Avaliação da reserva ovariana na predição do mau prognóstico reprodutivo: quando e como investigar / Ovarian reserve evaluation in the prediction of poor reproductive prognosis: when and how to investigate?

Muitas vezes, torna-se um grande desafio para o ginecologista a identificação daquelas com maior ou menor chance de concepção. Vários marcadores laboratoriais e ultrassonográficos, conhecidos conjuntamente como testes de avaliação da reserva ovariana, são estudados há décadas com a intenção de se buscar uma ferramenta para a predição do potencial reprodutivo. E, embora ainda se busquem os marcadores ideais para aplicação clínica, mais difícil do que os definir é definir quando eles estão indicados. Este artigo de atualização, assinado pela Comissão Nacional Especializada em Ginecologia Endócrina da Febrasgo, pretende oferecer ao leitor as ferramentas necessárias para o uso racional dos testes de avaliação da reserva ovariana no cotidiano.(AU)

Often, it becomes a great challenge for the gynecologist to identify women with a greater or lesser chance of conception. Several laboratory and ultrasound markers, known jointly as ovarian reserve evaluation tests, have been studied for decades with the intention of seeking a tool for the prediction of reproductive potential. And, while the ideal markers for clinical application are still sought, defining them is as harder as defining when they are indicated. This update article, signed by the National Specialized Committee on Gynecologic Endocrinology, Febrasgo, intends to offer the reader the necessary tools for the rational use of ovarian reserve evaluation tests in daily practice.(AU)

Oocyte distribution within and between ovary lobes is largely homogeneous in Lachnolaimus maximus (Perciformes: Labridae) / La distribución de los ovocitos en y entre los lóbulos ováricos de Lachnolaimus maximus (Perciformes: Labridae) es generalmente homogénea

Abstract:In fish reproduction, previous information of ovary oocyte distribution is necessary, when oocytes quantitative estimates are required to estimate batch or annual fecundity. Heterogeneous oocyte distribution requires a standardized sampling protocol to prevent bias in estimates, whereas homogeneous distribution, allows sampling of any portion of gonads with no risk of bias. We studied gonad oocyte distribution patterns in the hogfish Lachnolaimus maximus population from Southern Gulf of Mexico. For this, 23 mature females were selected from a total of 47 individuals exhibiting visible oocytes in the ovaries. These females were classified by reproductive phase and sub-phase (early developing-ED, developing-D, spawning capable-SC and actively spawning-AS). Six histological sections were taken from the anterior, middle and posterior regions of the left and right ovary lobes of each individual. Digital image processing (AxioVision and Image ProPlus programs) was used to estimate oocyte density per unit area, and for different development stages. Contingency tables were used to analyze oocyte distribution frequencies between the regions of each lobe, and between the lobes of each ovary. This was supported with a Pearson's χ2 test for goodness-of-fit and a replicated G test to confirm distribution heterogeneity. Oocyte stage distribution was homogeneous in almost all 23 females regardless of the ovary lobe. In the left ovary lobe, oocyte distribution was uniform in all three regions sampled regardless the female phase or sub-phase. In the right ovary lobe, oocyte frequencies were similarly uniform for the ED, D and AS phase and sub-phases; nevertheless, during SC phase, some heterogeneity was observed in tertiary vitellogenesis-Vtg3 oocytes, especially in samples from middle and posterior regions of this lobe. Females in AS sub-phase are normally used to estimate batch fecundity in fish such as L. maximus, which has shown to have asynchronous oocyte development and batch spawning. Given the homogeneous oocyte distribution pattern within and between the ovary lobes in females in AS sub-phase, no systematization is required of the gonad histological sampling protocol to estimate species batch fecundity. Nevertheless, due to the heterogeneous Vtg3 oocytes distribution in the right ovary lobe of females in the SC phase, it is best to systematically take sections of any region in the left ovary lobe when conducting a study encompassing all of a species' reproductive aspects. Rev. Biol. Trop. 65 (1): 293-303. Epub 2017 March 01.

ResumenPara el estudio de la reproducción de los peces es necesario, un conocimiento previo sobre la distribución de los ovocitos en el ovario, así como estimaciones cuantitativas del número de ovocitos para estimar la fecundidad por lote o anual. Una distribución heterogénea exige tener un protocolo de muestreo estandarizado para prevenir sesgos en las estimaciones, mientras que una distribución homogénea permite utilizar muestras de cualquier parte de las gónadas sin riesgo de sesgo. Nosotros estudiamos el patrón de distribución de los ovocitos de la población de la doncella de pluma Lachnolaimus maximus del sur del Golfo de México. Para este propósito, 23 hembras maduras fueron seleccionadas de un total de 47 individuos que presentaron ovocitos observables a simple vista en los ovarios. Estas hembras fueron clasificadas según su fase o sub-fase reproductiva (desarrollo temprano-DT, desarrollo-D, aptitud para desovar-AD y desove activo-DA). Seis secciones histológicas fueron realizadas de las regiones anterior, media y posterior de los lóbulos derecho e izquierdo del ovario de cada individuo. Un procesamiento digital de imágenes (AxioVision e Image ProPlus) fue utilizado para estimar la densidad de los ovocitos, en diferentes estadios de desarrollo, por unidad de área. Las frecuencias de distribución de los ovocitos fueron analizadas entre regiones de un mismo lóbulo y entre lóbulos de cada ovario por medio de tablas de contingencia. Este análisis involucró la aplicación de la prueba de bondad de ajuste del χ2 de Pearson y de la prueba de G replicada en el caso de observar una distribución heterogénea. La mayoría de las 23 hembras analizadas presentó una distribución homogénea de los diferentes estadios de ovocito, en cualquier lóbulo considerado. En el lóbulo ovárico izquierdo, la distribución de los ovocitos fue similar en las tres regiones muestreadas, en cualquier fase o sub-fase de las hembras. En el lóbulo ovárico derecho, las frecuencias de los ovocitos fueron semejantes para las hembras en fase y sub-fases de DT, D y DA; sin embargo, durante la sub-fase de AD, una heterogeneidad en el desarrollo de los ovocitos en vitelogénesis terciaria-Vtg3 fue observada, especialmente en las muestras de las regiones media y posterior de este lóbulo. Las hembras en sub-fase de DA son usualmente utilizadas para estimar la fecundidad por lote en las especies de peces como L. maximus, la cual presenta un desarrollo asincrónico de los ovocitos y realiza desoves sucesivos por lote. Debido al patrón de distribución homogéneo de los ovocitos en y entre los lóbulos ováricos de las hembras en sub-fase de DA, no se requiere estandarizar un protocolo de muestreo histológico de las gónadas para estimar la fecundidad por lote de la especie. Sin embargo, debido a la distribución heterogénea de los ovocitos en Vtg3 en el lóbulo ovárico derecho de las hembras en fase de DA, es preferible tomar sistemáticamente secciones de cualquier región del lóbulo ovárico izquierdo cuando se realiza un estudio que incluye todo los aspectos reproductivos de la especie.

Effects of natural environmental conditions on the ovarian histometric dynamics of five striped squirrels (Funambulus pennanti) / Efectos de las condiciones ambientales naturales sobre la dinámica histométrica de ovarios en la ardilla de las palmeras (Funambulus pennanti)

The present study was designed to determine the morphological and histological variations in the female palm squirrel's reproductive organs which inhabit croplands of central Punjab. 16 specimens were collected from 3 different locations of Faisalabad district during different months in order to collect their ovaries samples in order to determine the breeding phenology of this species. Body weight and external body measurements were recorded. Two adult females were autopsied each month and their ovaries were extracted. Tissue were prepared by the tissue paraffin technique and stained by H&E. Seasonal variations in the follicular size and shape were observed. Statistical analysis revealed that values of all parameters of ovaries were significantly (P<0.01) higher during peak folliculogenic phase (January to May) as compared to low folliculogenic phase (November­December). Three reproductive phases were observed according to follicular development i.e., quiescent period (November- December), recrudescence period (January) and peak reproductive phase (February­March). In conclusion of this study, different phases of sexual activities are determined which are controlled by seasonal variations. These variations are in accordance to different physical factors which influences gross anatomical and histological changes of ovaries and folliculogensis of female five-striped northern palm squirrel (Funambulus pennanti) for adjustments in breeding phenology.

El presente estudio fue diseñado para determinar las variaciones morfológicas e histológicas en los órganos reproductores de la ardilla de las palmeras que habita en las tierras de cultivo del centro de Punjab. Dieciséis muestras fueron recolectadas en 3 lugares diferentes del distrito de Faisalabad durante diferentes meses con el fin de obtener los ovarios y determinar la fenología reproductiva de esta especie. Se registraron el peso corporal y las medidas corporales externas. Se realizaron autopsias de dos hembras adultas cada mes y sus ovarios fueron extraídos. Se prepararon los tejidos por técnica de parafina y tinción H & E y se observaron las variaciones estacionales en el tamaño y forma folicular. El análisis estadístico reveló que los valores de todos los parámetros de los ovarios fueron significativamente más altos durante la fase foliculogénica (enero a mayo), en comparación con la fase baja (noviembre-diciembre) (P <0,01). De acuerdo al desarrollo folicular se observaron tres fases reproductivas, el período de reposo (noviembre-diciembre), el período de recrudecimiento (enero) y la fase peak reproductiva (febrero-marzo). En conclusión las diferentes fases de las actividades sexuales definitivamente están controladas por variaciones estacionales. Estas variaciones se ajustan a diferentes factores físicos que influyen en los cambios anatómicos e histológicos de los ovarios y la folículogénesis de la ardilla hembra de las palmeras (Funambulus pennanti) para realizar ajustes en la fenología de su reproducción.

Monitoramento do ciclo estral de cutias (dasyprocta leporina lichtenstein, 1823) através de citologia esfoliativa vaginal e ultrassonografia / Monitoring of the estrous cycle of agoutis (dasyprocta leporina lichtenstein, 1823) by vaginal exfoliative cytology and ultrasonography

O objetivo deste estudo foi monitorar o ciclo estral em cutias (Dasyprocta leporina) criadas em cativeiro no semiárido brasileiro. Durante 70 dias, cinco cutias foram diariamente submetidas a citologia esfoliativa vaginal, e o monitoramento ultrassonográfico ovariano foi realizado a cada três dias. Um total de 8 ciclos estrais foi completamente monitorado, com duração de 28,2±0,7 dias, variando de 24 a 31 dias. Pela citologia esfoliativa vaginal, houve uma predominância de células superficiais nas fases de proestro e estro (P<0,05), seguida da predominância de células intermediárias no metaestro (P<0,05) e de células parabasais no diestro (P<0,05). Por ultrassonografia, não houve diferenças na morfologia ovariana durante as diferentes fases do ciclo estral (P>0,05). Os folículos foram identificados durante as fases estrogênicas (proestro e estro), com diâmetro médio de 1±0,5mm. Em apenas 12,5% das fases luteais, corpos lúteos medindo 1,4±0,9mm foram identificados. Conclui-se que a associação da citologia vaginal e da ultrassonografia ovariana constitui uma alternativa viável para o monitoramento de ciclos estrais e identificação das fases estrogênicas em cutias da espécie Dasyprocta leporina.

The objective of the study was to monitor the estrous cycle in agoutis (Dasyprocta leporina) bred in captivity in Brazilian semiarid. During 70 days, five agoutis were daily subjected to vaginal exfoliative cytology, and the ovarian ultrasound monitoring was conducted every three days. A total of 8 estrous cycles were completely monitored, lasting 28.2±0.7 days, ranging from 24 to 31 days. By vaginal exfoliative cytology, there was predominance of superficial cells at proestrus and estrus phases (P<0.05), followed by the predominance of intermediate cells in the metestrus (P<0.05) and parabasal cells in diestrus (P<0.05). By ultrasound, there were no differences in ovarian morphology during the different phases of the estrous cycle (P>0.05). Follicles during the estrogenic phases (proestrus and estrus) were identified, with an average diameter of 1±0.5mm. In only 12.5% of luteal phases, corpora lutea measuring 1.4±0.9mm were identified. We conclude that the association of vaginal cytology and ovarian ultrasonography is a useful alternative for monitoring the estrous cycle and identifying the estrogenic phases in Dasyprocta leporina.

role of different photoperiods in the activation of the thyroid gland and ovaries of adult and aged rats [Rattus norvegics]

Many animals use the length of the day [photoperiod] to predict and adjust to seasonal changes in the environment through predictive changes in physiology and behaviour. The present work is designed to study the effect of different photoperiod regimes, i.e., short photoperiod 10L:14D, long photoperiod 14L:10D and 12L:12D light-dark cycle [LD] on the structure and hormonal secretion of the thyroid gland and ovaries in adult and aged female rats. The rats were divided into 6 groups, each group contained 6 adults [6 months old] or aged rats [18 months old] and maintained for one month under different photoperiod regimes. Short and long photoperiods increased the activity of the thyroid gland of the adult rats. This gland was full of small follicles lined with simple columnar epithelium. Serum thyroxin levels were extremely higher in long photoperiod group than that observed in 12:12LD and short photoperiod groups. In contrast short photoperiod induced an increase in serum thyroxin level in the aged rats. Short photoperiod and 12:12 LD cycle enhanced gonadal activity of the adult rats. The ovary contained clusters of primordial follicles, different stages of ovarian follicles, and, few numbers of atretic follicles. In the groups of the aged rats all the stages of the ovarian follicles exhibited signs of atresia in such groups. LH, FSH, progesterone and estrogen were increased mainly in the control group in adult and aged rats compared to the photoperiod groups. From the present result we could conclude that: 12:12 light dark cycle enhanced the structural and functional activities of ovaries in adult rats, while aging decreased their activities. In contrast, long photoperiod induced thyroid gland activities in adult rats while short photoperiods activated the gland of aged rats

[ review on the activity of hormones and growth factors after ovarian tissue transplantation]

The majority of cancer treatments are invasive. Gonadal injuries cause reductions in fertility which results in lack of hope for conception in cancer patients and frustration for their partners. Fortunately, current advancements in cryopreservation and transplantation sciences regarding fertility preservation lead to cryostorage of gonads and preservation prior to the onset of chemo- and radiotherapy treatments. Accordingly in women, the main goal of ovarian cryopreservation is establishment of fertility and hormonal cycle restoration after auto-transplantation. Although the history of ovarian transplantation dates back to the 19[th] century, there are reports of live human births following ovarian tissue cryopreservation and transplantation since the past 100 years. Despite this success and additional research in the field of ovarian cryopreservation and transplantation, numerous questions remain unanswered. Among these questions, growth factors and hormonal changes because of their effects on follicular function appear to be more important during ovarian tissue transplantation. This review attempts to address hormones and growth factor functions with the specifics of ovarian cryopreservation and auto-transplantation

Locus coeruleus lesions and PCOS: role of the central and peripheral sympathetic nervous system in the ovarian function of rat

"Polycystic ovary syndrome [PCOS] is a complex endocrine and metabolic disorder associated with ovulatory dysfunction". "Autonomic and central nervous systems play important roles in the regulation of ovarian physiology". The noradrenergic nucleus locus coeruleus [LC] plays a central role in the regulation of the sympathetic nervous system and synaptically connected to the preganglionic cell bodies of the ovarian sympathetic pathway and its activation is essential to trigger spontaneous or induced LH surges. This study evaluates sympathetic outflow in central and peripheral pathways in PCO rats. Our objectives in this study were [1] to estimate LC activity in rats with estradiol valerate [EV]-induced PCO; [2] to antagonized alpha2a adrenoceptor in systemic conditions with yohimbine. Forty two rats were divided into two groups: 1] LC and yohimbine and 2] control. Every group subdivided in two groups: eighteen rats were treated with estradiol valerate for induction of follicular cysts and the remainders were sesame oil groups. Estradiol concentration was significantly augmented by the LC lesion in PCO rats [p<0.001], while LC lesion could not alter serum concentrations of LH and FSH, like yohimbine. The morphological observations of ovaries of LC lesion rats showed follicles with hyperthecosis, but yohimbine reduced the number of cysts, increased corpus lutea and developed follicles. Rats with EV-induced PCO increased sympathetic activity. LC lesion and yohimbine decreased the number of cysts and yohimbine increased corpus lutea and developed follicles in PCO rats

Follicular-stromal interaction in mare ovary during the reproductive cycle / Interacción estroma-folículo en el ovario de la yegua durante el ciclo reproductivo

Ovogenesis and foliculogenesis in the mare differ from other farm animals and therefore, gamete manipulation, in vitro fertilization and embryo transfer have been very difficult. The histologic traits of ovaries from 12 mares, in estrus and diestrus were studied. Number and size of follicules and corpora lutea were recorded. The ovarian stroma was evaluated using the pricosirius technique for collagen. A simple morphometric analysis was done using computerized scanner programs. During late estrus, one dominant follicle (46 +/- 4mm) is seen. The surrounding stroma contains collagen I and III. By the end of the estrus, one hemorrhagic follicle is seen, plus one or two small follicle (2 mm in size). In early diestrus there is a corpus luteum (43-60 mm) and some antral follicles (6 +/- 1). Collagen I forms strands inside the corpus luteum and predominates in the perifollicular ovarian stroma. Modifications of the extracellular matrix may change cell function by way of integrines, the matrix being in turn modified by hormones and other tissular factors. Estrogens can be related to collagen III predominance in ovarian stroma whereas progesterone is associated with increased collagen I. Therefore, follicular-stromal interactions are important in ovarian histophysiology.

La ovogénesis y foliculogénesis en la yegua, difiere de otros animales de granja, y por lo tanto, la manipulación de gametos, fertilización in vitro y transferencia de embriones ha sido muy dificultoso. Se estudiaron las características histológicas de ovarios de 12 yeguas durante el estro y el diestro. Se registró el número y tamaño de los folículos y cuerpos lúteos. Se evaluó el estroma ovárico usando la técnica picrosirius para colágeno. Se realizó un análisis morfométrico simple utilizando un programa escáner computarizado. Durante el estro tardío, se observó un folículo dominante (46 +/- 4 mm). El estroma circundante contiene colágeno I y III. Para el final de este período, se aprecia un folículo hemorrágico, más uno o dos pequeños folículos (2 mm de tamaño). En diestro temprano, hay un cuerpo lúteo (43 - 60 mm) y algunos folículos antrales (6 +/- 1). El colágeno I forma filamentos dentro del cuerpo lúteo y predomina en el estroma ovárico perifolicular. Modificaciones en la matriz extracelular pueden cambiar la función celular vía integrinas, la matriz está a su vez modificada por las hormonas y otros factores tisulares. Los estrógenos se pueden asociar a la predominancia de colágeno III en el estroma ovárico, mientras que la progesterona se asocia con un aumento de colágeno I. Por lo tanto, las interacciones estroma-folículo son importantes en la histofisiología del ovario.

Fisiología reproductiva y cambios evolutivos con la edad de la mujer / Reproductive physiology and evolutive changes with women age

Los cambios fisiológicos con la edad, son consecuencia de cambios sistémicos generales producidos por el envejecimiento, entre los que se destacan cambios a nivel de los mecanismos de defensa relacionados con las especies reactivas del oxígeno (ROS) y alteraciones de la microcirculación. Los cambios evolutivos ováricos con la edad, se deben esencialmente a una depleción de la población folicular, la cual al disminuir por debajo de 1000 folículos, hace perder la capacidad del ovario de ciclar en forma normal, con la consiguiente disminución en los niveles de esteroides circulantes y sus efectos secundarios en los órganos efectores. Asociado a la disminución de la población folicular, se producen cambios en la calidad ovocitaria, los cuales determinan la disminución progresiva de la fertilidad en mujeres mayores de 35 años. Entre los cambios más frecuentes se observan aumento de aneuploidias, disfunciones mitocondriales, cambios de la microcirculación y disminución de la capacidad defensiva sobrelas ROS, entre otros.

The physiological changes with age are the consequence of systemic general changes produced by aging, where changes of the defense mechanisms related to ROS and microcirculation alterations are highlighted. The evolutive changes produced by age in the ovary are related to the follicular depletion. When the ovarian follicle population decrease below 1000 follicles, the ovary loses its capacityto cycle normally. As a consequence, the levels of circulating steroids diminished, producing negatives effects on secondary steroidal organs. In association with the follicular depletion, there are changes in oocyte quality, which determine the progressive diminution of fertility in women older than 35 years. The most frequent changes observed are an increase of the aneuploidies, mitochondrial dysfunction, microcirculation changes and a diminution of the defense capacity to ROS among others.

Morphological features and vascularization study of caprine cyclic corpus luteum / Características morfológicas e estudo da vascularização do corpo lúteo cíclico de cabras ao longo do ciclo estral

Corpus luteum is a temporary endocrine gland that regulates either the estrous cycle and pregnancy. It presents extreme dependency on the adequate blood supply. This work aims to evaluate goat corpus luteum (CL) vascular density (VD) over the estrous cycle. For that purpose, 20 females were submitted to estrus synchronization/ovulation treatment using a medroxyprogesterone intra-vaginal sponge as well as intramuscular (IM) application of cloprostenol and equine chorionic gonadotrophine (eCG). After sponge removal, estrus was identified at about 72hs. Once treatment was over, female goats were then subdivided into 4 groups (n=5 each) and slaughtered on days 2, 12, 16 and 22 after ovulation (p.o). Ovaries were collected, withdrawn and weighted. CL and ovaries had size and area recorded. Blood samples were collected and the plasma progesterone (P4) was measured through RIA commercial kits. The VD was 24.42±6.66, 36.26±5.61, 8.59±2.2 and 3.97±1.12 vessels/mm² for days 2, 12, 16 and 22 p.o, respectively. Progesterone plasma concentrations were 0.49±0.08, 2.63±0.66, 0.61±0.14 and 0.22±0.04ng/ml for days 2, 12, 16 e 22 p.o, respectively. Studied parameters were affected by the estrous cycle phase. Values greater than 12 p.o were observed. In the present work we observed that ovulation occurred predominantly in the right ovary (70 percent of the animals), which in turn presented bigger measures than the contra lateral one. There is a meaningful relationship between the weight and size of the ovary and these of CL (r=0.87, r=0.70, respectively, p<0.05). It is possible to conclude that morphology of goat's ovaries and plasma progesterone concentration changed according to estrous cycle stages. We propose these parameters can be used as indicators of CL functional activity.

O corpo lúteo é uma glândula endócrina temporária que regula tanto o ciclo estral quanto a prenhez, apresentando extrema dependência de aporte sanguíneo adequado. Objetivaram-se avaliar mudanças morfométricas dos ovários e densidade vascular (DV) dos corpos lúteos (CL) de cabras ao longo do ciclo estral (AOLC). Vinte animais foram submetidos ao tratamento para indução/sincronização do estro, usando esponjas intravaginais commedroxiprogesterona, associadas a aplicações intramusculares de cloprostenol e gonadotrofina coriônica eqüina. Após remoção das esponjas, o estro foi identificado em aproximadamente de 72h. Concluído o tratamento, as cabras foram subdivididas em 4 grupos (n=5 cada) para abate nos dias 2, 12, 16 e 22 após ovulação (p.o.). Posteriormente, foram retirados os ovários e realizadas as mensurações de peso, tamanho e área do órgão e dos CL. Amostras de sangue foram coletadas e a progesterona sérica (P4) mensurada utilizando-se RIA convencional. A DV média dos CL AOLC foi 24,42±6,66; 36,26±5,61; 8,59±2,2 e 3,97±1,12 vasos/mm2 para os dias 2, 12, 16 e 22 p.o., respectivamente. A concentração média de P4 foi de 0,49±0,08; 2,63±0,66; 0,61±0,14 e 0,22±0,04ng/ml para os dias 2, 12, 16 e 22 p.o., respectivamente. Os parâmetros em estudo também se mostraram afetados pela fase do ciclo estral, sendo observados os maiores (p < 0,05) valores no dia 12 p.o. Neste experimento, a ovulação ocorreu predominantemente no ovário direito (70 por cento dos animais), o qual apresentou medidas maiores que o contralateral. Observou-se ainda alta correlação significativa entre o peso do ovário e o do CL (r=0,87; p<0,05) e entre o tamanho destes órgãos (r=0,70; p<0,05). Conclui-se que, a morfologia dos ovários de cabras e a concentração sérica de progesterona variam em função da fase do ciclo estral e podem ser utilizadas como parâmetro na avaliação funcional do órgão.

The Use of Tenaculum During Intrauterine Insemination May Not Affect the Pregnancy Outcome

PURPOSE: The association between tenaculum application to the cervix just before embryo transfer and lower pregnancy rate has been reported. However, studies on the use of tenaculum in intrauterine insemination (IUI) cycles are not available. This study attempted to investigate whether the use of tenaculum affects the clinical outcomes of IUI. MATERIALS AND METHODS: Two hundred and thirty three CC/hMG/IUI cycles of one hundred and forty three couples were recruited at Seoul National University Hospital from October 2006 to December 2008. Mock insemination and IUI with or without tenaculum application to the cervix were also performed, and clinical pregnancy rate was compared. RESULTS: The incidence of difficult mock insemination at the beginning of cycle was higher in the tenaculum use group (p < 0.001). No significant statistical difference in the clinical pregnancy rate was observed between the groups with or without tenaculum application (12.1% vs. 18.5%; p = 0.175), which was not influenced by the difficulty of mock insemination. CONCLUSION: The use of tenaculum during IUI may not affect the pregnancy outcome. Our results need to be confirmed by a prospective study in a larger population.

Leptina y función ovárica / Leptin and ovarian function

La leptina, producto del gen de la obesidad (ob), es una proteína sintetizada principalmente por el tejido adiposo y secretada al torrente sanguíneo. Está caracterizada como una hormona que regula el apetito y el gasto energético a través de su acción a nivel central. Sin embargo, esta hormona tiene muchos otros efectos como consecuencia de su interacción directa en sistemas periféricos. Esta proteína media sus efectos a través de receptores específicos, que presentan distintas isoformas y están ampliamente distribuidos en diversos tejidos, incluyendo el eje hipotálamo-hipófisis-ovario. La leptina ejerce diferentes efectos sobre la reproducción en múltiples sistemas y el principal sitio de acción depende de los niveles circulantes de la proteína y de la expresión de sus receptores. Se necesita un estrecho y específico rango de concentración de leptina para mantener una función reproductiva normal, ya que concentraciones por debajo o superiores a esos umbrales interfieren en la capacidad reproductiva tanto a nivel central como ovárico. Es capaz de ejercer efectos directos sobre la función ovárica, tanto inhibitorios como estimulantes. Varios estudios demostraron que la leptina es capaz de regular la expresión de sus propios receptores a lo largo del eje hipotálamo-adenohipófisis-ovario en forma diferencial según el tejido y el estado hormonal. Por lo tanto, bajos o altos niveles de leptina, producidos por diferentes patologías o desequilibrios metabólicos como la obesidad, la diabetes o la anorexia nerviosa, pueden alterar la función ovárica a través de una regulación positiva o negativa sobre sus receptores.

Avaliação da reserva ovariana: métodos atuais / Assessment of ovarian reserve: recent methods

A avaliação do potencial reprodutivo de uma mulher traz grandes dificuldades na busca de um parâmetro fidedigno. Entre os testes disponíveis, incluem-se a biópsia ovariana, a ultrassonografia transvaginal, exames hormonais e testes com hormônios exógenos. Entretanto, nenhum deles demonstra resultados satisfatórios na investigação de pacientes inférteis. Mais recentemente, o Hormônio Antimülleriano tem sido considerado um marcador promissor dessa reserva. Esta revisão teve o objetivo de analisar cada um dos testes disponíveis, evidenciando vantagens e riscos potenciais, assim como sua real aplicabilidade na rotina ginecológica. Para isso, realizou-se uma revisão da literatura de 1998 até 2007, através do PubMed. Como resultado, observou-se que a dosagem de FSH, a contagem de folículos antrais e, principalmente, a dosagem de Hormônio Antimülleriano constituem os parâmetros mais promissores para esse objetivo. Novos estudos, entretanto, se fazem necessários para a melhor determinação desses resultados.

The assessment of a woman's reproductive potential brings difficulties in the search for a trusting parameter of this function. The tests available are ovarian biopsy, transvaginal pelvic ultrasound, hormonal parameters and tests using exogenous hormones. However, none of those shows satisfactory results for the investigation of infertile patients. Recently, Anti-Müllerian Hormone emerged as a promising marker of ovarian reserve. The objetive of this literature review is to analyse each one of the tests available, focusing in their potential advantages and risks, as well as its real applicability in gynecological practice. In order to accomplish that, articles from 1998 to 2007 were searched in PubMed. The results show that serum FSH, antral follicle count and mainly serum Anti-Müllerian Hormone are promising parameters for this aim. Other studies, however, are needed so this conclusion can be clarified.

Avaliação da reserva ovariana / Assessment of ovarian reserve

A diminuição da reserva ovariana é um processo de declínio da fecundidade associado tanto à depleção folicular dos ovários quanto à piora da qualidade oocitária que ocorre com o avanço da idade feminina. Na tentativa de avaliar a reserva ovariana, são realizados testes, como a medida do volume ovariano, a contagem dos folículos antrais do ovário, a dosagem basal do hormônio folículo-estimulante, a dosagem de inibina-B, do hormônio anti-mülleriano e o teste do clomifeno. A contagem folicular antral associada ao hormônio folículo-estimulante basal parece ser a abordagem de menor custo na avaliação do casal infértil, mas a contagem folicular antral associada aos níveis séricos de anti-mülleriano tem o maior poder preditivo de sucesso no tratamento de reprodução assistida.

The ovarian reserve diminution is a fecundity decrease process associated both to the follicular depletion of the ovaries as well as to the decline of the oocyte quality that happens due to female ageing. Tests like the ovarian volume measurement, the ovarian basal antral follicle counting, the basal follicle-stimulating hormone level, the dosage of the inhibin B and of the anti-Müllerian hormone, and the clomiphene citrate challenge test are made trying to evaluate the ovarian reserve. The basal antral follicle count associated to the basal follicle-stimulating hormone dosage seems to be the lowest cost approach in the evaluation of the infertile couple, but the basal antral follicle count associated to the serum anti-Müllerian levels has higher success predictive power in the assisted reproduction treatment.

Avaliação de protocolos para indução de inatividade ovariana em gatas domésticas / Evaluation of protocols for induction of inactivity ovarian in domestic cats

Os índices de prenhez após inseminação artificial em felídeos selvagens não são satisfatórios devido ao variável ambiente endócrino após a estimulação com gonadotropinas. O objetivo deste estudo consistiuem aumentar a taxa de sucesso em programas de inseminação artificial em gatas domésticas (animal modelo). As fêmeas (n=9) foram divididas em três grupos, cada um com três animais, sendo: 1) controle(C), somente 200 UI eCG/ 100 UI hCG ; 2) levonorgestrel oral (L)(0,075 mg) durante 37 dias + eCG/hCG; 3) etonogestrel (E), implante subdérmico durante 37 dias + eCG/hCG. Foram submetidas ao exame laparoscópico 29-39 horas após a administração de hCG para verificação da resposta ovariana e realização de esfregaço vaginal para monitoração da fase do ciclo estral. Foram coletadas amostras de fezes 60 dias antes e 60 dias após o tratamento com gonadotropinas para dosagem hormonal de estrógenos. Os resultados foram avaliados através do Teste ANOVA. Os níveis de significância mostraram que o Grupo E, em contraste com o Grupo C e o Grupo L, apresentou inibição satisfatória das concentrações de estrógenos durante a sua utilização. O grupo L não apresentou inibição ovariana durante o tratamento e diferença significativa em relação ao Grupo C. No exame laparoscópico todas as fêmeas dos grupos C, L e E apresentaram folículos e 77% das fêmeas apresentaram corpo lúteo. Também apresentaram células epiteliais superficiais anucleadas e nucleadas características de estro. Concluiu-se que a utilização de implantes de etonogestrel em gatas domésticas mostrou-se eficaz, possibilitando asua utilização prévia aos programas de inseminação artificial, aspiração folicular e também para a contracepção.

Reproductive success in endangered captive small felids species is veryl ow. Due to great variability in endocrine environment post gonadotropin treatment, after artificial insemination pregnancy rates are very low. Nowadays, ovarian activity controll improves the AI success in many species. In this study, new protocols were compared to improve the fertilization rates in artificial insemination programs in domestic cat. Female domestic cats were divided in three treatments: 1) control (eCG/hCG); 2) levonorgestrel (0.075 mg) orally during 37days + eCG/hCG; 3) etonogestrel subdermal implant during 37days + eCG/hCG: Laparoscopies were done 29-39 hours post hCG treatment to verify ovarian activity. Vaginal swabs were collected at laparoscopic procedures. Fecal samples were colected 60 days before, during and 60 days after the gonadotropin treatment for estradiol assay. Means comparisons were done by ANOVA test. Results demonstrated that etonogestrel (implant) and not oral levonorgestrel successfully suppressed ovarian activity. The levonorgestrel group didnot show ovarian inactivity during the administration, presenting oestradiol peaks and without significative diference comparing to control group. All females presented anuclear and nuclear superficial vaginal epithelial cells at laparoscopies. In conclusion, the etonogestrel implant used in the domestic cat was efficient and can be used previous to gonadotropin protocol in artificial insemination programs, follicular aspiration and contraception

Oocyte structure and ultrastructure in the Mexican silverside fish Chirostoma humboldtianum (Atheriniformes: Atherinopsidae)

the structural and ultrastructural features of gonads from endemic Mexican fish have received scarce attention. This study describes the histological and ultrastructural characteristics of the oocyte in Chirostoma humboldtianum. The ovary is asynchronic, and as such, most phases of oocyte development are found in the same ovary. The complete process of oogenesis was divided in five stages: oogonium and folliculogenesis, primary growth, cortical alveoli and lipid inclusions, vitellogenesis and maturation. The presence of big filaments, which appear at the end of primary growth, induces some common follicular adaptation. During primary growth, abundant ribosomes, rough endoplasmic reticulum, and mitochondria are grouped in the cytoplasm. At the end of this stage, the Z1 layer of the chorion is developed, while microvilli start to be evident as well. In the cortical alveoli and lipid droplets phase, intense PAS positive vesicles, some of them containing nucleoid material, are observed in the peripheral cytoplasm and the lipid droplets take a more central position. In vitellogenesis, the proteic yolk accumulates in a centripetal way while the chorion is completely formed. In maturation, the germinal vesicle migrates to the animal pole, meiosis is restored, and there is nuclear breakdown. The oocyte increases its size and holds some oil droplets and a big fluid mass of yolk. On the outside, filaments surround the oocyte completely. Rev. Biol. Trop. 56 (4): 1825-1835. Epub 2008 December 12.

Los aspectos estructurales y ultraestructrurales de las gónadas de peces mexicanos endémicos han sido poco estudiados. En el presente trabajo reportamos las características histológicas y ultraestructurales del ovocito de Chirostoma hulmboldtianum. El ovario es de tipo asincrónico, por ende, la mayoría de las fases del desarrollo del ovocito pueden ser encontradas en el mismo ovario. El desarrollo del ovocito fue dividido en cinco etapas: ovo-gonia y foliculogénesis, crecimiento primario del ovocito, inclusiones lipídicas y gránulos corticales, vitelogénesis y maduración. La presencia de grandes filamentos que aparecen al final de la etapa de crecimiento primario, inducen adaptaciones foliculares. Durante el crecimiento primario, en el citoplasma se encuentran abundantes ribosomas, retículo endoplásmico rugoso y agrupamientos de mitocondrias. Al final de esta etapa, inicia el desarrollo de la capa Z1 del corion, comenzando a ser evidentes las microvellosidades del ovocito. Durante la etapa de inclusiones lipídicas y gránulos corticales, vesículas PAS positivas, algunas de ellas con material nucleoide, se ubican en la periferia del ovocito, mientras que las que contienen material graso toman una posición más central en la célula. Durante la vitelogénesis se presenta una acumulación de vitelo protéico en un sentido centrípeto; durante esta etapa, el corion está completamente formado. En la maduración, la vesicular germinal migra hacia el polo animal, se reinicia la meiosis y se rompe la envoltura nuclear. El ovocito incrementa su tamaño y en el citoplasma se pueden observar algunas gotas de grasa y el vitelo se presenta como una gran masa acuosa. En el exterior, los filamentos rodean completamente al ovocito.

Oocyte structure and ultrastructure in the Mexican silverside fish Chirostoma humboldtianum (Atheriniforme: Atherinopsidae)

The structural and ultrastructural features of gonads from endemic Mexican fish have received scarce attention. This study describes the histological and ultrastructural characteristics of oocyte from Chirostoma humboldtianum. The ovary is asynchronic, and as such, most phases of oocyte development are found in the same ovary. The complete process of oogenesis was divided in five stages: oogonium and folliculogenesis, primary growth, cortical alveoli and lipid inclusions, vitellogenesis, and maturation. The presence of big filaments, which appear at the end of primary growth, induces some common follicular adaptation. During primary growth, abundant ribosomes, the rough endoplasmic reticulum, and mitochondria are grouped in the cytoplasm. At the end of this stage, the Z1 layer of the chorion is developed, while microvilli start to be evident. In the cortical alveoli and lipid droplets phase, intense PAS positive vesicles, some of them containing nucleoid material, are observed in the peripheral cytoplasm and the lipid droplets take a more central position. In vitellogenesis, the proteic yolk accumulates in a centripetal way while the chorion is completely formed. During maturation, the germinal vesicle migrates to the animal pole, meiosis is restored, and there is nuclear breakdown. The oocyte increases its size and holds some oil droplets and a big fluid mass of yolk. On the outside, filaments completely surround the oocyte. Rev. Biol. Trop. 56 (3): 1371-1380. Epub 2008 September 30.

Histological seasonal changes in ovaries of Spotted tinamous (Nothura maculosa Tinamidae, Temminck, 1815) related to gonadotrope population / Cambios histológicos estacionales en ovarios de perdiz común (Nothura maculosa Tinamidae, Temminck, 1815)) y su relación con la población de gonadotropas

Nothura maculosa is a South American Tinamidae with a marked seasonal reproductive pattern. This work describes ovarian seasonal changes in this species related to gonadotrope (GTHs) population. Ovary and pituitary samples were collected monthly from adult birds during four annual periods. They were fixed in Bouin's solution and processed for light microscopy. The data of post-fixation gonadal weight were analysed using STATISTIX 4.0. Histological sections of the ovaries were stained with H/E, PAS and Goldner-Masson trichrome. Single and double immunostaining were applied on pituitary sections with anti-chicken-FSH and anti-chicken-LH antibodies. The samples were analysed in quarterly periods of the year, Pl: March-May (resting stage); P2: June-August (developing stage); P3: September-November (reproductive stage); P4: December-February (involutive stage). Ovary weight (ow) significantly varied among periods (p<0.001). During Pl, only primordial and previtellogenic follicles were observed, ow 0.09±0.01 g (n=25); during P2, developing follicles with signs of vitellogenesis were detected, ow 0.13+0.01 g (n=14); during P3, maximum follicular development was found, ow 0.9 +/- 0.15 g (n=39); P4 exhibited great variability in follicular stages, ow 0.18+0.18 g (n=19). Involutive atresia was observed in all the periods, while bursting atresia and post-ovulatory follicles were only characterized at P3 and P4. The GTHs containing few LH and FSH immunoreactive (ir) granules were predominant during P1-P2. The GTHs with LH ir granules were abundant in intermediate zone and caudal lobe in P3 and P4 while few cells contained both types of granules. The number of FSH cells was increased during P3 and P4. The histological ovarían changes were narrowly correlated with the variations in the gonadotrope population.

Nothura maculosa es un tinámido sudamericano que presenta marcada estacionalidad reproductiva. Este trabajo describe los cambios estacionales del ovario de esta especie, en relación con la población de gonadotropas (GTHs). Muestras de ovarios y pituitarias de ejemplares adultos fueron colectadas mensualmente durante cuatro años; se fijaron en solución de Bouin y procesadas para M.O. Los datos del peso gonadal posfijación fueron analizados usando STATISTIX 4.0. Los cortes de ovarios fueron coloreados con H/E, P.A.S. y Tricrómico de Goldner-Masson. En cortes de adenohipófisis se aplicó inmunocitoquímica simple y doble (sistema ABC, Vector Lab.), empleando anticuerpos anti-pollo FSH y anti-pollo LH. Las muestras se analizaron en períodos trimestrales de cada año (P): Pl: Marzo-Abril-Mayo (etapa de reposo), P2: Junio-Julio-Agosto (etapa de desarrollo), P3: Septiembre-Octubre-Noviembre (etapa reproductiva), P4: Diciembre-Enero-Febrero (etapa involutiva). El peso de los ovarios (PO) varió significativamente entre los periodos (p< 0.001). Durante Pl, sólo se observaron folículos primordiales y pre-vitelogénicos, PO 0.09 +/- 0.01 g (n=25); durante P2, se detectaron folículos en desarrollo con signos de vitelogénesis, PO 0.13+0.01 g (n=14); durante P3, se encontró máximo desarrollo folicular, PO 0.90+0.15 g (n=39); P4 exhibió gran variabilidad folicular, PO 0.18+0.18 g (n=19). La atresia involutiva se observó en todos los períodos, mientras que la atresia explosiva y los folículos postovulatorios caracterizaron a P3 y P4. Las GTHs conteniendo escasos granulos LH y FSH inmunoreactivos (ir) predominaron durante Pl y P2. Las GTHs con granulos LH¡> eran abundantes en la zona intermedia y en el lóbulo caudal en P3 y P4 mientras que escasas células contenían ambos tipos de granulos. El número de células FSH¡> se incrementó durante P3 y P4. Los cambios histológicos del ovario se correlacionaron estrechamente con las variaciones en la población de gonadotropas.