Post-partum reproductive activity and estrus synchronization responsiveness in anglonubian x sprd fed with dried carnauba wax palm fruit (Copernicia prunifera) long term / Atividade reprodutiva e resposta à sincronização de estro no pós-parto em cabras mestiças de anglonubiana x sprd alimentadas com o fruto desidratado da carnaubeira (Copercinia prunifera) por longo tempo

The present study evaluates the use of dried carnauba wax palm fruit in 28 postpartum goats fed diets containing Bermudagrass hay and either corn (WCWP) or carnauba-based (CWP) concentrated feed. Estrus synchronization was performed 90 days postpartum, and the goats were mated. In the CWP group, compared to the WCWP group, the daily dry matter intake was significantly reduced (646.48 ± 11.03 g vs. 739.29 ± 3.24 g, respectively; P 0.01). The time to the first functional corpus luteum was similar between the groups, occurring a mean 66 days postpartum. During estrus synchronization, the CWP group had a decreased sternal subcutaneous adipose tissue thickness when compared to the WCWP group (11.93 ± 0.45 mm vs. 13.69 ± 0.57 mm, respectively; P 0.05) and a lower pregnancy rate (86.67% vs. 46.15%, respectively; P 0.02). The embryonic vesicle and crown-rump length growth rates, and the biparietal, thoracic, and abdominal diameters at 45 days of pregnancy were within normal range for goats in both groups. Litter size at birth was similar between the groups at a mean 1.39 ± 0.11. In conclusion, the substitution of corn with dehydrated carnauba wax palm fruit in concentrated feed for postpartum goats showed no positive effects. Reduced feed intake in the does consuming the carnauba diet caused decreased body reserves, which probably resulted in a decreased pregnancy rate...(AU)

O presente estudo avaliou o uso do fruto da carnaubeira desidratado em 28 cabras no período pós-parto alimentadas com dieta contendo feno de Tifton e concentrado comercial a base de milho (WCWP) ou com o fruto da carnaubeira (CWP). A sincronização de estro foi realizada aos 90 dias pós-parto e todas as fêmeas foram cobertas. No grupo CWP, em relação ao grupo WCWP, o consumo de matéria seca foi significativamente menor (646.48 ± 11.03 g vs. 739.29 ± 3.24 g, respectivamente; P 0.01). O tempo para o aparecimento do primeiro corpo lúteo funcional foi semelhante entre os grupos, ocorrendo em média aos 66 dias pós-parto. No momento da sincronização do estro, o grupo CWP apresentou uma menor espessura do tecido adiposo subcutâneo esternal quando comparado ao grupo WCWP (13,69 ± 0,57 mm vs. 11,93 ± 0,45 mm, respectivamente; P 0,05) e uma menor taxa de gestação (86,67% vs. 46,15%, respectivamente; P 0.02). A taxa de crescimento da vesícula embrionária e do comprimento crânio-caudal, assim como no diâmetro biparietal, torácico e abdominal aos 45 dias de gestação estavam em ambos os grupos dentro do esperado para a espécie em questão. Ao parto a prolificidade não diferiu entre os grupos, com média de 1,39 ± 0,11. Em conclusão, a substituição total do milho pelo fruto da carnaubeira no concentrado fornecido às fêmeas caprinas no período pós-parto não apresentou efeitos...(AU)

Caracterização estrutural do fator de crescimento de fibroblasto-10 (FGF-10) e seu papel na fisiologia folicular ovariana / Structural characterization of the fibroblast growth factor-10 (FGF-10) and its role in ovarian follicular physiology

Background: Interest in folliculogenesis has grown extensively in recent years. Nevertheless, several aspects of follicular activity are still poorly understood. Thus, in vitro culture of ovarian follicles using new substances has been established as a very viable model, enhancing the prospects for a better understanding of follicular activity. Among the family members of the fibroblast growth factor (FGFs), FGF-10 has received recent attention for its ability to regulate the development of ovarian follicles and oocyte maturation. Given the relevance of FGF-10 in the folliculogenesis process, this review aimed to describe the structural features, expression and the main biological effects of FGF-10 on the development of ovarian follicles in mammals. Review: Along this work, it was shown aspects related to structural characterization of FGF-10 and its receptors, as well as FGF-10 expression in different cell types, emphasizing its importance to follicular development. FGF-10 is a paracrine member of the family of FGFs, and is characterized by promoting biological responses via cell surface receptors (FGFRs) of tyrosine kinase-type. Of these receptors, FGFR-1, FGFR-2 and FGFR-3 may undergo alternative transcriptional arrangements, enabling the formation of two isoforms (b and c) that have varying degrees of affinity for the various FGFs. Thus, seven FGFR proteins (FGFRs 1b, 1c, 2b, 2c, 3b, 3c and 4) with different binding specificities are generated from the four FGFR genes. The FGFRs transmit intracellular signals after binding with the ligand through the phosphorylation of tyrosine, which activates various transduction patterns in the cytoplasm. The signal transduction of FGF-10 may occur through three main pathways: protein of rat sarcoma (Ras)/MAPK, PLCg/Ca2+ and phosphatidylinositol-3 kinase (PI3K)/ protein kinase B (Akt), which are involved in the transmission of biological signals, leading to cellular proliferation and differentiation. FGF-10 mRNA expression was detected in the ovarian stroma, oocyte and theca cells of preantral and antral follicles. On the other hand, the expression of mRNA for FGF-10 receptors was found in, granulosa cells, theca cells, cumulus cells and oocytes. Although FGFs are widely distributed in different tissues and cell types, the importance and function of FGFs in the ovary are still poorly documented. FGF-10 has been shown to be an important mediator of mesenchymal and epithelial cell interactions during follicle development, promoting follicular survival, activation and growth. Besides the action on folliculogenesis, FGF-10 was recently identified as a growth factor able to improve oocyte competence. However, in antral follicles, the presence of FGF-10 is associated with increased follicular atresia, which matches its anti-estrogenic action. Discussion: From this review, we can conclude that FGF-10 is an important regulator of female reproduction. This growth factor acts in follicle survival, oocyte maturation, expansion of cumulus cells and proliferation of granulosa/theca cells through direct and/or indirect actions in the control of folliculogenesis. Furthermore, FGF-10 seemed to have different effects throughout the follicular development. However, it is necessary to perform additional studies that may provide a better understanding about the importance of FGF-10 during folicullogenesis.(AU)

Caracterização estrutural do fator de crescimento de fibroblasto-10 (FGF-10) e seu papel na fisiologia folicular ovariana / Structural characterization of the fibroblast growth factor-10 (FGF-10) and its role in ovarian follicular physiology

Background: Interest in folliculogenesis has grown extensively in recent years. Nevertheless, several aspects of follicular activity are still poorly understood. Thus, in vitro culture of ovarian follicles using new substances has been established as a very viable model, enhancing the prospects for a better understanding of follicular activity. Among the family members of the fibroblast growth factor (FGFs), FGF-10 has received recent attention for its ability to regulate the development of ovarian follicles and oocyte maturation. Given the relevance of FGF-10 in the folliculogenesis process, this review aimed to describe the structural features, expression and the main biological effects of FGF-10 on the development of ovarian follicles in mammals. Review: Along this work, it was shown aspects related to structural characterization of FGF-10 and its receptors, as well as FGF-10 expression in different cell types, emphasizing its importance to follicular development. FGF-10 is a paracrine member of the family of FGFs, and is characterized by promoting biological responses via cell surface receptors (FGFRs) of tyrosine kinase-type. Of these receptors, FGFR-1, FGFR-2 and FGFR-3 may undergo alternative transcriptional arrangements, enabling the formation of two isoforms (b and c) that have varying degrees of affinity for the various FGFs. Thus, seven FGFR proteins (FGFRs 1b, 1c, 2b, 2c, 3b, 3c and 4) with different binding specificities are generated from the four FGFR genes. The FGFRs transmit intracellular signals after binding with the ligand through the phosphorylation of tyrosine, which activates various transduction patterns in the cytoplasm. The signal transduction of FGF-10 may occur through three main pathways: protein of rat sarcoma (Ras)/MAPK, PLCg/Ca2+ and phosphatidylinositol-3 kinase (PI3K)/ protein kinase B (Akt), which are involved in the transmission of biological signals, leading to cellular proliferation and differentiation. FGF-10 mRNA expression was detected in the ovarian stroma, oocyte and theca cells of preantral and antral follicles. On the other hand, the expression of mRNA for FGF-10 receptors was found in, granulosa cells, theca cells, cumulus cells and oocytes. Although FGFs are widely distributed in different tissues and cell types, the importance and function of FGFs in the ovary are still poorly documented. FGF-10 has been shown to be an important mediator of mesenchymal and epithelial cell interactions during follicle development, promoting follicular survival, activation and growth. Besides the action on folliculogenesis, FGF-10 was recently identified as a growth factor able to improve oocyte competence. However, in antral follicles, the presence of FGF-10 is associated with increased follicular atresia, which matches its anti-estrogenic action. Discussion: From this review, we can conclude that FGF-10 is an important regulator of female reproduction. This growth factor acts in follicle survival, oocyte maturation, expansion of cumulus cells and proliferation of granulosa/theca cells through direct and/or indirect actions in the control of folliculogenesis. Furthermore, FGF-10 seemed to have different effects throughout the follicular development. However, it is necessary to perform additional studies that may provide a better understanding about the importance of FGF-10 during folicullogenesis.

Avanços no isolamento e sistemas de cultivo de folículos pré-antrais / Advances in isolation and culture systems of preantral follicles

Os folículos pré-antrais presentes no córtex ovariano representam cerca de 95% de toda a população folicular e podem ser crescidos e maturados in vitro, visando a obtenção posterior de oócitos maturos e competentes, os quais poderão ser utilizados nas diversas biotécnicas da reprodução aplicadas aos animais domésticos. Os estudos relacionados com a foliculogênese podem ser feitos in vivo, através da análise de expressão de genes ou proteínas que estão presentes no ovário, e ainda in vitro com a utilização de diferentes sistemas, onde os folículos podem ser cultivados in situ ou isolados. Deste modo, neste trabalho foram abordados os diferentes métodos de isolamento e sistemas de cultivo in vitro empregados para a manutenção da integridade estrutural e desenvolvimento de folículos pré-antrais.

The preantral follicles enclosed in ovarian cortex represent about 95% of the entire follicular population and can be grown and matured in vitro in order to obtain mature and competent oocytes, which may be used in various reproductive biotechnologies applied to domestic animals. Studies related to folliculogenesis can be performed in vivo, by analyzing the expression of genes or proteins that are present in the ovary, and also in vitro with the use of different systems, in which the follicles can be grown in situ or isolated. Thus, this study shows the different methods of isolation and in vitro systems employed for maintaining the structural integrity and development of preantral follicles.

Caracterização estrutural e papel do fator de crescimento derivado de plaquetas (PDGF) na foliculogênese ovariana / Structural Characterization and Role of Platelet-Derived Growth Factor (PDGF) in Ovarian Folliculogenesis

Background: The platelet-derived growth factor (PDGF) is expressed in a wide variety of cell types, exerts a potent mitogenic role and acts on the growth, differentiation and cell chemotaxis. Studies have shown that during folliculogenesis, PDGF and their receptors are expressed in oocytes, granulosa cells and thecal cells of ovarian follicles at different developmental stages in several species. Although exist many information about its expression sites, as well as about its action in different cells types, the role of PDGF on ovarian folliculogenesis remains understudied. Thus, this article aims to review issues related to PDGF, suggesting the involvement of this mitogenic factor during follicular development. Review: Along this work, it was shown aspects related to structural characterization of PDGF and its receptors, as well as PDGF expression in different cells types, emphasizing its importance to follicular development. PDGF family is composed by four polypeptide chains (each encoded by a different gene), which are synthesized in the form of inactive pro-proteins. After a proteolytic processing, these chains undergo homo or heterodimerization, resulting in five isoforms (PDGF-AA, -BB, -AB, -CC e -DD). The cellular effects of these different PDGF isoforms are mediated by binding, with different specificities, to three transmembrane receptors isoforms of type tyrosine kinase generated by the association of subunits alpha e beta (PDGFR- alphaalpha, - alphabeta, - betabeta)...

Caracterização estrutural e papel do fator de crescimento derivado de plaquetas (PDGF) na foliculogênese ovariana / Structural Characterization and Role of Platelet-Derived Growth Factor (PDGF) in Ovarian Folliculogenesis

Background: The platelet-derived growth factor (PDGF) is expressed in a wide variety of cell types, exerts a potent mitogenic role and acts on the growth, differentiation and cell chemotaxis. Studies have shown that during folliculogenesis, PDGF and their receptors are expressed in oocytes, granulosa cells and thecal cells of ovarian follicles at different developmental stages in several species. Although exist many information about its expression sites, as well as about its action in different cells types, the role of PDGF on ovarian folliculogenesis remains understudied. Thus, this article aims to review issues related to PDGF, suggesting the involvement of this mitogenic factor during follicular development. Review: Along this work, it was shown aspects related to structural characterization of PDGF and its receptors, as well as PDGF expression in different cells types, emphasizing its importance to follicular development. PDGF family is composed by four polypeptide chains (each encoded by a different gene), which are synthesized in the form of inactive pro-proteins. After a proteolytic processing, these chains undergo homo or heterodimerization, resulting in five isoforms (PDGF-AA, -BB, -AB, -CC e -DD). The cellular effects of these different PDGF isoforms are mediated by binding, with different specificities, to three transmembrane receptors isoforms of type tyrosine kinase generated by the association of subunits alpha e beta (PDGFR- alphaalpha, - alphabeta, - betabeta)...(AU)

Desenvolvimento in vitro de folículos pré-antrais caprinos na presença de kit ligand (kl) e proteína morfogenética óssea 15 (bmp-15) / Development of in vitro preantral follicles goats in the presence of kit ligand (kl) and protein bone morphogenetic 15 (bmp-15)

Esse estudo teve como objetivos: 1) quantificar os níveis de RNAm para os receptores de kit ligand (KL) e proteína morfogenética óssea 15 (BMP-15) em folículos ovarianos caprinos utilizando RT-PCR em tempo real; 2) investigar o papel do KL, associado ou não ao hormônio folículo estimulante (FSH) em meio dinâmico, sobre a sobrevivência, ativação e crescimento de folículos pré-antrais caprinos cultivados in situ; 3) avaliar os efeitos do KL e da BMP-15 sobre a sobrevivência, formação de antro e crescimento in vitro de folículos secundários caprinos isolados, bem como sobre a retomada da meiose de seus oócitos. [...] Verificou-se ainda níveis maiores de RNAm para o receptor tipo I da BMP-15 (BMPRIB) nas células da granulosa/teca de grandes folículos antrais em comparação àquelas de pequenos antrais. Após 16 dias de cultivo in situ, observou-se que o meio dinâmico contendo KL até o dia 8 e FSH do dia 8 ao dia 16 manteve a integridade ultraestrutural e induziu a ativação e o crescimento folicular. Quanto aos folículos isolados, a adição de 50 e/ou 100 ng/mL de KL ao cultivo, na ausência de FSH, promoveu a sobrevivência e melhorou a formação de antro, crescimento oocitário e retomada da meiose após 18 dias. Já 50 ng/mL de BMP-15 manteve a ultraestrutura e estimulou a formação de antro e o crescimento de folículos isolados. Em conclusão, os resultados deste estudo mostraram que os RNAm para c-kit, BMPRIB e BMPRII foram detectados em todas as categorias foliculares e tipos celulares investigados. A utilização de KL e FSH de forma dinâmica em cultivo in situ, bem como de KL (50 e/ou 100 ng/mL) e BMP-15 (50 ng/mL) em cultivo de folículos secundários isolados, promoveu a manutenção da sobrevivência folicular e o desenvolvimento de folículos pré-antrais caprinos.

The aims of this study were: 1) to quantify the mRNA levels for kit ligand (KL) and bone morphogenetic protein 15 (BMP-15) receptors in goat ovarian follicles by real time RT-PCR; 2) to investigate the role of KL alone or associated with follicle stimulating hormone (FSH) in a dynamic medium on the survival, activation and growth of goat preantral follicles cultured in situ; 3) to evaluate the effects of KL and BMP-15 on the in vitro survival, antrum formation and growth of goat isolated secondary follicles, and on the meiotic resumption of their oocytes. [...] Moreover, higher mRNA levels were observed for ckit in cumulus-oocyte complexes from small and large antral follicles compared with their respective granulosa/theca cells, which also occurred for BMPRII in the large antral follicles. Higher mRNA levels were also seen for BMP-15 type I receptor (BMPRIB) in the granulosa/theca cells from large antral follicles compared with those from small antral follicles. After 16 days of in situ culture, the dynamic medium containing KL until day 8 and FSH from day 8 to day 16 maintained the ultrastructural integrity and induced activation and follicular growth. For the isolated follicles, the addition of 50 and/or 100 ng/mL of KL to the culture, in the absence of FSH, promoted follicle survival and improved antrum formation,oocyte growth and meiotic resumption after 18 days. BMP-15 (50 ng/mL) maintained the ultrastructure and stimulated antrum formation and growth in isolated follicles. In conclusion, the results of this study showed that mRNAs for c-kit, BMPRIB and BMPRII were detected in all follicular categories and cellular types investigated. The use of KL and FSH dynamically in in situ culture, and the use of KL (50 and/or 100 ng/mL) and BMP-15 (50 ng/mL) in culture of isolated secondary follicles, promoted the maintenance of follicular survival and the development of caprine preantral follicles.

Desenvolvimento in vitro de folículos pré-antrais caprinos na presença de kit ligand (kl) e proteína morfogenética óssea 15 (bmp-15) / Development of in vitro preantral follicles goats in the presence of kit ligand (kl) and protein bone morphogenetic 15 (bmp-15)

Esse estudo teve como objetivos: 1) quantificar os níveis de RNAm para os receptores de kit ligand (KL) e proteína morfogenética óssea 15 (BMP-15) em folículos ovarianos caprinos utilizando RT-PCR em tempo real; 2) investigar o papel do KL, associado ou não ao hormônio folículo estimulante (FSH) em meio dinâmico, sobre a sobrevivência, ativação e crescimento de folículos pré-antrais caprinos cultivados in situ; 3) avaliar os efeitos do KL e da BMP-15 sobre a sobrevivência, formação de antro e crescimento in vitro de folículos secundários caprinos isolados, bem como sobre a retomada da meiose de seus oócitos. [...] Verificou-se ainda níveis maiores de RNAm para o receptor tipo I da BMP-15 (BMPRIB) nas células da granulosa/teca de grandes folículos antrais em comparação àquelas de pequenos antrais. Após 16 dias de cultivo in situ, observou-se que o meio dinâmico contendo KL até o dia 8 e FSH do dia 8 ao dia 16 manteve a integridade ultraestrutural e induziu a ativação e o crescimento folicular. Quanto aos folículos isolados, a adição de 50 e/ou 100 ng/mL de KL ao cultivo, na ausência de FSH, promoveu a sobrevivência e melhorou a formação de antro, crescimento oocitário e retomada da meiose após 18 dias. Já 50 ng/mL de BMP-15 manteve a ultraestrutura e estimulou a formação de antro e o crescimento de folículos isolados. Em conclusão, os resultados deste estudo mostraram que os RNAm para c-kit, BMPRIB e BMPRII foram detectados em todas as categorias foliculares e tipos celulares investigados. A utilização de KL e FSH de forma dinâmica em cultivo in situ, bem como de KL (50 e/ou 100 ng/mL) e BMP-15 (50 ng/mL) em cultivo de folículos secundários isolados, promoveu a manutenção da sobrevivência folicular e o desenvolvimento de folículos pré-antrais caprinos.(AU)

The aims of this study were: 1) to quantify the mRNA levels for kit ligand (KL) and bone morphogenetic protein 15 (BMP-15) receptors in goat ovarian follicles by real time RT-PCR; 2) to investigate the role of KL alone or associated with follicle stimulating hormone (FSH) in a dynamic medium on the survival, activation and growth of goat preantral follicles cultured in situ; 3) to evaluate the effects of KL and BMP-15 on the in vitro survival, antrum formation and growth of goat isolated secondary follicles, and on the meiotic resumption of their oocytes. [...] Moreover, higher mRNA levels were observed for ckit in cumulus-oocyte complexes from small and large antral follicles compared with their respective granulosa/theca cells, which also occurred for BMPRII in the large antral follicles. Higher mRNA levels were also seen for BMP-15 type I receptor (BMPRIB) in the granulosa/theca cells from large antral follicles compared with those from small antral follicles. After 16 days of in situ culture, the dynamic medium containing KL until day 8 and FSH from day 8 to day 16 maintained the ultrastructural integrity and induced activation and follicular growth. For the isolated follicles, the addition of 50 and/or 100 ng/mL of KL to the culture, in the absence of FSH, promoted follicle survival and improved antrum formation,oocyte growth and meiotic resumption after 18 days. BMP-15 (50 ng/mL) maintained the ultrastructure and stimulated antrum formation and growth in isolated follicles. In conclusion, the results of this study showed that mRNAs for c-kit, BMPRIB and BMPRII were detected in all follicular categories and cellular types investigated. The use of KL and FSH dynamically in in situ culture, and the use of KL (50 and/or 100 ng/mL) and BMP-15 (50 ng/mL) in culture of isolated secondary follicles, promoted the maintenance of follicular survival and the development of caprine preantral follicles.(AU)

Post-partum reproductive activity and estrus synchronization responsiveness in anglonubian x sprd fed with dried carnauba wax palm fruit (Copernicia prunifera) long term / Atividade reprodutiva e resposta à sincronização de estro no pós-parto em cabras mestiças de anglonubiana x sprd alimentadas com o fruto desidratado da carnaubeira (Copercinia prunifera) por longo tempo

The present study evaluates the use of dried carnauba wax palm fruit in 28 postpartum goats fed diets containing Bermudagrass hay and either corn (WCWP) or carnauba-based (CWP) concentrated feed. Estrus synchronization was performed 90 days postpartum, and the goats were mated. In the CWP group, compared to the WCWP group, the daily dry matter intake was significantly reduced (646.48 ± 11.03 g vs. 739.29 ± 3.24 g, respectively; P 0.01). The time to the first functional corpus luteum was similar between the groups, occurring a mean 66 days postpartum. During estrus synchronization, the CWP group had a decreased sternal subcutaneous adipose tissue thickness when compared to the WCWP group (11.93 ± 0.45 mm vs. 13.69 ± 0.57 mm, respectively; P 0.05) and a lower pregnancy rate (86.67% vs. 46.15%, respectively; P 0.02). The embryonic vesicle and crown-rump length growth rates, and the biparietal, thoracic, and abdominal diameters at 45 days of pregnancy were within normal range for goats in both groups. Litter size at birth was similar between the groups at a mean 1.39 ± 0.11. In conclusion, the substitution of corn with dehydrated carnauba wax palm fruit in concentrated feed for postpartum goats showed no positive effects. Reduced feed intake in the does consuming the carnauba diet caused decreased body reserves, which probably resulted in a decreased pregnancy rate...

O presente estudo avaliou o uso do fruto da carnaubeira desidratado em 28 cabras no período pós-parto alimentadas com dieta contendo feno de Tifton e concentrado comercial a base de milho (WCWP) ou com o fruto da carnaubeira (CWP). A sincronização de estro foi realizada aos 90 dias pós-parto e todas as fêmeas foram cobertas. No grupo CWP, em relação ao grupo WCWP, o consumo de matéria seca foi significativamente menor (646.48 ± 11.03 g vs. 739.29 ± 3.24 g, respectivamente; P 0.01). O tempo para o aparecimento do primeiro corpo lúteo funcional foi semelhante entre os grupos, ocorrendo em média aos 66 dias pós-parto. No momento da sincronização do estro, o grupo CWP apresentou uma menor espessura do tecido adiposo subcutâneo esternal quando comparado ao grupo WCWP (13,69 ± 0,57 mm vs. 11,93 ± 0,45 mm, respectivamente; P 0,05) e uma menor taxa de gestação (86,67% vs. 46,15%, respectivamente; P 0.02). A taxa de crescimento da vesícula embrionária e do comprimento crânio-caudal, assim como no diâmetro biparietal, torácico e abdominal aos 45 dias de gestação estavam em ambos os grupos dentro do esperado para a espécie em questão. Ao parto a prolificidade não diferiu entre os grupos, com média de 1,39 ± 0,11. Em conclusão, a substituição total do milho pelo fruto da carnaubeira no concentrado fornecido às fêmeas caprinas no período pós-parto não apresentou efeitos...