This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the ... more This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the deleterious effects of doxorubicin (DOXO) in oocytes, follicles and stromal cells in mice ovaries cultured in vitro. In experiment 1, mice ovaries were cultured in DMEM+ alone or supplemented with 5, 50 or 500 ng/mL CIMI, while in experiment 2, mice ovaries were cultured in DMEM+ alone or supplemented with 5 ng/mL CIMI (better concentration), 0.3 μg/mL DOXO or both. Thereafter, the ovaries were processed for histological (morphology, growth, activation, extracellular matrix configuration and stromal cell density), immunohistochemical (caspase-3) analyses. Follicle viability was evaluated by fluorescence microscopy (ethidium homodimer-1 and calcein) while real-time PCR was performed to analyses the levels of (mRNA for SOD, CAT and nuclear factor erythroid 2–related factor 2 (NRF2) analyses. The results showed that DOXO reduces the percentage of normal follicles and the density of stromal ...
SummaryTumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are cytokines that are involve... more SummaryTumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are cytokines that are involved in the development, proliferation and apoptosis of ovarian follicular cells in domestic mammals. The expression of these cytokines in various follicular compartments, depending on the stage of follicle development, demonstrates their involvement in the control of primordial follicle growth up to the preovulatory stage. The mechanism of action of these factors depends on the presence of their receptors that transduce their biological actions. This review shows the expression sites of TNF-α, IL-1β and their receptors in ovarian follicles, and discusses the mechanism of action of these cytokines during follicle development, oocyte maturation and ovulation in domestic animals.
This study evaluates the levels of messenger RNA (mRNA) for eIF4E, PARN, H1FOO, cMOS, GDF9, and C... more This study evaluates the levels of messenger RNA (mRNA) for eIF4E, PARN, H1FOO, cMOS, GDF9, and CCNB1 in oocytes from secondary and antral follicles at different stages of development. The effects of in vitro culture, in vitro prematuration, and in vitro maturation on the expression of these genes on oocytes were also analyzed. The results showed that mRNA levels for H1FOO, GDF9, and PARN were higher in oocytes from small, medium, and large antral follicles, respectively, than those seen in secondary follicles. Oocytes from small, medium, and large antral follicles had higher levels of CCNB1 than oocytes from secondary follicles. Oocytes from cultured secondary follicles had higher levels of GDF9, CMOS, PARN, eIF4E, CCNB1, and H1FOO than before culture. Prematured oocytes from small antral follicles had higher levels of mRNA for GDF9, PARN, and eIF4E than before culture. In addition, higher levels of cMOS and H1FOO were identified in prematured oocytes from medium antral follicles. ...
This study aims to investigate the effect of melatonin on activation, growth and morphology of bo... more This study aims to investigate the effect of melatonin on activation, growth and morphology of bovine primordial follicles, as well as on stromal cells density in ovarian tissues after in vitro culture. Ovarian fragments were cultured in α‐MEM+ alone or supplemented with melatonin (250, 500, 1,000 or 2,000 pM) for a period of six days. Non‐cultured and cultured tissues were processed for histological analysis; according to developmental stages, follicles were classified as primordial or growing follicles. These follicles were further classified as morphologically normal or degenerated. Ovarian stromal cell density was also evaluated. The percentages of primordial and developing follicles, as well as those classified of normal follicles, were compared by Fisher's exact test, and the differences were considered significant when p < .05. The results showed that the presence of 1,000 and 2,000 pM melatonin in culture medium promoted a reduction in the percentage of primordial fol...
This study evaluates the effect of different concentrations (0, 10, 50 and 100ng/mL) of bone morp... more This study evaluates the effect of different concentrations (0, 10, 50 and 100ng/mL) of bone morphogenetic protein-2 (BMP-2) on primordial and secondary follicle development. It also investigates the effects of FSH and BMP-2 on the growth, morphology, ultrastructure and expression of mRNA for GDF9, NLRP5 and NPM2 genes in secondary follicles cultured for 18 days. The presence of BMP-2 at all tested concentrations increased the development of primordial follicles in vitro, but the highest concentration of BMP-2 (100 ng/mL) reduced the percentage of normal follicles when compared with tissues cultured with 10 ng/mL BMP-2. During culture of secondary follicles, in contrast to higher concentrations (50 or 100 ng/mL), 10 ng/mL BMP-2 kept the morphology of follicles during initial stages of in vitro culture. This concentration of BMP-2 also benefits maintenance of the ultrastructure of 18-day cultured follicles. The presence of both BMP-2 and FSH in culture medium resulted in a significan...
Summary This study aims to investigate the effects of jacalin and follicle-stimulating hormone (F... more Summary This study aims to investigate the effects of jacalin and follicle-stimulating hormone (FSH) on activation and survival of goat primordial follicles, as well as on gene expression in cultured ovarian tissue. Ovarian fragments were cultured for 6 days in minimum essential medium (MEM) supplemented with jacalin (10, 25, 50 or 100 μg/ml - Experiment 1) or in MEM supplemented with jacalin (50 μg/ml), FSH (50 ng/ml) or both (Experiment 2). Non-cultured and cultured tissues were processed for histological and ultrastructural analysis. Cultured tissues from Experiment 2 were also stored to evaluate the expression of BMP-15, KL (Kit ligand), c-kit, GDF-9 and proliferating cell nuclear antigen (PCNA) by real-time polymerase chain reaction (PCR). The results of Experiment 1 showed that, compared with tissue that was cultured in control medium, the presence of 50 μg/ml of jacalin increased both the percentages of developing follicles and viability. In Experiment 2, after 6 days, higher...
SummaryThis study investigated mRNA levels for insulin-like growth factors (IGFs) IGF1 (IGF-I) an... more SummaryThis study investigated mRNA levels for insulin-like growth factors (IGFs) IGF1 (IGF-I) and IGF2 (IGF-II), IGF receptors (IGF1R and IGF2R), and binding proteins (IGFBP-1, IGFBP-2. IGFBP-3, IGFBP-4, IGFBP-5 and IGFBP-6) in bovine follicles of 0.2, 0.5 or 1.0 mm in diameter. mRNA expression levels in in vitro cultured follicles that reached approximately 0.5 mm were compared with that of in vivo grown follicles. IGF1R and IGF2R expression levels in 0.5 mm in vivo follicles were higher than in 1.0 or 0.2 mm follicles, respectively. IGFBP-1, IGFBP-2. IGFBP-3, IGFBP-4, IGFBP-5 and IGFBP-6 showed variable expression in the follicular size classes analyzed. In vitro grown follicles had significantly reduced expression levels for IGF1, IGF1R, IGFBP-3, IGFBP-5 and IGFBP-6 mRNA when compared with 0.2 mm follicles, but, when compared with in vivo grown follicles (0.5 mm), only IGFBP-1, IGFBP-2, IGFBP-3 and IGFBP-6 showed a reduction in their expression. In conclusion, IGFs, their recept...
INTRODUÇÃO: A pré-eclâmpsia (PE) é uma desordem complexa que se caracteriza por hipertensão e pro... more INTRODUÇÃO: A pré-eclâmpsia (PE) é uma desordem complexa que se caracteriza por hipertensão e proteinúria após a 20ª semana de gestação, e causa a maior morbidade neonatal e mortalidade perinatal. Até agora, a patogênese exata da PE ainda não é totalmente compreendida, e intensos esforços de pesquisa estão focados na PE para elucidar os mecanismos fisiopatológicos. Estudos apontam que, fatores genéticos estão relacionados à fisiopatologia dessa síndrome. OBJETIVOS: Este estudo objetiva apresentar através de uma revisão de literatura, a relação entre o polimorfismo genético e patogênese da PE, bem como a atuação de alguns genes envolvidos no desenvolvimento da PE. MÉTODOS: Para tanto, realizou-se uma busca nas bases de dados Medline/Pubmed, Lilacs, Biblioteca virtual em saúde (BVS) e Scielo, mediante os descritores: Pré-eclâmpsia; Polimorfismo; e Fatores genéticos, bem como seus correspondentes em inglês. Foram selecionados 14 artigos para compor essa revisão, quem compõe o acervo do...
Despite advances in obstetric medicine, the pathogenesis of preeclampsia (PE) remains poorly unde... more Despite advances in obstetric medicine, the pathogenesis of preeclampsia (PE) remains poorly understood. It has been suggested that PE results from a state of sympathetic hyperactivity with circulating catecholamines increased in this condition. A new enzyme, called Renalase, has recently been identified exhibiting activity on the metabolism of catecholamine and blood pressure reduction when administered in vivo. Thus, this study was conducted to evaluate the possible association between the presence of the Renalase gene (RNLS) (rs10887800) polymorphism and mechanisms that control the pathogenesis of PE. This was a cross-sectional, quantitative, case-control study with 94 pregnant women with PE (cases) and 97 normotensive pregnant women (controls). A standardized form was used to collect demographic and clinical data; oral scraping samples were collected, and DNA extraction and subsequent real-time polymerase chain reaction (PCR) were conducted to evaluate the presence of rs10887800. In terms of genotypic distribution and frequency of alleles, no significant association was observed between the rs10887800 polymorphism and development of PE, or with its severe form. However, the GG genotype was associated with a trend of higher risk of PE (GG vs. AG + AA: OR = 2.16, 0.97-4.86, p = 0.05). Hence, the rs10887800 polymorphism could not be determined as a predisposing factor for PE susceptibility or severity in the studied population.
The aims of the present study were to evaluate the effects of Aloe vera extract on expression of ... more The aims of the present study were to evaluate the effects of Aloe vera extract on expression of mRNA for antioxidant enzymes in bovine ovarian tissue after vitrification, as well as on follicular morphology, viability, activation and extracellular matrix in cultured ovarian tissues that had been previously vitrified. Fragments from bovine ovarian cortical tissue were cryopreserved in a vitrification solution alone or supplemented with two concentrations of Aloe vera (10 or 50%). After thawing, the cryopreserved tissues were analyzed by histological techniques, as well as the levels of mRNA for SOD, CAT, PRDX6 and GPX1 were investigated. Furthermore, cryopreserved fragments were then culture in vitro in α-MEM for 6 days. Histological evaluation of cultured tissues was performed to determine the percentages of normal and developing follicles. The results showed that, after vitrification, the presence of Aloe vera in both concentrations was able to maintain percentages of collagen fibers similar to fresh tissues (P < 0.05). Aloe vera in both concentrations significantly increased mRNA levels for PRDX6 and GPX1 in cryopreserved tissues, while 10% Aloe vera increased mRNA levels for SOD (P < 0.05). In parallel, after in vitro culture, fragments vitrified in the presence of 10% Aloe vera had significantly higher levels of morphologically healthy follicles when compared to tissue that were vitrified without Aloe vera. In fragments vitrified with Aloe vera, the rate of developing follicles was significantly higher than in tissues vitrified without Aloe vera. Tissues vitrified with 10% Aloe vera and cultured in vitro maintained percentages of collagen fibers similar to fresh tissues. In conclusion, 10% Aloe vera increases the expression of mRNA for PRDX6, GPX1 and SOD in vitrified ovarian tissues, maintains follicular survival and promotes activation and development of follicles after in vitro culture of vitrified bovine ovarian tissue.
Summary This study aimed to evaluate the relationship between antral follicular count (AFC) and o... more Summary This study aimed to evaluate the relationship between antral follicular count (AFC) and ovarian volume (OV), preantral follicular population and survival, meiotic progression and ultrastructure of cumulus–oocyte complexes (COCs) after in vitro maturation. In experiment 1, the relationship between AFC and preantral follicle population and survival was evaluated by classical histology. In experiment 2, the relationship among AFC, OV, ability of oocytes to resume meiosis and ultrastructure of in vitro matured bovine COCs was studied. A positive correlation (P < 0.05) between AFC and the numbers of healthy primordial, degenerate and total follicles was observed, as well as with healthy secondary follicles and total follicles. The numbers of grades I and II oocytes in ovaries of high AFC class were higher compared with those with intermediate or lower AFC. After in vitro maturation, COCs from ovaries of high AFC had a higher percentage of oocytes in metaphase II compared with ...
This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the ... more This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the deleterious effects of doxorubicin (DOXO) in oocytes, follicles and stromal cells in mice ovaries cultured in vitro. In experiment 1, mice ovaries were cultured in DMEM+ alone or supplemented with 5, 50 or 500 ng/mL CIMI, while in experiment 2, mice ovaries were cultured in DMEM+ alone or supplemented with 5 ng/mL CIMI (better concentration), 0.3 μg/mL DOXO or both. Thereafter, the ovaries were processed for histological (morphology, growth, activation, extracellular matrix configuration and stromal cell density), immunohistochemical (caspase-3) analyses. Follicle viability was evaluated by fluorescence microscopy (ethidium homodimer-1 and calcein) while real-time PCR was performed to analyses the levels of (mRNA for SOD, CAT and nuclear factor erythroid 2–related factor 2 (NRF2) analyses. The results showed that DOXO reduces the percentage of normal follicles and the density of stromal ...
SummaryTumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are cytokines that are involve... more SummaryTumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are cytokines that are involved in the development, proliferation and apoptosis of ovarian follicular cells in domestic mammals. The expression of these cytokines in various follicular compartments, depending on the stage of follicle development, demonstrates their involvement in the control of primordial follicle growth up to the preovulatory stage. The mechanism of action of these factors depends on the presence of their receptors that transduce their biological actions. This review shows the expression sites of TNF-α, IL-1β and their receptors in ovarian follicles, and discusses the mechanism of action of these cytokines during follicle development, oocyte maturation and ovulation in domestic animals.
This study evaluates the levels of messenger RNA (mRNA) for eIF4E, PARN, H1FOO, cMOS, GDF9, and C... more This study evaluates the levels of messenger RNA (mRNA) for eIF4E, PARN, H1FOO, cMOS, GDF9, and CCNB1 in oocytes from secondary and antral follicles at different stages of development. The effects of in vitro culture, in vitro prematuration, and in vitro maturation on the expression of these genes on oocytes were also analyzed. The results showed that mRNA levels for H1FOO, GDF9, and PARN were higher in oocytes from small, medium, and large antral follicles, respectively, than those seen in secondary follicles. Oocytes from small, medium, and large antral follicles had higher levels of CCNB1 than oocytes from secondary follicles. Oocytes from cultured secondary follicles had higher levels of GDF9, CMOS, PARN, eIF4E, CCNB1, and H1FOO than before culture. Prematured oocytes from small antral follicles had higher levels of mRNA for GDF9, PARN, and eIF4E than before culture. In addition, higher levels of cMOS and H1FOO were identified in prematured oocytes from medium antral follicles. ...
This study aims to investigate the effect of melatonin on activation, growth and morphology of bo... more This study aims to investigate the effect of melatonin on activation, growth and morphology of bovine primordial follicles, as well as on stromal cells density in ovarian tissues after in vitro culture. Ovarian fragments were cultured in α‐MEM+ alone or supplemented with melatonin (250, 500, 1,000 or 2,000 pM) for a period of six days. Non‐cultured and cultured tissues were processed for histological analysis; according to developmental stages, follicles were classified as primordial or growing follicles. These follicles were further classified as morphologically normal or degenerated. Ovarian stromal cell density was also evaluated. The percentages of primordial and developing follicles, as well as those classified of normal follicles, were compared by Fisher's exact test, and the differences were considered significant when p < .05. The results showed that the presence of 1,000 and 2,000 pM melatonin in culture medium promoted a reduction in the percentage of primordial fol...
This study evaluates the effect of different concentrations (0, 10, 50 and 100ng/mL) of bone morp... more This study evaluates the effect of different concentrations (0, 10, 50 and 100ng/mL) of bone morphogenetic protein-2 (BMP-2) on primordial and secondary follicle development. It also investigates the effects of FSH and BMP-2 on the growth, morphology, ultrastructure and expression of mRNA for GDF9, NLRP5 and NPM2 genes in secondary follicles cultured for 18 days. The presence of BMP-2 at all tested concentrations increased the development of primordial follicles in vitro, but the highest concentration of BMP-2 (100 ng/mL) reduced the percentage of normal follicles when compared with tissues cultured with 10 ng/mL BMP-2. During culture of secondary follicles, in contrast to higher concentrations (50 or 100 ng/mL), 10 ng/mL BMP-2 kept the morphology of follicles during initial stages of in vitro culture. This concentration of BMP-2 also benefits maintenance of the ultrastructure of 18-day cultured follicles. The presence of both BMP-2 and FSH in culture medium resulted in a significan...
Summary This study aims to investigate the effects of jacalin and follicle-stimulating hormone (F... more Summary This study aims to investigate the effects of jacalin and follicle-stimulating hormone (FSH) on activation and survival of goat primordial follicles, as well as on gene expression in cultured ovarian tissue. Ovarian fragments were cultured for 6 days in minimum essential medium (MEM) supplemented with jacalin (10, 25, 50 or 100 μg/ml - Experiment 1) or in MEM supplemented with jacalin (50 μg/ml), FSH (50 ng/ml) or both (Experiment 2). Non-cultured and cultured tissues were processed for histological and ultrastructural analysis. Cultured tissues from Experiment 2 were also stored to evaluate the expression of BMP-15, KL (Kit ligand), c-kit, GDF-9 and proliferating cell nuclear antigen (PCNA) by real-time polymerase chain reaction (PCR). The results of Experiment 1 showed that, compared with tissue that was cultured in control medium, the presence of 50 μg/ml of jacalin increased both the percentages of developing follicles and viability. In Experiment 2, after 6 days, higher...
SummaryThis study investigated mRNA levels for insulin-like growth factors (IGFs) IGF1 (IGF-I) an... more SummaryThis study investigated mRNA levels for insulin-like growth factors (IGFs) IGF1 (IGF-I) and IGF2 (IGF-II), IGF receptors (IGF1R and IGF2R), and binding proteins (IGFBP-1, IGFBP-2. IGFBP-3, IGFBP-4, IGFBP-5 and IGFBP-6) in bovine follicles of 0.2, 0.5 or 1.0 mm in diameter. mRNA expression levels in in vitro cultured follicles that reached approximately 0.5 mm were compared with that of in vivo grown follicles. IGF1R and IGF2R expression levels in 0.5 mm in vivo follicles were higher than in 1.0 or 0.2 mm follicles, respectively. IGFBP-1, IGFBP-2. IGFBP-3, IGFBP-4, IGFBP-5 and IGFBP-6 showed variable expression in the follicular size classes analyzed. In vitro grown follicles had significantly reduced expression levels for IGF1, IGF1R, IGFBP-3, IGFBP-5 and IGFBP-6 mRNA when compared with 0.2 mm follicles, but, when compared with in vivo grown follicles (0.5 mm), only IGFBP-1, IGFBP-2, IGFBP-3 and IGFBP-6 showed a reduction in their expression. In conclusion, IGFs, their recept...
INTRODUÇÃO: A pré-eclâmpsia (PE) é uma desordem complexa que se caracteriza por hipertensão e pro... more INTRODUÇÃO: A pré-eclâmpsia (PE) é uma desordem complexa que se caracteriza por hipertensão e proteinúria após a 20ª semana de gestação, e causa a maior morbidade neonatal e mortalidade perinatal. Até agora, a patogênese exata da PE ainda não é totalmente compreendida, e intensos esforços de pesquisa estão focados na PE para elucidar os mecanismos fisiopatológicos. Estudos apontam que, fatores genéticos estão relacionados à fisiopatologia dessa síndrome. OBJETIVOS: Este estudo objetiva apresentar através de uma revisão de literatura, a relação entre o polimorfismo genético e patogênese da PE, bem como a atuação de alguns genes envolvidos no desenvolvimento da PE. MÉTODOS: Para tanto, realizou-se uma busca nas bases de dados Medline/Pubmed, Lilacs, Biblioteca virtual em saúde (BVS) e Scielo, mediante os descritores: Pré-eclâmpsia; Polimorfismo; e Fatores genéticos, bem como seus correspondentes em inglês. Foram selecionados 14 artigos para compor essa revisão, quem compõe o acervo do...
Despite advances in obstetric medicine, the pathogenesis of preeclampsia (PE) remains poorly unde... more Despite advances in obstetric medicine, the pathogenesis of preeclampsia (PE) remains poorly understood. It has been suggested that PE results from a state of sympathetic hyperactivity with circulating catecholamines increased in this condition. A new enzyme, called Renalase, has recently been identified exhibiting activity on the metabolism of catecholamine and blood pressure reduction when administered in vivo. Thus, this study was conducted to evaluate the possible association between the presence of the Renalase gene (RNLS) (rs10887800) polymorphism and mechanisms that control the pathogenesis of PE. This was a cross-sectional, quantitative, case-control study with 94 pregnant women with PE (cases) and 97 normotensive pregnant women (controls). A standardized form was used to collect demographic and clinical data; oral scraping samples were collected, and DNA extraction and subsequent real-time polymerase chain reaction (PCR) were conducted to evaluate the presence of rs10887800. In terms of genotypic distribution and frequency of alleles, no significant association was observed between the rs10887800 polymorphism and development of PE, or with its severe form. However, the GG genotype was associated with a trend of higher risk of PE (GG vs. AG + AA: OR = 2.16, 0.97-4.86, p = 0.05). Hence, the rs10887800 polymorphism could not be determined as a predisposing factor for PE susceptibility or severity in the studied population.
The aims of the present study were to evaluate the effects of Aloe vera extract on expression of ... more The aims of the present study were to evaluate the effects of Aloe vera extract on expression of mRNA for antioxidant enzymes in bovine ovarian tissue after vitrification, as well as on follicular morphology, viability, activation and extracellular matrix in cultured ovarian tissues that had been previously vitrified. Fragments from bovine ovarian cortical tissue were cryopreserved in a vitrification solution alone or supplemented with two concentrations of Aloe vera (10 or 50%). After thawing, the cryopreserved tissues were analyzed by histological techniques, as well as the levels of mRNA for SOD, CAT, PRDX6 and GPX1 were investigated. Furthermore, cryopreserved fragments were then culture in vitro in α-MEM for 6 days. Histological evaluation of cultured tissues was performed to determine the percentages of normal and developing follicles. The results showed that, after vitrification, the presence of Aloe vera in both concentrations was able to maintain percentages of collagen fibers similar to fresh tissues (P < 0.05). Aloe vera in both concentrations significantly increased mRNA levels for PRDX6 and GPX1 in cryopreserved tissues, while 10% Aloe vera increased mRNA levels for SOD (P < 0.05). In parallel, after in vitro culture, fragments vitrified in the presence of 10% Aloe vera had significantly higher levels of morphologically healthy follicles when compared to tissue that were vitrified without Aloe vera. In fragments vitrified with Aloe vera, the rate of developing follicles was significantly higher than in tissues vitrified without Aloe vera. Tissues vitrified with 10% Aloe vera and cultured in vitro maintained percentages of collagen fibers similar to fresh tissues. In conclusion, 10% Aloe vera increases the expression of mRNA for PRDX6, GPX1 and SOD in vitrified ovarian tissues, maintains follicular survival and promotes activation and development of follicles after in vitro culture of vitrified bovine ovarian tissue.
Summary This study aimed to evaluate the relationship between antral follicular count (AFC) and o... more Summary This study aimed to evaluate the relationship between antral follicular count (AFC) and ovarian volume (OV), preantral follicular population and survival, meiotic progression and ultrastructure of cumulus–oocyte complexes (COCs) after in vitro maturation. In experiment 1, the relationship between AFC and preantral follicle population and survival was evaluated by classical histology. In experiment 2, the relationship among AFC, OV, ability of oocytes to resume meiosis and ultrastructure of in vitro matured bovine COCs was studied. A positive correlation (P < 0.05) between AFC and the numbers of healthy primordial, degenerate and total follicles was observed, as well as with healthy secondary follicles and total follicles. The numbers of grades I and II oocytes in ovaries of high AFC class were higher compared with those with intermediate or lower AFC. After in vitro maturation, COCs from ovaries of high AFC had a higher percentage of oocytes in metaphase II compared with ...
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