Decreased growth differentiation factor 9, bone morphogenetic protein 15, and forkhead box O3a expressions in the ovary via ulipristal acetate

SUMMARY OBJECTIVE: Folliculogenesis is a complex process involving various ovarian paracrine factors. During folliculogenesis, vitamin D3 and progesterone are significant for the proper development of follicles. This study aimed to investigate the effects of vitamin D3 and selective progesterone receptor modulator ulipristal acetate on ovarian paracrine factors. METHODS: In the study, 18 female Wistar-albino rats were randomly divided into three groups: control group (saline administration, n=6), vitamin D3 group (300 ng/day vitamin D3 oral administration, n=6), and UPA group (3 mg/kg/day ulipristal acetate oral administration, n=6). Ovarian tissue was analyzed by histochemistry and immunohistochemistry. For quantification of immunohistochemistry, the mean intensities of growth differentiation factor 9, bone morphogenetic protein 15, and forkhead box O3a expressions were measured by Image J and MATLAB. Blood samples were collected for the analysis of serum anti-Müllerian hormone levels by ELISA. RESULTS: Atretic follicles and hemorrhagic cystic structures were observed in the UPA group. After immunohistochemistry via folliculogenesis assessment markers, growth differentiation factor 9, bone morphogenetic protein 15, and cytoplasmic forkhead box O3a expressions decreased in the UPA group (p<0.05). Anti-Müllerian hormone level did not differ significantly between the experimental groups (p>0.05). CONCLUSION: Ulipristal acetate negatively affects folliculogenesis via ovarian paracrine factors. The recommended dietary vitamin D3 supplementation in healthy cases did not cause a significant change.

Manipulação de oócitos inclusos em folículos ovarianos pré-antrais e a importância dos antioxidantes ­ revisão de literatura / Manipulation of oocytes included in preantral ovarian follicles and the importance of antioxidants ­ literature review / Manipulación de ovocitos incluidos en folículos ováricos preantrales e importancia de los antioxidantes - revisión de la literatura

A manipulação de oócitos inclusos em folículos ovarianos pré-antrais (MOIFOPA) vem sendo estudada pensando na perspectiva futura de aplicação direta na reprodução humana, principalmente para mulheres que sofrem de doenças ou que precisam passar por tratamentos que interferem na função ovariana. Nesse contexto, o objetivo deste trabalho é revisar aspectos relacionados com a biotécnica de MOIFOPA e a importância dos antioxidantes no cultivo in vitro de folículos pré-antrais. Foi realizada uma pesquisa na base de dados PubMed, buscando artigos sobre a biotécnica, principalmente relacionados com a necessidade do uso de antioxidantes no cultivo. A grande maioria dos estudos sobre a biotécnica utilizam como modelo experimental os folículos ovarianos de diferentes espécies de animais. A MOIFOPA compreende o isolamento e o resgate de folículos ovarianos pré-antrais provenientes de ovários, seguido da conservação através da técnica de resfriamento ou congelação e o cultivo folicular in vitro, a fim de promover o crescimento, a maturação e a fecundação in vitro (FIV) dos oócitos inclusos nesses folículos, maximizando o potencial reprodutivo feminino e diminuindo a atresia folicular que acontece in vivo. Um aspecto que pode interferir no sucesso do cultivo in vitro de folículos ovarianos pré-antrais é a produção em excesso de espécies reativas de oxigênio (EROs). Os ácidos ascórbico e alfa lipóico vem demonstrando resultados interessantes para reduzir os efeitos que as EROs causam sobre os folículos ovarianos pré-antrais cultivados in vitro.

The manipulation of oocytes included in preantral ovarian follicles (MOEPF) has been studied considering the future perspective of direct application in human reproduction, especially for women who suffer from diseases or who need to undergo treatments that interfere with ovarian function. In this context, the objective of this paper is to review aspects related to the biotechnology of MOIFOPA and the importance of antioxidants. A search was carried out in the PubMed database, searching for articles on biotechnology, mainly related to the need to use antioxidants in cultivation. The vast majority of studies on biotechnology use ovarian follicles from different species of animals as an experimental model. MOIFOPA comprises the isolation and rescue of preantral ovarian follicles from ovaries, followed by conservation through the cooling or freezing technique and in vitro follicular cultivation, in order to promote growth, maturation and in vitro fertilization ( IVF) of the oocytes included in these follicles, maximizing the female reproductive potential and decreasing the follicular atresia that occurs in vivo. One aspect that may interfere with the success of in vitro culture of preantral ovarian follicles is the excess production of reactive oxygen species (ROS). Ascorbic and alpha lipoic acids have shown interesting results in reducing the effects that ROS cause on in vitro cultured preantral ovarian follicles.

manipulación de ovocitos incluidos en folículos ováricos preantrales (MOIFOPA) se ha estudiado con la perspectiva futura de su aplicación directa en la reproducción humana, especialmente en mujeres que padecen enfermedades o que necesitan someterse a tratamientos que interfieren en la función ovárica. En este contexto, el objetivo de este trabajo es revisar los aspectos relacionados con la biotécnica de MOIFOPA y la importancia de los antioxidantes en el cultivo in vitro de los folículos pré-antrais. Se realizó una investigación en la base de datos PubMed, buscando artículos sobre la biotecnología, principalmente relacionados con la necesidad del uso de antioxidantes en el cultivo. La mayoría de los estudios sobre biotecnología utilizan como modelo experimental los folículos ováricos de diferentes especies de animales. El MOIFOPA incluye el aislamiento y rescate de los folículos ováricos preantrales de los ovarios, seguido de su conservación mediante la técnica de enfriamiento o congelación y el cultivo folicular in vitro, con el fin de promover el crecimiento, la maduración y la fecundación in vitro (FIV) de los ovocitos incluidos en estos folículos, maximizando el potencial reproductivo femenino y disminuyendo la atresia folicular que se produce in vivo. Un aspecto que puede interferir en el éxito del cultivo in vitro de folículos ováricos preantrales es la producción excesiva de especies reactivas de oxígeno (ROS). El ácido ascórbico y el ácido alfa lipoico han mostrado resultados interesantes para reducir los efectos que causan las ERO en los folículos ováricos preantrales cultivados in vitro.

Electrophysiological responses to different follicle-stimulating hormone isoforms on human cumulus oophorus cells: preliminary results / Respostas eletrofisiológicas a diferentes isoformas de FSH em células humanas do cumulus oophorus: resultados preliminares

Abstract Objective The aim of the present study was to provide a better understanding of the specific action of two follicle-stimulating hormone (FSH) isoforms (β-follitropin and sheep FSH) on the membrane potential of human cumulus cells. Methods Electrophysiological data were associated with the characteristics of the patient, such as age and cause of infertility. The membrane potential of cumulus cells was recorded with borosilicate microelectrodes filled with KCl (3 M) with tip resistance of 15 to 25 MΩ. Sheep FSH and β-follitropin were topically administered onto the cells after stabilization of the resting potential for at least 5 minutes. Results In cumulus cells, the mean resting membrane potential was - 34.02 ± 2.04 mV (n = 14). The mean membrane resistance was 16.5 ± 1.8 MΩ (n = 14). Sheep FSH (4 mUI/mL) and β-follitropin (4 mUI/mL) produced depolarization in the membrane potential 180 and 120 seconds after the administration of the hormone, respectively. Conclusion Both FSH isoforms induced similar depolarization patterns, but β-follitropin presented a faster response. A better understanding of the differences of the effects of FSH isoforms on cell membrane potential shall contribute to improve the use of gonadotrophins in fertility treatments.

Resumo Objetivo O objetivo do presente estudo foi fornecer uma melhor compreensão da ação específica de duas isoformas de hormônio folículo estimulante (FSH, sigla em inglês) (β-folitropina e FSH ovino) no potencial de membrana de células do cumulus oophorus humanas. Métodos Dados eletrofisiológicos foram associados às características da paciente, como idade e causa da infertilidade. O potencial de membrana das células do cumulus foi registrado com microeletrodos de borossilicato preenchidos com KCl (3 M) com uma resistência de 15 a 25 MΩ. O FSH ovino e a β-folitropina foram administrados topicamente nas células após a estabilização do potencial de repouso durante pelo menos 5 minutos. Resultados Nas células do cumulus, o potencial médio de membrana em repouso foi de -34,02 ± 2,04 mV (n = 14). A resistência média da membrana foi de 16,5 ± 1,8 MΩ (n = 14). O FSH ovino (4 mUI/mL) e a β-folitropina (4 mUI/mL) produziram despolarização no potencial de membrana 180 e 120 segundos após a aplicação do hormônio, respectivamente. Conclusão Ambas as isoformas de FSH induzem padrões de despolarização semelhantes, mas a β-folitropina apresentou uma resposta mais rápida. Uma melhor compreensão das diferenças dos efeitos das isoformas do FSH no potencial da membrana celular contribuirá para aprimorar o uso das gonadotrofinas no estímulo ovariano controlado e em protocolos de maturação oocitária in vitro.

Reports on in vivo and in vitro contribution of medicinal plants to improve the female reproductive function / Relatos sobre a contribuição in vivo e in vitro de plantas medicinais na melhora da função reprodutiva feminina

Medicinal plants are known as a prolific source of secondary metabolites which have important function both in vivo and in vitro during the ovarian folliculogenesis and steroidogenesis in many animal species. Some secondary metabolites can act as antioxidants generally through their ability to scavenge reactive oxygen species (ROS) or can regulate ovarian hormonal production. In general, these properties are responsible for the medicinal functions to treat woman infertility disorder. Some plants are constituted of biological actives substances which have been used to treat reproductive dysfunction. However, until recently, little was known about the implication of plants and/or their secondary metabolites on in vitro folliculogenesis and steroidogenesis. With the development of the technology, there is an increase implication of those substances in assisted reproductive technology (ART). The present review highlights some medicinal plants used in the treatment of woman disorders related to infertility. In addition, it provides an in vivo and in vitro overview of herbs and their active compounds with claims for improvement of ovarian activity thus showing their implication in female reproductive health care.(AU)

Sabe-se que as plantas medicinais são uma fonte abundante de metabólitos secundários que têm função importante tanto in vivo quanto in vitro durante a foliculogênese e a esteroidogênese ovarianas em muitas espécies animais. Alguns metabólitos secundários podem atuar como antioxidantes, geralmente através de sua capacidade de eliminar espécies reativas de oxigênio (ROS) ou podem regular a produção hormonal ovariana. Em geral, essas propriedades são responsáveis pelas funções medicinais usadas para tratar distúrbios da infertilidade feminina. Algumas plantas contêm substâncias biológicas ativas que têm sido utilizadas para tratar a disfunção reprodutiva. No entanto, até recentemente, pouco se sabia sobre o efeito das plantas e/ou seus metabólitos secundários na foliculogênese e na esteroidogênese in vitro. Com o desenvolvimento da tecnologia, há uma implicação crescente dessas substâncias na tecnologia de reprodução assistida (TRA). A presente revisão destaca algumas plantas medicinais utilizadas no tratamento de distúrbios femininos relacionados à infertilidade. Além disso, fornece uma visão in vivo e in vitro de ervas e seus compostos ativos com alegações de melhora da atividade ovariana, mostrando assim seu envolvimento nos cuidados de saúde reprodutiva feminina.(AU)

Morphologic and morphometric analysis of the ovary of black bengal goat (Capra hircus) / Análisis morfológico y morfométrico del ovario de la cabra black bengal (Capra hircus)

Ovary plays the vital role in the reproductive biology and biotechnology of female animals. With the aim to study the ovarian morphometry of Black Bengal goat, both right and left ovaries were collected from the slaughter houses of different Thanas under Mymensingh district. For each of the specimens, gross parameters such as weight, length and width were recorded. Then they were processed and stained with H & E for histomorphometry. Our study revealed that the right ovary (0.53±0.02 g) was heavier than the left (0.52±0.02 g). The length of the right ovary (1.26±0.04 cm) was lower than the left (1.28±0.02 cm) but the width of the right (0.94±0.02 cm) was greater than the left (0.90±0.03 cm). The diameter of ovarian follicles in the cortex was measured as primordial 39.6±6.61 µm, primary single layer 54.0±4.06 µm, primary multi layer 147.6±11.04 µm, secondary with C-shaped antrum 449.5±75.71 µm and tertiary follicle of ovary 1.3±0.20 mm. In the graffian follicle, the thickness of granulosa cell layer was 79.2±11.04 µm, theca interna 75.76±6.82 µm, theca externa 130.07±12.53 µm and the oocyte diameter was 109.8±5.75 µm. These results will be helpful to manipulate ovarian functions in small ruminants.

El ovario desempeña un papel fundamental en la biología de la reproducción y la biotecnología de las hembras. Con el objetivo de estudiar la morfometría de ovario de la cabra Black Bengal se obtuvieron ovarios de ambos lados de diferentes mataderos del distrito de Mymensingh. Para cada uno de los especímenes, se registraron los parámetros de peso, longitud y ancho. Luego fueron procesados y teñidos con H & E para histomorfometría. El ovario derecho (0,53±0,02 g) fue más pesado que el izquierdo (0,52±0,02 g). La longitud del ovario derecho (1,26±0,04 cm) fue inferior al izquierdo (1,28±0,02 cm), pero el ancho del derecho (0,94±0,02 cm) fue mayor que el izquierdo (0,90±0,03 cm). El diámetro de los folículos ováricos en la corteza se midió como primordial 39,6±6,61 µm, de una capa primaria 54,0±4,06 µm, de múltiples capas primarias 147,6±11,04 µm, secundarias con antro en forma de C 449,5±75,71 µm y folículo terciario del ovario 1,3±0,20 mm. En el folículo terciario del ovario, el espesor de la capa de células de la granulosa fue 79,2±11,04 µm, teca interna 75,76±6,82 µm, teca externa 130,07±12,53 µm y el diámetro de los ovocitos fue 109,8±5,75 µm. Estos resultados serán útiles para manipular las funciones de ovario en los pequeños rumiantes.

Follicular dynamics, corpus luteum growth and regression in multiparous buffalo cows and buffalo heifers / Dinámica folicular, crecimiento y regresión del cuerpo lúteo en búfalas multíparas y novillas

Objective. Characterize the follicular dynamics and luteal growth and regression pattern of multiparous (MB) and heifer (BH) Murrah buffaloes in Colombian tropical conditions. Material and methods. Ten MB and ten BH were synchronized with a progesterone-releasing intravaginal device. No artificial insemination was performed during the estrous and daily ultrasound examinations were performed 15 days later to determine the number and diameter of the structures present in both ovaries in the subsequent natural estrous cycle. The Student's T test was used to evaluate differences between MB and BH. All data are presented as mean ± standard deviation. Results. The length of the estrous cycle was 22.00±4.50 days for MB and 22.00±2.70 days for BH. Follicular growth occurs in one (n=1; 5.89%), two (n=14; 82.35%) or three waves (n=2; 11.76%). The first wave initiated the day after ovulation with the recruitment of 8.33±2.06 and 10.00±2.72 follicles in MB and BH, while the second wave started on day 11.00±2.00 and 10.50±2.82, presenting 8.37±2.26 and 8.00±1.51 follicles. The third wave began on day 16.21±3.10 showing 6.50±1.70 follicles, only BM had three waves. The maximum luteal diameter was 19.58±4.16 mm and 17.74±3.32 mm respectively. There were no significant differences between the groups for these variables. Conclusions. These results show that the follicular development in buffaloes occurs in waves, where two waves is the most common pattern, as previously reported by other authors.

Objetivo. Caracterizar la dinámica folicular y el patrón de crecimiento y regresión del cuerpo lúteo de búfalas multíparas (BM) y búfalas novillas (BN) de la raza Murrah en condiciones del trópico colombiano. Materiales y métodos. Diez BM y diez BN fueron sincronizadas con dispositivo intravaginal de liberación de progesterona. No se realizó la inseminación artificial al momento del celo y 15 días después se inició el seguimiento ultrasonográfico para determinar el número y diámetro de las estructuras presentes en ambos ovarios en el subsecuente ciclo estral natural. Las diferencias entre BM y BN se evaluaron con pruebas T. Los datos se presentan como media ± desviación estándar. Resultados. La duración del ciclo estral fue de 22.00±4.50 y 22.00±2.70 días en BM y BN. El crecimiento folicular ocurrió en una (n=1; 5.89%), dos (n=14; 82.35%) o tres (n=2; 11.76%) ondas. La primera onda inicio el día siguiente a la ovulación con el reclutamiento de 8.33±2.06 y 10.00±2.72 folículos en BM y BN, mientras que la segunda onda inicio el día 11.00±2.00 y 10.50±2.82 con 8.37±2.26 y 8.00±1.51 folículos. La tercera onda inicio el día 16.21±3.10 con 6.50±1.70 folículos, sólo BM presentaron tres ondas. El diámetro máximo luteal fue de 19.58±4.16 mm y 17.74±3.32 mm. No se observaron diferencias significativas entre los grupos para estas variables. Conclusiones. Los resultados muestran que el desarrollo folicular de las búfalas se dio en ondas, siendo dos ondas el patrón más común, similar a lo reportado por otros autores.

Factores que afectan la preñez en vacas Brahman sometidas a inseminación artificial a tiempo fijo / Factors affecting pregnancy in Brahman cows submitted to fixed-time artificial insemination

Objetivo. Evaluar la relación entre la condición corporal (CC), la manifestación de estro, las concentraciones plasmáticas de estradiol, el diámetro del folículo mayor y la preñez, en vacas Bos indicus amamantando, sometidas a un protocolo de inseminación artificial a tiempo fijo (IATF). Materiales y métodos. Treinta y ocho vacas Brahman recibieron un dispositivo intravaginal de progesterona + benzoato de estradiol en el día 0; retiro del dispositivo + dinoprost + gonadotropina coriónica equina en el día 9 y la IATF en el día 12. La preñez fue diagnosticada 90 días después de la IATF. Resultados. No hubo efecto de la CC sobre la manifestación de estro, las concentraciones plasmáticas de estradiol o el diámetro del folículo mayor (p>0.05). El diámetro del folículo mayor no influenció las concentraciones plasmáticas de estradiol (p>0.05). El diámetro del folículo mayor y las concentraciones plasmáticas de estradiol no influenciaron la manifestación de estro (p>0.05). La tasa de preñez por IATF no fue afectada (p>0.05) por la CC, la manifestación de estro o las concentraciones plasmáticas de estradiol. Hubo una tendencia (p=0.10) de una relación directa entre el diámetro del folículo mayor y la tasa de preñez por IATF. Conclusiones. La CC, las concentraciones plasmáticas de estradiol y la manifestación de estro no influenciaron la tasa de preñez por IATF en vacas Brahman sometidas a un protocolo de sincronización basado en progesterona. Sin embargo, el diámetro del folículo mayor presentó tendencia al impacto positivo sobre la tasa de preñez por IATF.

Objective. To examine the relationship between body condition score (BCS), the occurrence of estrous, estradiol plasma concentrations, diameter of largest ovarian follicle (LF) and pregnancy rate in suckled Bos indicus cows submitted to a fixed-time artificial insemination (FTAI) protocol. Material and methods. Thirty eight Brahman cows received an intravaginal device of progesterone + estradiol benzoate on Day 0, device withdrawal + dinoprost + equine chorionic gonadotropin on Day 9, and FTAI on Day 12. Pregnancy was diagnosed 90 days after FTAI. Results. There was no effect of BCS on the occurrence of estrous, estradiol plasma concentrations or diameter of the LF (p>0.05). The diameter of LF did not influence (p>0.05) estradiol plasma concentrations. The diameter of the LF and estradiol plasma concentrations did not influence the occurrence of estrous (p>0.05). Pregnancy rate per FTAI was not affected (p>0.05) by BCS, the occurrence of estrous or plasma concentrations of estradiol. There was a tendency (p=0.10) of a direct relationship between diameter of the LF and pregnancy rate FTAI. Conclusions. BCS, plasma concentrations of estradiol and occurrence of estrous did not influence pregnancy rate per FTAI in Brahman cows submitted to progesterone-based protocol. However, the diameter of LF had a tendency to positive impact on pregnancy rate per FTAI.

Dinámica folicular ovárica durante el ciclo estral en vacas brahman / Ovarian Follicular Dynamics During the Estrous Cycle in Brahman Cows

Con el objetivo de caracterizar la dinámica folicular durante el ciclo estral (CE) en vacas Brahman, fueron evaluados 12 CE’s con duración promedio de 21,3 ± 0,7 d (rango=19-24 d). Setenta y cinco porciento (9/12) de los CE’s presentaron dos ondas de crecimiento folicular (CF), emergiendo los días 1,6 ± 0,4 y 8,4 ± 0,4 del ciclo, respectivamente. En CE’s de una sola onda (2/12), la emergencia ocurrió el día 14,5 ± 0,7; mientras en el CE de tres ondas (1/12), las mismas emergieron los días 2, 10 y 17. Durante el CE ocurrieron cambios en las clases foliculares (Clase 1 [C1]: ≤5 mm; Clase 2 [C2]: 6-9 mm; Clase 3 [C3]: ≥10 mm). Disminuyó el número de folículos C1 mientras aumentó el número de folículos C2 durante el reclutamiento, y descendió el número de folículos C2 mientras aumentó el número de folículos C3 durante la selección. La tasa de crecimiento (TC) fue mayor (P<0,05) para el folículo dominante (FD1= 0,78 ± 0,1 mm/d; FD2=0,40 ± 0,1 mm/d), que en el folículo subordinado (FS1= -0,01 ± 0,1 mm/d; FS2= 0,22 ± 0,1 mm/d). No hubo diferencia en el máximo diámetro alcanzado por el FD (FD1= 12,2 ± 0,4 mm; FD2= 12,7 ± 0,4 mm). El diámetro del cuerpo lúteo (CL) aumentó (fase de crecimiento) hasta alcanzar su máximo tamaño (fase de meseta), disminuyendo hasta sudesaparición (fase de regresión). Los niveles de P4 mostraron correlación positiva con el diámetro del CL (r=0,54; P<0,01), aumentando los niveles de P4 durante la fase de crecimiento del CL y disminuyendo durante su regresión, con poca variación durante la fase de meseta. Estos resultados permiten señalar que la dinámica folicular en vacas Brahman presenta los mismos patrones observados en ganado Bos indicus, previamente reportados.

To characterize the follicular dynamics during the estrous cycle (EC) in Brahman cows, 12 ECs with a mean duration of 21.3 ± 0.7 d (range= 19-24 d), were evaluated. Seventy five percent (9/12) of the EC’s had two waves of follicular growth (FG; 20.6+0.5 d), emerging on days 1.6+0.4 and 8.4+0.4, respectively. In EC’s of one wave of FG (2/12), emergence occurred on day 14.5+0.7; whereas in EC’s with three waves (1/12), waves emerged on days 2, 10 and 17, respectively. During the EC, the number of follicles (Class 1 [C1]= ≤ 5 mm, Class 2 [C2]= 6-9 mm; Class 3 [C3]= ≥ 10 mm) changed, showing a decrease in C1 follicles and an increase in C2 follicles, during the recruitment phase, and a decrease in C2 follicles during the selection phase, with an increase in C3 follicles. The growth rate of the dominant follicle (DF; DF1=0.78 ± 0.1 mm/d; FD2= 0.40 ± 0.1 mm/d) was greater (P<0.05) than the growth rate for subordinate follicles (SF1=-0.01 ± 0.1 mm/d; SF2=0.22 ± 0.1 mm/d). There was no difference in size between DF1 and DF2 (12.2±0.4 vs. 12.7 ± 0.4 mm). From the beginning the diameter of the corpus luteum (CL) increased (growth phase) until it reached the maximum size (plateau phase), to decrease due to its regression. Plasma progesterone (P4) levels showed a positive correlation (r=0.54; P< 0.01) with the diameter of the CL. These results allowed to conclude that the follicular dynamics during the EC in Brahman cows had the same patterns observed in Bos indicus and Bos taurus cattle, previously reported.

Ultrastructure and morphometry of ovarian follicles in the armadillo Chaetophractus villosus (Mammalia, Dasypodidae)

Ultrastructural and morphometric changes in oocyte and surrounding granulosa cells during armadillo follicular growth are described. Primordial, intermediary, early and late primary, secondary, tertiary and preovulatory Graafian follicles were found. From primordial to Graafian follicle, granulosa cells increase in height, become multilayered while fluid-filled spaces arise among them in the tertiary follicle stage. As the follicle expands the oocyte is located eccentrically and grows in size. From primordial to secondary stage organelles develop in number and complexity and many vesicles start arising in the cytoplasm at the secondary stage. From tertiary follicle stage the number of mitochondria decreases. In follicular cells, secreting-related organelles expand, but mitochondrial number decreases according to the follicle expansion. Intercellular junctions between both cell types increase. The morphometrical study shows a biphasic pattern of oocyte and follicle growth. The morphological baseline here provided will be essential for further comprehension of the reproductive biology in armadillos.

São descritas a ultra-estrutura e as mudanças morfométricas nos ovócitos e nas células da granulosa durante o crescimento folicular do tatu. Foram encontrados folículos primordiais, intermediários, primários (iniciais e finais), secundários, terciários e folículos de Graff pré-ovulatórios. Desde os folículos primordiais até os folículos de Graff, as células da granulosa aumentam de tamanho, dispõem-se em múltiplas camadas e, no folículo terciário, apresentam espaços cheios de líquido entre elas. Quando o folículo se expande, o ovócito localiza-se exentricamente e aumenta de tamanho. Nos folículos primários e secundários, as organelas desenvolvem-se aumentando em número e complexidade e, no folículo secundário, várias vesículas desenvolvem-se no citoplasma. A partir do folículo terciário, o número de mitocôndrias diminui. Nas células foliculares, as organelas relacionadas à secreção se expandem, mas o número de mitocôndrias diminui com a expansão do folículo. As uniões intercelulares entre ambos os tipos de células aumentam. Os estudos morfométricos mostram um padrão bifásico de crescimento do ovócito e do folículo. A informação morfológica deste trabalho pode ser de grande importância no entendimento da biologia reprodutiva dos tatus.