly repressed in RU486 treated ovaries. It was next examined what role the P4/PR system has in the regulation of CL structure and function. Surprisingly, PR expression is absent in CL of rats, while it is present in higher primates. Experiments were carried out to examine intracrine actions of P4 in the regulation of CL structure and function in monkeys. The recently reported model system of induced luteolysis yet capable of responsive to trophic support from the laboratory provided an ideal opportunity to examine direct effects of P4 on structure and function of CL in the monkey. A series of pilot experiments were carried out in monkeys experiencing summer amenorrhea, to determine dose and mode of administration of exogeneous P4 to simulate mid luteal phase circulating P4 concentrations in monkeys subjected to induced luteolysis. Based on the results of pilot experiments, implantation of Alzet pumps containing 97.5mg of P4 was selected for maintaining mid luteal phase P4 concentrations. The microarray data of induced luteolysis previously deposited by the laboratory in NGBI’s gene expression omnibus were mined for identification and validation of differentially expressed genes of PR and its target genes following LH depletion and LH replacement experiments. Expressions of PR, PR cofactors and expressions of PR downstream target genes through out the luteal phase and in CL from day1 of menses were also examined. Analysis of expressions of genes revealed that of the 45 genes identified to be regulated by LH treatment, 4 genes were found to be responsive to P4, and 14 were identified to be responsive to both P4 and LH. Morphology of CL tissue sections revealed that P4 treatment appeared to have reversed the induced-luteolysis changes. In another experiment, implantation of P4 during late luteal phase (i.e., the period of declining P4 concentrations) for 24h caused changes in expressions of genes associated with tissue remodeling and morphology of luteal cells. Taken together, the results suggest that induced luteolysis plus P4 replacement model is suitable for assessing the effects of P4 on CL function. The results also suggest that CL could serve as target tissue for examining the genomic and non genomic actions of P4. In summary, studies carried out in the present thesis provides a comprehensive endocrine hormone profile throughout the menstrual cycle of the bonnet monkey with special emphasis on time course of INH A and FSH secretion which is very useful for future investigations. Studies have been carried out in rats and monkeys with different experimental model systems to address molecular mechanisms underlying inhibin-α regulation in the ovary in general and CL in particular. Experimental findings in monkeys could help elucidate the underlying molecular nature of CL functionality and extrapolate to understand luteal insufficiency and infertility producing conditions in humans. Also different model systems have been validated to examine the actions of P4/PR system in rats and monkeys and more importantly to address the direct effects of P4 upon monkey CL structure and function were established. Future investigations based on findings of these studies should help clarify relative roles for LH and P4 during maintenance of CL function and luteolysis.

Corpus luteum is a transient endocrine structure formed from the ruptured ovarian follicle. Its main function is to secrete P4, a pro-gestational hormone, essential for establishment and maintenance of pregnancy in mammals. The modulators of CL structure and function are classified as trophic and lytic factors. The luteotrophic factors include pituitary hormones, growth factors, intra luteal factors and cytokines, while luteolytic factors include PGF2α and oxytocin. The interplay between luteotrophic and luteolytic factors regulates luteal steroidogenesis. The precise timing of expression of various enzymes/proteins required for synthesis and metabolism of P4 constitutes an important process in the overall regulation of CL function. The three hormones LH/CG, E2 and PRL are regarded as luteotrophic factors crucial for control of CL function in mammals. Depending on species, either individually or all three hormones in the form of luteotrophic complex have been shown to participate in the regulation of CL function. In addition to the well-established endocrine role of E2, its secretion is the hallmark of the ovulating follicle, has an important role in the intraovarian growth, differentiation and survival of cells. Chapter I provides a comprehensive review of literature on CL structure and function with emphasis on factors that influence its growth, development, function and demise in bovines and rodents. In Chapter II, studies have been carried out to examine 20α-HSD expression and its activity in the CL of buffalo cow. During induced and spontaneous luteolysis, rapid fall in circulating P4 is one of the early signs of initiation of luteolytic process in several species. In rodents, it is well recognized that during luteolysis, P4 is catabolized into inactive metabolite, 20α-OHP by the reaction of 20α-HSD enzyme during luteolysis. Experiments were carried out to determine 20α-HSD expression and activity throughout the luteal phase and during induced luteolysis in the buffalo cow. Circulating P4 concentration declined rapidly in response to PGF2α treatment, but HPLC analysis of serum samples did not reveal changes in circulating 20α-OHP levels in buffalo cows. In contrast, pseudo pregnant rats receiving PGF2α treatment showed higher 20α-OHP levels at 24 h post treatment. qPCR expression of 20α-HSD in CL during different stages of luteal phase and PGF2α-treated buffalo cows was carried out and higher expression of 20α-HSD was observed at 3 and 18 h post treatment, but its activity was not altered post PGF2α treatment at other time points examined. The expression of the transcription factor Nurr77 which is involved in increased expression of 20α-HSD increased several fold 3 h post PGF2α treatment similar to the observation in PGF2α-treated pseudo pregnant rats. The results suggested that the synthesis rather than catabolism of P4 appears to be primarily affected by PGF2α treatment in buffalo cows in contrast to increased metabolism of P4 as seen in rodents. In bovines, to date no luteotropic actions for E2 has been demonstrated and whether E2 has direct effect on CL function has also not been reported. Expression of CYP19A1 gene that encodes aromatase enzyme although gets down regulated post ovulation but its expression recovers in the CL and also E2 biosynthesis has been reported in the bovine CL. Recently it was observed that CYP19A1 expression was consistently down regulated following administration of luteolytic dose of PGF2α. Experiments were conducted to examine the expression of ERα and ERβ in the CL throughout the buffalo estrous cycle as well as examined the luteal E2 levels post PGF2α treatment. The results indicated that ER expression was detectable during different stages of CL and that circulating and luteal E2 levels declined post PGF2α treatment. It was hypothesized that decrease in luteal E2 levels leads to down regulation of ER signaling and changes in expression of E2 responsive genes in the CL. To test the hypothesis, 89 genes which were regarded as E2 responsive genes were selected and the previously published global gene expression data of the buffalo CL was mined for E2 responsive genes. It was observed that 57 of 89 genes regarded as E2 responsive genes were found to be differentially expressed. Since non pregnant buffalo CL is not regarded as major site of E2 production, to validate the authenticity of differentially E2 expressed genes post PGF2α, CL of another species, the macaque, which is known to secrete abundant E2 was included for the analysis. Incidentally, the global gene expression data for the PGF2 α treated macaques (in which CYP19A1 gene expression also gets down regulated) has previously been reported from the laboratory. Here again, it was observed that nearly 79 of 89 genes were identified to be differentially expressed. To further determine the consequences of decreased ER signaling, molecules associated with survival and apoptosis were examined. The results indicated decreased expression (both mRNA and protein levels) of Akt, Bax and Bcl-2 genes. The results suggested an important role for E2 on CL function in the buffalo cow. In Chapter III, several experiments were conducted in another model system, pregnant rat, in which aromatase expression and therefore E2 production is high in the CL. Experiments were conducted to examine the effects of E2 inhibition and E2 replacement on the expression of genes. For this purpose, pregnant rats were treated with a specific aromatase inhibitor on day 12-15 of pregnancy. Together with AI, exogenous E2 was administered to another group of pregnant rats. The CL collected from different groups of rats on day 16 of pregnancy was subjected to microarray analysis. The analysis post validation of microarray data has shown that clusters of genes could be segregated into various pathways involving luteal steroidogenesis, immune system, various growth factors and apoptotic processes, all directed towards the regulation of CL function. The involvement of E2 in luteal cell proliferation and lipid deposition well corroborated with protein levels for cyclin D1 and ki67 and the results of oil red O staining, respectively. There have been reports implicating PI3K/Akt signaling in cyclin D1 accumulation, but mechanism of action does not appear to involve transcriptional activation of cyclin D1. The results of the present study indicate a decrease in cyclin D1 protein levels due to inhibition of PI3K/Akt signaling by AI treatment which is prevented upon administration of E2 during AI treatment. The findings provide a comprehensive overview for the mechanisms associated with the cell survival, progression, etc. The bioinformatics approach provided complete landscape of functional changes affected by the upstream regulators of genes associated with survival and apoptosis. Also, the findings further strengthen the hypothesis of involvement of E2 in the regulation of CL function by way of activation of Akt, the primary mediator of PI3K signaling in the regulation of cellular component that affect cell survival. In the present study, IGFBP5 which was up regulated during luteal inhibition of E2 with AI treatment was selected for further studies. Although IGFBP5 is known to be associated with follicular atresia in the rat ovary, there is limited data for the involvement of IGFBP5 in either a growth stimulatory or inhibitory action on ovarian cells. Based on present findings, a causal link between reduced ERα transcriptional activities resulting in inhibition of Akt/PKB in the presence of IGFBP5 expression could be proposed. Further, the cellular hypertrophy mediated by E2 has been speculated due to increased proliferation of vascular endothelial cells, blood supply and thus nutrients. E2, together with PRL and placental lactogens, regulates steroidogenesis and cell hypertrophy in the rat CL of pregnancy. In CL, the prominent IGFBP5 mRNA expression in different types of luteal cells has not been reported. The mRNA expression for IGFBP5 across the two types of luteal cells showed higher expression in SLC. Hence, in the present study, it has been speculated that prevention of conversion of SLC to LLC due to lack of E2 biosynthesis in presence of AI might be acting as a source for the increased IGFBP5 levels during mid pregnancy in rat CL and brings about changes associated with lack of E2. Various receptor studies on rat CL have demonstrated the lack of progesterone receptor (PR) mRNA expression in the rat CL negating its involvement as an autocrine/paracrine regulator of CL function through an intracellular receptor, but the involvement of non-PR involvement in mediating such mechanism further strengthens the role of ERs. The luteotrophic complex formation in pregnant rat principally by PRL and E2 has been discussed at length in Chapter III. PRL appears to maintain luteal ER content in the CL during rat pregnancy which further determines the luteotrophic and luteolytic actions of E2. Further, study on expression of E2 responsive genes would help in identifying E2 regulating molecules to get a clear picture on the role of E2 in understanding regulation of the CL function. The interaction of E2 with growth factor signaling including the IGF pathway has been well established in different species and this interaction is tightly linked to ERα expression, an observation interpreted as physiological coupling of growth factor and stress signaling pathways. Attempts were made towards understanding cross talk between the E2 signaling and the IGF1 signaling in few experiments carried out in Chapter IV. Based on the results, it can be proposed that a causal link exists between reduced ERα transcriptional activity and inhibition of Akt/PKB in the presence of IGFBP5. The present study has shown the activity of IGF on ERα activity mediated partly via PI3K/Akt pathway. Hence, the finding further speculates that inhibitory effect of IGFBP5 on E2 induced ERα function was due to sequestration of IGF1, possibly present in serum or produced within the cells. Another striking observation was the down regulation of glucocorticoid receptor (GR) gene, NR3C1, in the data of earlier studies [Priyanka, 2009, GEO accession number GSE8371 and Kunal, 2014, GEO accession number GSE27961] and the present study has been compared and discussed in this thesis. Glucocorticoids provide key signals for differentiation of fetal and placental tissues. Therefore, regulation of glucocorticoid access to the placenta and fetus is recognized as an important determinant of prognosis outcome and subsequent development of the postnatal phenotype. Differential regulation of these genes in CL post E2 deprivation and replacement further emphasize the regulation of CL via various biological, cellular and molecular functions. Interestingly, besides transcriptional regulation of IGF axis components, E2 activated ERα also rapidly influence the activity of IGF axis related to signaling proteins in a non-genomic manner, especially by the PI3K/Akt pathway. PI3K/Akt pathway analysis has been carried out in E2 inhibition and replacement experiments. To further confirm the observations of E2 and growth factor interaction, experiments have been set up with exogenous GH for increasing circulating levels of IGF in the system. The findings suggest that the non-genomic signaling pathway activated by the phosphorylation of ERα induced by E2 gets inhibited in the presence of AI result in increased expression of IGFBP5. The reduction in circulating IGF1 in pregnant rats may be associated with the effect on IGFBP, important for determining biological action of IGF1. The changes observed in the present study emphasize the exclusive effects of the IGFBP5 on the CL function brought about perturbations in luteal E2 content. The experiments described in the present thesis aim at understanding the mechanism responsible for decreased serum and luteal P4 post PGF2α treatment in buffalo cows, i.e. whether PGF2α acts on biosynthetic or catabolic process of P4. In the present study, experiments were designed to elucidate the role of E2 in regulation of CL function, since down regulation of CYP19A1 gene mRNA was one of the early events observed in buffalo cows post PGF2α treatment. This line of research work was extended to rodents, a species that secretes high levels of E2 during pregnancy. Genome wide transcriptional changes data revealed differential expression of several E2 responsive genes following E2 inhibition and replacement treatments. The results revealed importance of ER-mediated PI3K/Akt signaling essential for regulation of many transcriptional regulatory molecules in the CL and an interesting involvement of IGFBP5 as a link between E2 and IGF signaling. These findings further provide an insight into the role of IGFBP5 in E2-mediated actions in rat CL during pregnancy. In conclusion, the present findings suggest inhibitory effect of IGFBP5 on E2-induced ERα function and hence, its selection as a target molecule for regulation of CL function and for many beneficial processes involved in anti-carcinogenic properties can be thought of.

Made available in DSpace on 2016-01-26T18:55:35Z (GMT). No. of bitstreams: 1 deyvid.pdf: 251965 bytes, checksum: 4ba035cb4ab8b1227779ef5ba9b57cb3 (MD5) Previous issue date: 2012-09-26 / The white tea in a healthy drink, but this tea can interfere in some growth factors involved in the reproduction and in the metabolism. So, the aim of this work was to verify the effect of the white tea consumption in the metabolism, number of corpora lutea and in the weight of superovulated rats. There were utilized two groups of rats: control (n=30) that received water and the group that received only white tea to drink (n=30). The experiment lasts three months and the liquid and ration consumption were verified. In the end of each month, 10 rats of each group were superovulated with 150 UI/Kg of eCG, and 150UI/Kg of hCG 48 hours later. After 48 hours of the superovulation, the rats were weighted, sacrificed and the ovaries were weighted and the corpora lutea counted, besides the blood of animals were collected for hemogram and serum biochemistry. Não houve diferença no número de corpos lúteos e peso dos ovários entre os grupos. The statistical analysis was ANOVA test followed of Tukey analysis, differences were considered when P<0.05. The group treated with white tea showed higher ingestion of ration (39.63 &#61617; 3.91 g) than control group (37.19 &#61617; 4.00 g; P<0.05). However there wasn t any difference in the animals weight. Moreover, the treated group presented lower cholesterol concentration (32.02 &#61617; 9.62 mg/dL) than control group (76.70 &#61617; 10.41 mg/dL; P<0.05). No one statistical difference was observed in the number of corpora lutea, neither in the ovaries weight between the groups. The conclusion that white tea inhibis the excessive weight gain and decreases the cholesterol concentration; furthermore the white tea does not interfere with number of corpora lutea neither in the ovaries weight of superovulated rats. / O chá branco é uma bebida saudável, porém este chá pode interferir em vários fatores de crescimento envolvidos na reprodução e no metabolismo. Portanto, este trabalho teve como objetivo verificar o efeito do consumo prolongado de chá branco no metabolismo, no número de corpos lúteos e no peso dos ovários de ratas superovuladas. Foram utilizados dois grupos de ratas: controle (n=30) que recebeu água e o grupo que recebeu apenas chá branco para beber (n=30). O experimento durou 3 meses, o consumo de líquido e de ração foi verificado. Ao final de cada mês, 10 ratas de cada grupo eram superovuladas, com 150 UI/Kg de eCG e mais 150UI/Kg de hCG depois de 48 horas. Após 48 horas da superovulação, as ratas foram pesadas, sacrificadas, os ovários foram pesados e os corpos lúteos contados. O sangue dos animais foi colhido para hemograma e bioquímica sérica. A análise estatística foi de ANOVA seguida do teste de Tukey, foram consideradas diferenças estatísticas quando P<0,05. O grupo tratado com chá branco apresentou maior ingestão de ração (39,63 &#61617; 3,91 g) em relação ao controle (37,19 &#61617; 4,00 g; P<0,05), porém não houve diferença no peso dos animais, o grupo tratado apresentou menor concentração de colesterol (32,02 &#61617; 9,62 mg/dL) que o grupo controle (76,70 &#61617; 10,41 mg/dL; P<0,05). Não houve diferença no número de corpos lúteos e peso dos ovários entre os grupos. Conclui-se que o consumo de chá branco evita o ganho excessivo de peso dos animais, diminui a concentração sérica de colesterol e não interfere no número de corpos lúteos e no peso dos ovários de ratas superovuladas.

Made available in DSpace on 2016-07-18T17:53:10Z (GMT). No. of bitstreams: 1 deyvid.pdf: 251965 bytes, checksum: 4ba035cb4ab8b1227779ef5ba9b57cb3 (MD5) Previous issue date: 2012-09-26 / The white tea in a healthy drink, but this tea can interfere in some growth factors involved in the reproduction and in the metabolism. So, the aim of this work was to verify the effect of the white tea consumption in the metabolism, number of corpora lutea and in the weight of superovulated rats. There were utilized two groups of rats: control (n=30) that received water and the group that received only white tea to drink (n=30). The experiment lasts three months and the liquid and ration consumption were verified. In the end of each month, 10 rats of each group were superovulated with 150 UI/Kg of eCG, and 150UI/Kg of hCG 48 hours later. After 48 hours of the superovulation, the rats were weighted, sacrificed and the ovaries were weighted and the corpora lutea counted, besides the blood of animals were collected for hemogram and serum biochemistry. Não houve diferença no número de corpos lúteos e peso dos ovários entre os grupos. The statistical analysis was ANOVA test followed of Tukey analysis, differences were considered when P<0.05. The group treated with white tea showed higher ingestion of ration (39.63 &#61617; 3.91 g) than control group (37.19 &#61617; 4.00 g; P<0.05). However there wasn t any difference in the animals weight. Moreover, the treated group presented lower cholesterol concentration (32.02 &#61617; 9.62 mg/dL) than control group (76.70 &#61617; 10.41 mg/dL; P<0.05). No one statistical difference was observed in the number of corpora lutea, neither in the ovaries weight between the groups. The conclusion that white tea inhibis the excessive weight gain and decreases the cholesterol concentration; furthermore the white tea does not interfere with number of corpora lutea neither in the ovaries weight of superovulated rats. / O chá branco é uma bebida saudável, porém este chá pode interferir em vários fatores de crescimento envolvidos na reprodução e no metabolismo. Portanto, este trabalho teve como objetivo verificar o efeito do consumo prolongado de chá branco no metabolismo, no número de corpos lúteos e no peso dos ovários de ratas superovuladas. Foram utilizados dois grupos de ratas: controle (n=30) que recebeu água e o grupo que recebeu apenas chá branco para beber (n=30). O experimento durou 3 meses, o consumo de líquido e de ração foi verificado. Ao final de cada mês, 10 ratas de cada grupo eram superovuladas, com 150 UI/Kg de eCG e mais 150UI/Kg de hCG depois de 48 horas. Após 48 horas da superovulação, as ratas foram pesadas, sacrificadas, os ovários foram pesados e os corpos lúteos contados. O sangue dos animais foi colhido para hemograma e bioquímica sérica. A análise estatística foi de ANOVA seguida do teste de Tukey, foram consideradas diferenças estatísticas quando P<0,05. O grupo tratado com chá branco apresentou maior ingestão de ração (39,63 &#61617; 3,91 g) em relação ao controle (37,19 &#61617; 4,00 g; P<0,05), porém não houve diferença no peso dos animais, o grupo tratado apresentou menor concentração de colesterol (32,02 &#61617; 9,62 mg/dL) que o grupo controle (76,70 &#61617; 10,41 mg/dL; P<0,05). Não houve diferença no número de corpos lúteos e peso dos ovários entre os grupos. Conclui-se que o consumo de chá branco evita o ganho excessivo de peso dos animais, diminui a concentração sérica de colesterol e não interfere no número de corpos lúteos e no peso dos ovários de ratas superovuladas.

A hiperplasia adrenal congênita (HAC) por deficiência no P450c17 é uma doença de herança autossômica recessiva raramente relatada em pacientes 46, XX. Nosso objetivo foi o de caracterizar o fenótipo e genótipo desta doença rara revendo os dados clínicos, laboratoriais, genéticos, e da função ovariana de pacientes 46,XX de uma coorte brasileira avaliada no HCFMUSP e dos casos já publicados na literatura. Foram avaliados retrospectivamente os dados de 18 pacientes brasileiras pertencentes a 12 famílias avaliadas no HCFMUSP, e revisados os dados de literatura de pacientes de 10 coortes com deficiência da atividade P450c17 (14 pacientes, 6 delas com defeitos no CYP17A1 e 8 com defeitos no POR). Fenótipo: A maioria das pacientes apresentou-se com amenorreia primária (74 %) e 90 % das pacientes não desenvolveram pubarca; 69,5 % das pacientes apresentavam hipertensão arterial no momento do diagnóstico. Observamos uma alta incidência de distúrbios psiquiátricos (76%), como depressão, ansiedade e síndrome do pânico, em nossa coorte, mas não foram encontrados relatos na literatura. O ultrassom mostrou um aumento de pelo menos um dos ovários em 87 % dos pacientes antes do tratamento e macrocistos ovarianos em 65%, 6 pacientes (26%) referiram terem sido submetidas a cirurgia anterior para tratamento de torção ou ruptura de ovário. O tratamento com dexametasona, estrogênio e progesterona resultou em redução efetiva do volume ovariano. Todos os pacientes apresentaram níveis basais elevados de progesterona e LH e redução dos níveis de androgênios. Não observamos correlação entre os níveis de LH, da relação LH/FSH e de progesterona com o volume ovariano nos dois defeitos. Genótipo: O estudo molecular revelou que 17 pacientes de nossa coorte apresentavam mutações inativadoras no gene CYP17A1 e 1 paciente no gene POR. Duas novas mutações foram identificadas no gene CYP17A1, a p.R362H no éxon 6 e a p.G478S no éxon 8. A mutação mais prevalente no CYP17A1 foi a p.W406R identificada em 41 % das nossas famílias. Algumas das mutações no CYP17A1 foram encontradas apenas na coorte brasileira, mas a mutação p.A287P encontrada no gene POR em uma das nossas pacientes é a mais prevalente na literatura. Em relação à etnia, houve um predomínio dos defeitos moleculares no CYP17A1 em pacientes chinesas e brasileiras, enquanto que os defeitos no gene POR foram mais relatados nas pacientes europeias e norte - americanas. Em conclusão, a análise retrospectiva do fenótipo, genótipo e morfologia ovariana de trinta e duas pacientes 46,XX portadoras de deficiência da atividade P450c17, nos permitiu destacar os seguintes aspectos: a importância da dosagem da progesterona basal para este diagnóstico, a alta prevalência de aumento dos ovários com a presença de macrocistos ovarianos com risco de torção, e de transtornos psiquiátricos identificados em nossa coorte. Acreditamos que esta revisão possa contribuir para o diagnóstico mais precoce desta rara doença congênita / Congenital adrenal hyperplasia due to P450c17 deficiency is rarely reported on 46,XX patients. Our aim was to characterize the phenotype and genotype of this rare disorder reviewing the clinical, laboratory, genetic and ovarian imaging of 46,XX patients of our cohort and of other already reported cases. We retrospectively reviewed 32 patients with deficiency of P450c17 activity: 18 Brazilian patients belonging to 12 families and 10 cohorts already published (fourteen 46,XX patients, 6 of them with CYP17A1 defects and 8 with POR defects). Phenotype: most patients had primary amenorrhea (74%) and 90% of the patients did not develop pubarche; 69.5% of the patients had blood hypertension at diagnosis. We observed a high incidence of psychiatric disorders such as depression, anxiety and panic (76%) in our cohort but no reports were found in the literature. Ultrasound showed an increase of at least one of the ovaries in 87% of the patients before treatment and ovarian macrocysts in 65% of them; 6 patients (26%) had had previous surgery for twisting or ovarian rupture. Treatment with dexamethasone, estrogen and progesterone resulted in ovarian volume reduction. All patients showed elevated basal progesterone and LH and levels, and decreased androgen levels. There was no correlation between the levels of LH and progesterone and of LH/FSH ratio and the ovarian volume in both defects. Genotype: the molecular study showed that 17 patients from our cohort had inactivating mutation in the CYP17A1 gene and 1 in POR gene. Two novel mutations were identified in the CYP17A1 gene, the p.R362H in exon 6 and p.G478S in exon 8. The most prevalent mutation in the CYP17A1 was the p.W406R, identified in 41% of our families. Some of the CYP17A1 mutations were found only in the Brazilian cohort, but the mutation p.A287P found in the POR gene is the most prevalent in the literature. Regarding the ethnicity of the defects, there was a predominance of Chinese and Brazilian patients with defects in the CYP17A1 whereas defects in POR were most reported in European and North-American subjects.Conclusion: In this data review of thirty-two 46,XX patients with dysfunction of P450c17 activity we characterized the phenotype and genotype of this rare disorder and emphasize: the importance of basal progesterone measurement for this diagnosis, the high prevalence of ovarian macrocysts with risk of twisting, and the psychiatric disorders. We believe that this review may contribute to the early diagnosis of this disorder

A hiperplasia adrenal congênita (HAC) por deficiência no P450c17 é uma doença de herança autossômica recessiva raramente relatada em pacientes 46, XX. Nosso objetivo foi o de caracterizar o fenótipo e genótipo desta doença rara revendo os dados clínicos, laboratoriais, genéticos, e da função ovariana de pacientes 46,XX de uma coorte brasileira avaliada no HCFMUSP e dos casos já publicados na literatura. Foram avaliados retrospectivamente os dados de 18 pacientes brasileiras pertencentes a 12 famílias avaliadas no HCFMUSP, e revisados os dados de literatura de pacientes de 10 coortes com deficiência da atividade P450c17 (14 pacientes, 6 delas com defeitos no CYP17A1 e 8 com defeitos no POR). Fenótipo: A maioria das pacientes apresentou-se com amenorreia primária (74 %) e 90 % das pacientes não desenvolveram pubarca; 69,5 % das pacientes apresentavam hipertensão arterial no momento do diagnóstico. Observamos uma alta incidência de distúrbios psiquiátricos (76%), como depressão, ansiedade e síndrome do pânico, em nossa coorte, mas não foram encontrados relatos na literatura. O ultrassom mostrou um aumento de pelo menos um dos ovários em 87 % dos pacientes antes do tratamento e macrocistos ovarianos em 65%, 6 pacientes (26%) referiram terem sido submetidas a cirurgia anterior para tratamento de torção ou ruptura de ovário. O tratamento com dexametasona, estrogênio e progesterona resultou em redução efetiva do volume ovariano. Todos os pacientes apresentaram níveis basais elevados de progesterona e LH e redução dos níveis de androgênios. Não observamos correlação entre os níveis de LH, da relação LH/FSH e de progesterona com o volume ovariano nos dois defeitos. Genótipo: O estudo molecular revelou que 17 pacientes de nossa coorte apresentavam mutações inativadoras no gene CYP17A1 e 1 paciente no gene POR. Duas novas mutações foram identificadas no gene CYP17A1, a p.R362H no éxon 6 e a p.G478S no éxon 8. A mutação mais prevalente no CYP17A1 foi a p.W406R identificada em 41 % das nossas famílias. Algumas das mutações no CYP17A1 foram encontradas apenas na coorte brasileira, mas a mutação p.A287P encontrada no gene POR em uma das nossas pacientes é a mais prevalente na literatura. Em relação à etnia, houve um predomínio dos defeitos moleculares no CYP17A1 em pacientes chinesas e brasileiras, enquanto que os defeitos no gene POR foram mais relatados nas pacientes europeias e norte - americanas. Em conclusão, a análise retrospectiva do fenótipo, genótipo e morfologia ovariana de trinta e duas pacientes 46,XX portadoras de deficiência da atividade P450c17, nos permitiu destacar os seguintes aspectos: a importância da dosagem da progesterona basal para este diagnóstico, a alta prevalência de aumento dos ovários com a presença de macrocistos ovarianos com risco de torção, e de transtornos psiquiátricos identificados em nossa coorte. Acreditamos que esta revisão possa contribuir para o diagnóstico mais precoce desta rara doença congênita / Congenital adrenal hyperplasia due to P450c17 deficiency is rarely reported on 46,XX patients. Our aim was to characterize the phenotype and genotype of this rare disorder reviewing the clinical, laboratory, genetic and ovarian imaging of 46,XX patients of our cohort and of other already reported cases. We retrospectively reviewed 32 patients with deficiency of P450c17 activity: 18 Brazilian patients belonging to 12 families and 10 cohorts already published (fourteen 46,XX patients, 6 of them with CYP17A1 defects and 8 with POR defects). Phenotype: most patients had primary amenorrhea (74%) and 90% of the patients did not develop pubarche; 69.5% of the patients had blood hypertension at diagnosis. We observed a high incidence of psychiatric disorders such as depression, anxiety and panic (76%) in our cohort but no reports were found in the literature. Ultrasound showed an increase of at least one of the ovaries in 87% of the patients before treatment and ovarian macrocysts in 65% of them; 6 patients (26%) had had previous surgery for twisting or ovarian rupture. Treatment with dexamethasone, estrogen and progesterone resulted in ovarian volume reduction. All patients showed elevated basal progesterone and LH and levels, and decreased androgen levels. There was no correlation between the levels of LH and progesterone and of LH/FSH ratio and the ovarian volume in both defects. Genotype: the molecular study showed that 17 patients from our cohort had inactivating mutation in the CYP17A1 gene and 1 in POR gene. Two novel mutations were identified in the CYP17A1 gene, the p.R362H in exon 6 and p.G478S in exon 8. The most prevalent mutation in the CYP17A1 was the p.W406R, identified in 41% of our families. Some of the CYP17A1 mutations were found only in the Brazilian cohort, but the mutation p.A287P found in the POR gene is the most prevalent in the literature. Regarding the ethnicity of the defects, there was a predominance of Chinese and Brazilian patients with defects in the CYP17A1 whereas defects in POR were most reported in European and North-American subjects.Conclusion: In this data review of thirty-two 46,XX patients with dysfunction of P450c17 activity we characterized the phenotype and genotype of this rare disorder and emphasize: the importance of basal progesterone measurement for this diagnosis, the high prevalence of ovarian macrocysts with risk of twisting, and the psychiatric disorders. We believe that this review may contribute to the early diagnosis of this disorder