Bilateral canine seminoma with ocular metastasis: histochemical and immunohistochemical characterization

A 10-year old male dog was examined due to a buphthalmia in the left eye and a nodule in the two testicles. Due to the limited resources of the owner and loss of visual acuity of the patient, the enucleation and castration were chosen as treatment. Microscopic analysis of the testicular tissue revealed neoplastic germ cells. Morphologically, neoplastic cells were characterized by distinct cell borders, scarce and eosinophilic cytoplasm, large round nucleus, with thick chromatin and a prominent nucleolus. Binucleated and multinucleated neoplastic cells were also frequently observed. In 10 high powerfields (400x), 62 typical and atypical mitosis were counted. Similar neoplastic cells were identified within the vessels of theretina, sclera and in the sub-epithelial conjunctive tissue of the eyelid. The neoplastic cells observed in the testicle and in the eye were positive for PAS. By immunochemistry technique was identified an intense immunostaining of the neoplastic cells for Vimentin and Ki-67 in both testicular and ocular tissue. While, discrete immunoreactivity was identified to c-KIT from the neoplastic cells in both organs. Based on morphological, histochemical and immunohistochemical analysis, it was possible to characterize the ocular lesion as seminoma metastasis.

Bilateral canine seminoma with ocular metastasis: histochemical and immunohistochemical characterization

A 10-year old male dog was examined due to a buphthalmia in the left eye and a nodule in the two testicles. Due to the limited resources of the owner and loss of visual acuity of the patient, the enucleation and castration were chosen as treatment. Microscopic analysis of the testicular tissue revealed neoplastic germ cells. Morphologically, neoplastic cells were characterized by distinct cell borders, scarce and eosinophilic cytoplasm, large round nucleus, with thick chromatin and a prominent nucleolus. Binucleated and multinucleated neoplastic cells were also frequently observed. In 10 high powerfields (400x), 62 typical and atypical mitosis were counted. Similar neoplastic cells were identified within the vessels of theretina, sclera and in the sub-epithelial conjunctive tissue of the eyelid. The neoplastic cells observed in the testicle and in the eye were positive for PAS. By immunochemistry technique was identified an intense immunostaining of the neoplastic cells for Vimentin and Ki-67 in both testicular and ocular tissue. While, discrete immunoreactivity was identified to c-KIT from the neoplastic cells in both organs. Based on morphological, histochemical and immunohistochemical analysis, it was possible to characterize the ocular lesion as seminoma metastasis.(AU)

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.(AU)

Isolation and characterization of the human ovarian cell population for transplantation into an artificial ovary

To support survival and growth of follicles, the transplantable artificial ovary should mimic the original organ, offering a physical (3D matrix) and biological support (cells). In order to replicate the ovarian cell populations, the aim of this study is to assess the proportions of stromal and endothelial cells in the ovarian cortex. To this end, ovarian biopsies were obtained from six women (mean age: 49 years). The epithelial layer and medulla were carefully removed. The cortex was finely minced and enzymatically digested and the isolated cells were fixed. For cell characterization, immunostaining for CD31 (for endothelial cells) and inhibin-α (for granulosa cells) was performed. Positive cells in each staining were counted and the proportion of the different cell populations was estimated from the total number of isolated cells. Since there is no specific marker for ovarian stromal cells, we estimated the proportion of these cells by performing a vimentin immunostaining and subtracting the proportions of CD31- and inhibin-α-positive cells. Immunostaining showed that 84% of isolated cells were vimentin-positive. From this pool, 3% were endothelial cells and 1% granulosa cells. Consequently, the population of ovarian stromal cells was 80%. In conclusion, our findings show that stromal cells represent the larger population of cells in the human ovarian cortex. While this ensures follicle survival and development in a normal ovary, we believe that the low proportion of endothelial cells could have a negative impact on the angiogenesis in the artificial ovary after the first days of transplantation.

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.

Isolation and characterization of the human ovarian cell population for transplantation into an artificial ovary

To support survival and growth of follicles, the transplantable artificial ovary should mimic the original organ, offering a physical (3D matrix) and biological support (cells). In order to replicate the ovarian cell populations, the aim of this study is to assess the proportions of stromal and endothelial cells in the ovarian cortex. To this end, ovarian biopsies were obtained from six women (mean age: 49 years). The epithelial layer and medulla were carefully removed. The cortex was finely minced and enzymatically digested and the isolated cells were fixed. For cell characterization, immunostaining for CD31 (for endothelial cells) and inhibin-α (for granulosa cells) was performed. Positive cells in each staining were counted and the proportion of the different cell populations was estimated from the total number of isolated cells. Since there is no specific marker for ovarian stromal cells, we estimated the proportion of these cells by performing a vimentin immunostaining and subtracting the proportions of CD31- and inhibin-α-positive cells. Immunostaining showed that 84% of isolated cells were vimentin-positive. From this pool, 3% were endothelial cells and 1% granulosa cells. Consequently, the population of ovarian stromal cells was 80%. In conclusion, our findings show that stromal cells represent the larger population of cells in the human ovarian cortex. While this ensures follicle survival and development in a normal ovary, we believe that the low proportion of endothelial cells could have a negative impact on the angiogenesis in the artificial ovary after the first days of transplantation.(AU)

Transforming growth factor-beta family members are regulated during induced luteolysis in cattle

The transforming growth factors beta (TGFβ) are local factors produced by ovarian cells which, after binding to their receptors, regulate follicular deviation and ovulation. However, their regulation and function during corpus luteum (CL) regression has been poorly investigated. The present study evaluated the mRNA regulation of some TGFβ family ligands and their receptors in the bovine CL during induced luteolysis in vivo. On day 10 of the estrous cycle, cows received an injection of prostaglandin F2α (PGF) and luteal samples were obtained from separate groups of cows (n= 4-5 cows per time-point) at 0, 2, 12, 24 or 48 h after treatment. Since TGF beta family comprises more than 30 ligands, we focused in some candidates genes such as activin receptors (ACVR-1A, -1B, -2A, -2B) AMH, AMHR2, BMPs (BMP-1, -2, -3, -4, -6 and -7), BMP receptors (BMPR-1A, -1B and -2), inhibin subunits (INH-A, -BA, -BB) and betaglycan (TGFBR3). The mRNA levels of BMP4, BMP6 and INHBA were higher at 2 h after PGF administration (P<0.05) in comparison to 0 h. The relative mRNA abundance of BMP1, BMP2, BMP3, BMP4, BMP6, ACVR1B, INHBA and INHBB was upregulated up to 12 h post PGF (P<0.05). On the other hand, TGFBR3 mRNA that codes for a reservoir of ligands that bind to TGF-beta receptors, was lower at 48 h. In conclusion, findings from this study demonstrated that genes encoding several TGFβ family members are expressed in a time-specific manner after PGF administration.(AU)

Transforming growth factor-beta family members are regulated during induced luteolysis in cattle

The transforming growth factors beta (TGFβ) are local factors produced by ovarian cells which, after binding to their receptors, regulate follicular deviation and ovulation. However, their regulation and function during corpus luteum (CL) regression has been poorly investigated. The present study evaluated the mRNA regulation of some TGFβ family ligands and their receptors in the bovine CL during induced luteolysis in vivo. On day 10 of the estrous cycle, cows received an injection of prostaglandin F2α (PGF) and luteal samples were obtained from separate groups of cows (n= 4-5 cows per time-point) at 0, 2, 12, 24 or 48 h after treatment. Since TGF beta family comprises more than 30 ligands, we focused in some candidates genes such as activin receptors (ACVR-1A, -1B, -2A, -2B) AMH, AMHR2, BMPs (BMP-1, -2, -3, -4, -6 and -7), BMP receptors (BMPR-1A, -1B and -2), inhibin subunits (INH-A, -BA, -BB) and betaglycan (TGFBR3). The mRNA levels of BMP4, BMP6 and INHBA were higher at 2 h after PGF administration (P<0.05) in comparison to 0 h. The relative mRNA abundance of BMP1, BMP2, BMP3, BMP4, BMP6, ACVR1B, INHBA and INHBB was upregulated up to 12 h post PGF (P<0.05). On the other hand, TGFBR3 mRNA that codes for a reservoir of ligands that bind to TGF-beta receptors, was lower at 48 h. In conclusion, findings from this study demonstrated that genes encoding several TGFβ family members are expressed in a time-specific manner after PGF administration.

Neoplasias em equinos, asininos e muares

Os equinos, asininos e muares têm importante participação na economia e as neoplasias nestas espécies são importantes causas de perdas econômicas. As neoplasias em equídeos são diagnosticadas através de necropsias e biópsias, sendo o principal sistema afetado o tegumentar, principalmente em decorrência do sarcoide. No entanto é importante incluir neoplasias de outros sistemas, inclusive em órgãos internos como importantes diagnósticos diferenciais que causam debilidade e podem até levar a morte. Esta dissertação foi elaborada em dois capítulos, compostos por dois artigos originais. O primeiro a ser submetido à revista Pesquisa Veterinária Brasileira relata a frequência de tumores diagnosticados em nove sistemas primários de equídeos, descrevendo aspectos epidemiológicos, clínicos e patológicos. Foram diagnosticadas 186 neoplasias em 167 equídeos. O sistema mais afetado foi o tegumentar seguindo do reprodutor masculino e feminino, ocular/periocular, respiratório, digestório, hepático, nervoso e hematopoiético. A neoplasia mais diagnosticada foi o sarcoide, seguida do carcinoma de células escamosas e melanoma. Metástases ocorreram em 14 casos e os sinais clínicos foram variáveis de acordo com a localização anatômica. O segundo artigo será submetido à Journal of Equine Veterinary Science e descreve-se três casos de tumor das células da granulosa diagnosticados em éguas, abordando os aspectos epidemiológicos, clínicos e patológicos. As éguas apresentavam alterações comportamentais e reprodutivos como infertilidade, repetição de cio (ninfomania), comportamento de garanhão (virilidade), indiferença ao garanhão e ausência de sinais de estro. As neoplasias foram visualizados através da ultrassonografia e removidos cirurgicamente pela ovariectomia e enviados para exames histopatológicos. Macroscopicamente as massas variavam de 9 a 20 cm de diâmetro, eram firmes e bem delimitadas com superfícies lisas e ao corte observou-se múltiplas cavidades císticas circundadas por trabéculas de tecido amarelo-esbranquiçado e firmes. Microscopicamente observou-se massas multilobuladas, bem delimitadas, compostas por células dispostas em ninhos semelhantes a folículos abortados ou formando cistos, apoiadas em moderado estroma fibrocolagenoso. Pseudorosetas foram observadas. Na neoplasia da égua 2, observou-se formações semelhantes a túbulos de sertoli e células semelhantes a células de Leydig. Na imuno-histoquímica, houve marcação pela inibina e vimetina, de células da granulosa maduras e neoplásicas que compunham os ninhos e revestiam cistos. Após a cirurgia, a égua 1 não exibiu mais as alterações comportamentais, apresentou cio duas vezes, ficou prenhe e abortou. A égua 3 também ficou prenhe e pariu a termo. Não há informações referentes a éguas 2 após a cirurgia. Foi possível constatar com esta dissertação que embora a maioria das neoplasias diagnosticadas em equídeos no Laboratório de Patologia Animal do Hospital Veterinário Universitário da UFCG, envolvam órgãos relacionados ao tegumento, neoplasias que acometem órgãos internos ocorrem esporadicamente e é importante incluí-las como diagnósticos diferenciais.

Horses, donkeys and mules play an important role in the economy and neoplasms in these species are important causes of economic losses. Neoplasms in equines are diagnosed through necropsies and biopsies, with the integumentary system being the main affected system, mainly due to the sarcoid. However, it is important to include neoplasms from other systems, including internal organs as important differential diagnoses that cause debility and can even lead to death. This dissertation was elaborated in two chapters, composed by two original articles. The first to be submitted to the journal Pesquisa Veterinária Brasileira reports the frequency of tumors diagnosed in nine primary equine systems, describing epidemiological, clinical and pathological aspects. 186 neoplasms were diagnosed in 167 horses. The most affected system was the integumentary, followed by the male and female, ocular/periocular, respiratory, digestive, hepatic, nervous and hematopoietic systems. The most diagnosed neoplasm was sarcoid, followed by squamous cell carcinoma and melanoma. Metastases occurred in 14 cases and clinical signs varied according to anatomical location. The second article will be submitted to the Journal of Equine Veterinary Science and describes three cases of granulosa cell tumor diagnosed in mares, addressing the epidemiological, clinical and pathological aspects. The mares showed behavioral and reproductive changes such as infertility, repetition of heat (nymphomania), stallion behavior (virility), indifference to the stallion and absence of signs of estrus. The neoplasms were visualized through ultrasonography and surgically removed by ovariectomy and sent for histopathological exams. Macroscopically, as masses ranged from 9 to 20 cm in diameter, they were firm and well delimited with smooth surfaces and, when cut, multiple cystic cavities surrounded by firm yellow-white tissue trabeculae were observed. Microscopically, well-delimited, multilobulated masses were observed, composed of cells arranged in nests similar to aborted follicles or forming cysts, supported by a moderate fibrocollagenous stroma. Pseudorosettes were observed. In mare 2 neoplasm, formations similar to sertoli tubules and cells similar to Leydig cells were observed. In immunohistochemistry, there was inhibin and vimetin marking of mature and neoplastic granulosa cells that made up the nests and lined the cysts. After surgery, mare 1 no longer exhibited behavioral changes, presented heat twice, became pregnant and miscarried. Mare 3 also became pregnant and gave birth to term. There is no information regarding mares 2 after surgery. It was possible to verify with this dissertation that although most neoplasms diagnosed in horses in the Animal Pathology Laboratory of the University Veterinary Hospital of UFCG involve organs related to the integument, neoplasms that affect internal organs occur sporadically and it is important to include them as differential diagnoses.

Goat ovarian follicles express different levels of mRNA for inhibin-ßA subunit and activin-A stimulates secondary follicle growth in vitro

This study investigated the levels of messenger ribonucleic acids (mRNA) for inhibin-ßA subunit in goat primordial, primary and secondary follicles, as well as in cumulus-oocyte complexes (COCs) and mural granulosa / theca cells of antral follicles. The effects of activin-A (100ng mL-1) and/or follicle stimulating hormone (FSH, 50ng mL-1) on growth and expression of mRNA for activin-A and FSH receptor (FSH-R) in secondary follicles cultured for six days were evaluated. The data showed that the expression of inhibin-ßA is lower in secondary follicles than in primary follicles and is higher in large antral follicles than in small antral follicles. After culture, activin-A and/or FSH promoted growth of secondary follicles, while FSH increased the levels of mRNA for inhibin-ßA, and activin-A increased the levels of FSH-R mRNA. In conclusion, mRNA for inhibin-ßA is expressed at different levels in pre-antral and antral follicles and activin-A acts as a stimulator of the FSH-R expression in goat follicles. On its turn, the expression of inhibin-ßA is stimulated by FSH, which together with activin-A promotes secondary follicle growth in-vitro.

Este estudo investigou os níveis de ácidos ribonucleicos (RNAm) para a subunidade ßA da inibina em folículos primordiais, primários e secundários caprinos, bem como em complexos cumulus-oócitos (CCOs) e células da granulosa mural/teca de folículos antrais. Além disso, avaliaram-se os efeitos da ativina-A (100ng mL-1) e/ou hormônio folículo estimulante (FSH, 50ng mL-1) sobre o crescimento e a expressão do RNAm para inibina-ßA e receptores de FSH (FSH-R) em folículos secundários cultivados por seis dias. Os dados mostraram que a expressão de inibina-ßA é menor em folículos secundários do que em folículos primários e é maior em grandes folículos antrais que nos pequenos folículos antrais. Após o cultivo, ativina-A e/ou FSH promoveram o crescimento de folículos secundários. Enquanto o FSH aumentou os níveis de RNAm para inibina-ßA, a ativina-A aumentou os níveis de RNAm para FSH-R. Em conclusão, a inibina-ßA é expressa em diferentes níveis em folículos pré-antrais e antrais e a ativina-A atua como um estimulador da expressão de FSH-R em folículos caprinos. Por sua vez, a expressão de inibina-ßA é estimulada pelo FSH, que, juntamente com ativina, promove o crescimento de folículos secundários in vitro.

Goat ovarian follicles express different levels of mRNA for inhibin-ßA subunit and activin-A stimulates secondary follicle growth in vitro

This study investigated the levels of messenger ribonucleic acids (mRNA) for inhibin-ßA subunit in goat primordial, primary and secondary follicles, as well as in cumulus-oocyte complexes (COCs) and mural granulosa / theca cells of antral follicles. The effects of activin-A (100ng mL-1) and/or follicle stimulating hormone (FSH, 50ng mL-1) on growth and expression of mRNA for activin-A and FSH receptor (FSH-R) in secondary follicles cultured for six days were evaluated. The data showed that the expression of inhibin-ßA is lower in secondary follicles than in primary follicles and is higher in large antral follicles than in small antral follicles. After culture, activin-A and/or FSH promoted growth of secondary follicles, while FSH increased the levels of mRNA for inhibin-ßA, and activin-A increased the levels of FSH-R mRNA. In conclusion, mRNA for inhibin-ßA is expressed at different levels in pre-antral and antral follicles and activin-A acts as a stimulator of the FSH-R expression in goat follicles. On its turn, the expression of inhibin-ßA is stimulated by FSH, which together with activin-A promotes secondary follicle growth in-vitro.

Este estudo investigou os níveis de ácidos ribonucleicos (RNAm) para a subunidade ßA da inibina em folículos primordiais, primários e secundários caprinos, bem como em complexos cumulus-oócitos (CCOs) e células da granulosa mural/teca de folículos antrais. Além disso, avaliaram-se os efeitos da ativina-A (100ng mL-1) e/ou hormônio folículo estimulante (FSH, 50ng mL-1) sobre o crescimento e a expressão do RNAm para inibina-ßA e receptores de FSH (FSH-R) em folículos secundários cultivados por seis dias. Os dados mostraram que a expressão de inibina-ßA é menor em folículos secundários do que em folículos primários e é maior em grandes folículos antrais que nos pequenos folículos antrais. Após o cultivo, ativina-A e/ou FSH promoveram o crescimento de folículos secundários. Enquanto o FSH aumentou os níveis de RNAm para inibina-ßA, a ativina-A aumentou os níveis de RNAm para FSH-R. Em conclusão, a inibina-ßA é expressa em diferentes níveis em folículos pré-antrais e antrais e a ativina-A atua como um estimulador da expressão de FSH-R em folículos caprinos. Por sua vez, a expressão de inibina-ßA é estimulada pelo FSH, que, juntamente com ativina, promove o crescimento de folículos secundários in vitro.

Goat ovarian follicles express different levels of mRNA for inhibin-ßA subunit and activin-A stimulates secondary follicle growth in vitro

This study investigated the levels of messenger ribonucleic acids (mRNA) for inhibin-ßA subunit in goat primordial, primary and secondary follicles, as well as in cumulus-oocyte complexes (COCs) and mural granulosa / theca cells of antral follicles. The effects of activin-A (100ng mL-1) and/or follicle stimulating hormone (FSH, 50ng mL-1) on growth and expression of mRNA for activin-A and FSH receptor (FSH-R) in secondary follicles cultured for six days were evaluated. The data showed that the expression of inhibin-ßA is lower in secondary follicles than in primary follicles and is higher in large antral follicles than in small antral follicles. After culture, activin-A and/or FSH promoted growth of secondary follicles, while FSH increased the levels of mRNA for inhibin-ßA, and activin-A increased the levels of FSH-R mRNA. In conclusion, mRNA for inhibin-ßA is expressed at different levels in pre-antral and antral follicles and activin-A acts as a stimulator of the FSH-R expression in goat follicles. On its turn, the expression of inhibin-ßA is stimulated by FSH, which together with activin-A promotes secondary follicle growth in-vitro.

Este estudo investigou os níveis de ácidos ribonucleicos (RNAm) para a subunidade ßA da inibina em folículos primordiais, primários e secundários caprinos, bem como em complexos cumulus-oócitos (CCOs) e células da granulosa mural/teca de folículos antrais. Além disso, avaliaram-se os efeitos da ativina-A (100ng mL-1) e/ou hormônio folículo estimulante (FSH, 50ng mL-1) sobre o crescimento e a expressão do RNAm para inibina-ßA e receptores de FSH (FSH-R) em folículos secundários cultivados por seis dias. Os dados mostraram que a expressão de inibina-ßA é menor em folículos secundários do que em folículos primários e é maior em grandes folículos antrais que nos pequenos folículos antrais. Após o cultivo, ativina-A e/ou FSH promoveram o crescimento de folículos secundários. Enquanto o FSH aumentou os níveis de RNAm para inibina-ßA, a ativina-A aumentou os níveis de RNAm para FSH-R. Em conclusão, a inibina-ßA é expressa em diferentes níveis em folículos pré-antrais e antrais e a ativina-A atua como um estimulador da expressão de FSH-R em folículos caprinos. Por sua vez, a expressão de inibina-ßA é estimulada pelo FSH, que, juntamente com ativina, promove o crescimento de folículos secundários in vitro.

Folliculogenesis in ewes: estimulation and inhibition factors of follicular growth / Foliculogênese em ovelhas: fatores de estímulo e inibição do crescimento folicular

The regulatory mechanisms of females´ reproduction are crucial to the success of conventional or assisted breeding. The follicle is the functional unit of the ovary of the female mammal species, being its major functions the growth and maturation of oocytes, as well as the production of steroid hormones. Several factors act jointly on the regulation of these activities through endocrine, autocrine and pa­racrine mechanisms stimulating the mitogenic activity of quiescent oocytes and also steroidogenic activity of the theca cells and of the granulosa. Among the regulatory factors, there are some that act in a stimulant way, such as the receptor of the binding protein (c-KITL), the system of growth factors related to insulin (IGFs), the activin and follistatin complex, and the super family of growth- transforming factor (TGF-). Other agents are inhibitory, such as inhibin, the retinoblastoma protein, expressed in oocytes, as the Wilms gene (WTI). This paper aims to review the regulatory mecha¬nisms of folliculogenesis and its importance in the reproductive physiology of ewes.

Os mecanismos de regulação da reprodução de fêmeas são de impor­tância fundamental para o sucesso da reprodução convencional ou assistida. O folículo é a unidade funcional do ovário de fêmeas das espécies mamíferas, tendo como principais funções o crescimento e a maturação dos oócitos, bem como a produção de hormônios esteróides. Vários fatores agem de forma conjunta na regulação destas atividades por meio de mecanismos endócrinos, autócrinos e parácrinos, estimulando a atividade mitogênica de oócitos quiescentes e, ativi­dade esteroidogênica de células da teca e da granulosa. Entre os fatores regu­latórios existem alguns que agem de forma estimulante, tais como o receptor da proteína ligante (c-KITL), o sistema de fatores de crescimento ligados à insulina (IGFs), o complexo ativina e folistatina e a superfamília do fator transformador de crescimento- (TGF-). Outros fatores agem de forma inibitória, tais como a inibi­na, a proteína retinoblastoma, expressa em oócitos, igualmente ao gene de Wilms (WTI). No presente trabalho, objetivou-se abordar os mecanismos regulatórios da foliculogênese e sua importância na fisiologia reprodutiva de ovelhas.

Ativina intraovariana: uma atualização sobre estrutura, regulação e função / Intraovarian activin: an update on structure, regulation and function

Background: Important advances have been made recently that clarify our understanding of the structural basis, signaling and regulation, as well as the biological role of activin in ovaries. During folliculogenesis various growth factors are produced locally in the mammalian ovary. Among these factors, activin has been a focal point in research as it has emerged as a crucial substance capable of inducing follicular development. The important actions indicate that activin has many relevant homeostatic functions in the reproduction of several species. Therefore, this review discusses the ligand protein structure, activin receptors, mechanisms of action and regulation, as well as the importance of activin on in vitro culture of preantral follicles. Review: Activin belongs to the transforming growth factor β (TGF - β) super family. It is a homodimer or heterodimer of two similar but distinct subunits (βA and βB). The dimerisation of activin subunits gives rise to three forms of activin, which are classified as, activin A (βA - βA), activin B (βB - βB) and activin AB (βA - βB). The biological activity of activin occurs through its connection with two types of cell surface receptors designated type I and type II. These receptors are represented by two isoforms, activin receptor types IA (ActR - IA), IB (ActR - IB), IIA (ActR - IIA) and IIB (ActR - IIB). Activin receptors are transmembrane proteins, composed of a ligand-binding extracellular domain, a transmembrane domain and a cytoplasmic domain with serine/threonine kinase activity. The transient activation of the receptor induces phosphorylation of protein mediators called Smads. Activation of Smad 2/3 by phosphorylation causes trimerization and hetero-oligomerization with the common Smad, Smad 4. This complex translocates to the nucleus to activating and regulating transcription of target genes. Members of another class of Smads act as negative regulators of the signal transduction pathway. Inhibitory Smad 7 can bind to type I receptors, preventing receptor–Smad 2/3 association, or by competitively binding of Smad 4, which blocks Smad intracellular translocation. In addition, within the extracellular environment, binding proteins such as follistatin and inhibin can modulate the biological activity of activin. In the ovaries of mammals specifically, activin participates in several cellular events, including cellular proliferation, differentiation, and survival, as well as assisting steroidal hormones during follicular development. Activin has been localized in the oocytes and granulosa cells of rodent, porcine, caprine and bovine follicles. Activin is also within the granulosa cells of human follicles and in the thecal layers of porcine and human. In addition, activin stimulates follicle growth in-vitro, is used in pre-antral ovine and caprine follicles and enhances growth and survival of human pre-antral follicles in vitro. Conclusion: Activin is controlled by competitive substances and a dynamic interaction between the various regulatory proteins responsible for coordinating several signaling pathways. The balance between the actions of these proteins is critical for regulation of gene expression in different structures, including pre-antral follicles. However, the nature of physiological effects of activin in the ovary is still equivocal and awaits clarification.(AU)

Ativina intraovariana: uma atualização sobre estrutura, regulação e função / Intraovarian activin: an update on structure, regulation and function

Background: Important advances have been made recently that clarify our understanding of the structural basis, signaling and regulation, as well as the biological role of activin in ovaries. During folliculogenesis various growth factors are produced locally in the mammalian ovary. Among these factors, activin has been a focal point in research as it has emerged as a crucial substance capable of inducing follicular development. The important actions indicate that activin has many relevant homeostatic functions in the reproduction of several species. Therefore, this review discusses the ligand protein structure, activin receptors, mechanisms of action and regulation, as well as the importance of activin on in vitro culture of preantral follicles. Review: Activin belongs to the transforming growth factor β (TGF - β) super family. It is a homodimer or heterodimer of two similar but distinct subunits (βA and βB). The dimerisation of activin subunits gives rise to three forms of activin, which are classified as, activin A (βA - βA), activin B (βB - βB) and activin AB (βA - βB). The biological activity of activin occurs through its connection with two types of cell surface receptors designated type I and type II. These receptors are represented by two isoforms, activin receptor types IA (ActR - IA), IB (ActR - IB), IIA (ActR - IIA) and IIB (ActR - IIB). Activin receptors are transmembrane proteins, composed of a ligand-binding extracellular domain, a transmembrane domain and a cytoplasmic domain with serine/threonine kinase activity. The transient activation of the receptor induces phosphorylation of protein mediators called Smads. Activation of Smad 2/3 by phosphorylation causes trimerization and hetero-oligomerization with the common Smad, Smad 4. This complex translocates to the nucleus to activating and regulating transcription of target genes. Members of another class of Smads act as negative regulators of the signal transduction pathway. Inhibitory Smad 7 can bind to type I receptors, preventing receptor–Smad 2/3 association, or by competitively binding of Smad 4, which blocks Smad intracellular translocation. In addition, within the extracellular environment, binding proteins such as follistatin and inhibin can modulate the biological activity of activin. In the ovaries of mammals specifically, activin participates in several cellular events, including cellular proliferation, differentiation, and survival, as well as assisting steroidal hormones during follicular development. Activin has been localized in the oocytes and granulosa cells of rodent, porcine, caprine and bovine follicles. Activin is also within the granulosa cells of human follicles and in the thecal layers of porcine and human. In addition, activin stimulates follicle growth in-vitro, is used in pre-antral ovine and caprine follicles and enhances growth and survival of human pre-antral follicles in vitro. Conclusion: Activin is controlled by competitive substances and a dynamic interaction between the various regulatory proteins responsible for coordinating several signaling pathways. The balance between the actions of these proteins is critical for regulation of gene expression in different structures, including pre-antral follicles. However, the nature of physiological effects of activin in the ovary is still equivocal and awaits clarification.

Endocrine dynamics associated with follicle development in pigs: a review

A significant increase in growth of follicles that are selected for ovulation seems to occur on days 14-17 of the estrous cycle. In pigs there is a continuous growth of follicles without appearance of dominant follicles or follicle waves during the estrous cycle. There is a general consensus that a decrease of FSH during the follicular phase is accompanied with the selection of ovulatory follicles and changing from FSH to LH dependence. Development of preovulatory follicles is prevented during lactation mainly due to the inhibition of LH secretion. FSH is not inhibited during lactation and temporary increase in FSH is associated with wave-like follicular growth. Weaning of piglets normally results in increased secretion of LH, which is characterized by a high pulse frequency and low pulse amplitude. The duration of weaning-to-estrus interval is associated with plasma changes in gonadotropins, steroids, inhibin, leptin, IGF-I and insulin. Evidence for a positive role of short elevations in cortisol on LH secretion and ovarian function are accumulating but further studies are still needed to elucidate this issue. The aim of this review was to summarize our current knowledge of endocrinological changes in relation to follicular development during estrous cycle, lactation and after weaning in pigs.

Endocrine dynamics associated with follicle development in pigs: a review

A significant increase in growth of follicles that are selected for ovulation seems to occur on days 14-17 of the estrous cycle. In pigs there is a continuous growth of follicles without appearance of dominant follicles or follicle waves during the estrous cycle. There is a general consensus that a decrease of FSH during the follicular phase is accompanied with the selection of ovulatory follicles and changing from FSH to LH dependence. Development of preovulatory follicles is prevented during lactation mainly due to the inhibition of LH secretion. FSH is not inhibited during lactation and temporary increase in FSH is associated with wave-like follicular growth. Weaning of piglets normally results in increased secretion of LH, which is characterized by a high pulse frequency and low pulse amplitude. The duration of weaning-to-estrus interval is associated with plasma changes in gonadotropins, steroids, inhibin, leptin, IGF-I and insulin. Evidence for a positive role of short elevations in cortisol on LH secretion and ovarian function are accumulating but further studies are still needed to elucidate this issue. The aim of this review was to summarize our current knowledge of endocrinological changes in relation to follicular development during estrous cycle, lactation and after weaning in pigs.(AU)

Human antral folliculogenesis: what we have learned from the bovine and equine models

The study of ovarian folliculogenesis has been of great interest to scientists and clinicians in the human and veterinary health fields for more than 20 centuries. Initial studies of the ovarian follicle were based on anatomical descriptions post-mortem, followed by histologic and endocrinologic evaluation of ovarian status. The introduction of high resolution ultrasonography in the 1980s provided a long-awaited tool to image the reproductive tissues in situ in both animal and human species. The bovine and equine species have been established as models for the study of human ovarian folliculogenesis. Profound similarities in the dynamics of follicle development exist between the menstrual cycle in humans and the estrous cycle in cattle and horses. Disparities between species appear specific rather than general. Research performed in women thus far has led to the concepts that: 1) follicle development occurs in a wave-like manner during the menstrual cycle, 2) the number of waves per cycle correlates positively with the length of the cycle, 3) the emergence of follicle waves in women are preceded by a rise in circulating FSH, 4) selection of a dominant follicle may occur in each wave of the cycle, and 5) a decline in circulating FSH and increase in follicular estradiol, inhibin A, and IGF-II act collectively to enable the dominant follicle to continue to grow in an endocrine environment of decreasing FSH and increasing LH, while subordinate follicles undergo regression. The goal of continued research using animal models for studying human ovarian function is to provide the hypothetical basis for further studies in women, which will ultimately lead to the development of safer and more efficacious infertility and contraceptive therapies.

Human antral folliculogenesis: what we have learned from the bovine and equine models

The study of ovarian folliculogenesis has been of great interest to scientists and clinicians in the human and veterinary health fields for more than 20 centuries. Initial studies of the ovarian follicle were based on anatomical descriptions post-mortem, followed by histologic and endocrinologic evaluation of ovarian status. The introduction of high resolution ultrasonography in the 1980s provided a long-awaited tool to image the reproductive tissues in situ in both animal and human species. The bovine and equine species have been established as models for the study of human ovarian folliculogenesis. Profound similarities in the dynamics of follicle development exist between the menstrual cycle in humans and the estrous cycle in cattle and horses. Disparities between species appear specific rather than general. Research performed in women thus far has led to the concepts that: 1) follicle development occurs in a wave-like manner during the menstrual cycle, 2) the number of waves per cycle correlates positively with the length of the cycle, 3) the emergence of follicle waves in women are preceded by a rise in circulating FSH, 4) selection of a dominant follicle may occur in each wave of the cycle, and 5) a decline in circulating FSH and increase in follicular estradiol, inhibin A, and IGF-II act collectively to enable the dominant follicle to continue to grow in an endocrine environment of decreasing FSH and increasing LH, while subordinate follicles undergo regression. The goal of continued research using animal models for studying human ovarian function is to provide the hypothetical basis for further studies in women, which will ultimately lead to the development of safer and more efficacious infertility and contraceptive therapies.(AU)

Concentrações hormonais e desenvolvimento folicular de vacas leiteiras em hipertermia sazonal e aguda / Hormone concentration and follicular development in dairy cows under seasonal and acute hyperthermia

Avaliaram-se as concentrações hormonais e os parâmetros de desenvolvimento folicular de vacas leiteiras expostas ao calor sazonal e agudo. Dividiram-se os animais em quatro grupos: verão (n=5), outono (n=5), inverno com hipertermia aguda (grupo câmara climática, (CC), n=5) e inverno (n=9). Os animais foram abatidos no sétimo dia após a ovulação, e os parâmetros de desenvolvimento folicular avaliados. O líquido folicular do maior folículo foi aspirado e armazenado para posterior análise de hormônios esteróides e inibina. O número de células da granulosa vivas no verão e no outono foi 40 e 45% respectivamente, menor que no inverno (P<0,05). A concentração de estradiol (E2) no inverno foi 62% maior que no outono (P<0,05) e 34% superior ao grupo verão (P<0,06). Houve um aumento na quantidade de androstenediona no verão em relação aos grupos inverno (P<0,08) e outono (P<0,05). A concentração de inibina foi maior no inverno do que no verão e CC (P<0,05). A exposição ao calor sazonal e agudo modificou os parâmetros de desenvolvimento do folículo e as concentrações hormonais no líquido folicular, podendo explicar em parte a queda nas taxas de concepção no verão.(AU)

The present study evaluated the seasonal and acute heat stress on follicular development and steroid and inhibin concentrations in follicular fluid, in bovine dominant follicle. Cows were distributed into four treatments: summer (n=5), autumn (n=5), animals heat stressed during the winter (n=5) and winter (n=9). On day 7 of the estrous cycle, animals were slaughtered and parameters related to follicle development were evaluated. The follicular fluid (FF) was aspirated and stored for further hormonal analysis. During the summer, the number of viable granulosa cells was 40% lower than during the winter, and there was a 45% decrease in this parameter during the autumn (P<0.05). In the winter, estradiol concentration was 62% higher than during the autumn (P<0.05) and 42% higher than during the summer (P<0.06). There was an increase in androstenedione concentration in summer group, when compared to winter (P<0.08) and autumn (P<0.05) groups. Inhibin concentration was higher in winter groups than summer and winter heat stressed groups (P<0.05). Seasonal and acute heat stress altered developmental parameters in dominant follicle and hormonal concentration in follicular fluid, those effects can partially explain the decrease in conception rates during summer.(AU)