Effects of BMP-4 and FSH on growth, morphology and mRNA expression of oocyte-secreted factors in cultured bovine secondary follicles

This study evaluated the effect of different concentrations of bone morphogenetic protein-4 (BMP-4), as well as the interaction of BMP-4 and follicle stimulating hormone (FSH) on growth, ultrastructural integrity, and expression of mRNA for growth differentiation factor-9 (GDF-9), BMP-15, maternal antigen that the embryo requires (Mater) and nucleoplasmin-2 (Npm-2) in bovine secondary follicles cultured in vitro for 18 days. Follicles cultured in the presence of 50 ng/ml BMP-4 had a progressive increase in their diameters with the increase of culture period from 0 to 6 and 12 days, but no significant differences were observed among treatments. The presence of both FSH and BMP-4 in a culture medium did not stimulate follicle growth when compared to the control medium. After 12 days, the percentage of normal follicles was maintained similar to that of day 0 in the medium supplemented with both FSH and BMP-4, but no significant differences among treatments were observed after 18 days of culture. BMP-4 maintained the ultrastructural integrity of follicles after 18 days of culture, while follicles cultured in medium supplemented with FSH or both BMP-4 and FSH had oocyte with irregular zona pellucida, vesicular bodies, and an abundance of vacuoles. Follicles cultured in the presence of BMP-4 had an increase in the levels of BMP-15 mRNA, when compared to those cultured in medium supplemented with FSH alone. In conclusion, the addition of BMP-4 in culture medium contributes to preserve follicular ultrastructure, but BMP-4 did not interact positively with FSH. Regarding secondary follicles cultured in the presence of FSH, BMP-4 increases the expression of mRNA for BMP-15.(AU)

Effects of BMP-4 and FSH on growth, morphology and mRNA expression of oocyte-secreted factors in cultured bovine secondary follicles

This study evaluated the effect of different concentrations of bone morphogenetic protein-4 (BMP-4), as well as the interaction of BMP-4 and follicle stimulating hormone (FSH) on growth, ultrastructural integrity, and expression of mRNA for growth differentiation factor-9 (GDF-9), BMP-15, maternal antigen that the embryo requires (Mater) and nucleoplasmin-2 (Npm-2) in bovine secondary follicles cultured in vitro for 18 days. Follicles cultured in the presence of 50 ng/ml BMP-4 had a progressive increase in their diameters with the increase of culture period from 0 to 6 and 12 days, but no significant differences were observed among treatments. The presence of both FSH and BMP-4 in a culture medium did not stimulate follicle growth when compared to the control medium. After 12 days, the percentage of normal follicles was maintained similar to that of day 0 in the medium supplemented with both FSH and BMP-4, but no significant differences among treatments were observed after 18 days of culture. BMP-4 maintained the ultrastructural integrity of follicles after 18 days of culture, while follicles cultured in medium supplemented with FSH or both BMP-4 and FSH had oocyte with irregular zona pellucida, vesicular bodies, and an abundance of vacuoles. Follicles cultured in the presence of BMP-4 had an increase in the levels of BMP-15 mRNA, when compared to those cultured in medium supplemented with FSH alone. In conclusion, the addition of BMP-4 in culture medium contributes to preserve follicular ultrastructure, but BMP-4 did not interact positively with FSH. Regarding secondary follicles cultured in the presence of FSH, BMP-4 increases the expression of mRNA for BMP-15.

Proteínas morfogenéticas ósseas (BMPs) e seu papel na regulação da oogênese e da foliculogênese ovariana em mamíferos / The bone morphogenetic proteins (BMPs) and its role in the regulation of ovarian folliculogenesis and oogenesis in mammalian

O processo de foliculogênese é controlado por uma variedade de gonadotrofinas e de fatores de crescimento locais, que agem em conjunto para regular a formação e o desenvolvimento dos folículos ovarianos. Dentre esses fatores, destacam-se as proteínas morfogenéticas ósseas (BMPs), que representam uma família de fatores de crescimento amplamente estudados e que se caracterizam por controlar as funções ovarianas em diferentes estágios do desenvolvimento folicular. Dessa forma, a presente revisão tem como foco principal descrever os locais de expressão das BMPs dos tipos 2, 4, 6, 7 e 15 e discutir o papel delas, bem como das gonadotrofinas FSH e LH durante a foliculogênese em mamíferos.(AU)

The process of folliculogenesis is controlled by a variety of gonadotropins and local growth factors that act together to regulate formation and development of ovarian follicles. Among these factors, the bone morphogenetic proteins (BMPs) represent a family of growth factors widely studied that have important functions at different stages of ovarian follicular development. Thus, this review aims to describe the local of expression and to discuss the role of BMPs 2, 4, 6, 7 and 15 as well as the gonadotropins FSH and LH during folliculogenesis in mammals. (AU)

Proteínas morfogenéticas ósseas (BMPs) e seu papel na regulação da oogênese e da foliculogênese ovariana em mamíferos / The bone morphogenetic proteins (BMPs) and its role in the regulation of ovarian folliculogenesis and oogenesis in mammalian

O processo de foliculogênese é controlado por uma variedade de gonadotrofinas e de fatores de crescimento locais, que agem em conjunto para regular a formação e o desenvolvimento dos folículos ovarianos. Dentre esses fatores, destacam-se as proteínas morfogenéticas ósseas (BMPs), que representam uma família de fatores de crescimento amplamente estudados e que se caracterizam por controlar as funções ovarianas em diferentes estágios do desenvolvimento folicular. Dessa forma, a presente revisão tem como foco principal descrever os locais de expressão das BMPs dos tipos 2, 4, 6, 7 e 15 e discutir o papel delas, bem como das gonadotrofinas FSH e LH durante a foliculogênese em mamíferos.

The process of folliculogenesis is controlled by a variety of gonadotropins and local growth factors that act together to regulate formation and development of ovarian follicles. Among these factors, the bone morphogenetic proteins (BMPs) represent a family of growth factors widely studied that have important functions at different stages of ovarian follicular development. Thus, this review aims to describe the local of expression and to discuss the role of BMPs 2, 4, 6, 7 and 15 as well as the gonadotropins FSH and LH during folliculogenesis in mammals.

FSH and LH enhance the development of goat preantral follicles cultured in vitro

This study evaluated th e effect of increased follicle stimulating hormone (FSH) concentrations on the expression of mRNA for LH receptors after in vitro culture of goat preantral follicles ( ≥ 150 μm) for 18 days. It also investigated whether the addition of luteinizing hormone (LH) to the culture medium, which contained increasing concentrations of FSH throughout the culture period, influenced the surv ival, growth and antrum formation of in vitro cultured goat preantral follicles. In experiment 1, preantral follicles were cultured in α -MEM + or α -MEM + supplemented with increasing concentrations of FSH throughout the culture period (sequential medium: FSH 100 ng/ml (days 0 to 6), FSH 500 ng/ml (days 6 to 12) and FSH 1000 ng/ml (days 12 to 18). The expression of luteinizing hormone receptor (LHR) was analyzed in noncultured and cultured follicles using real time RT-PCR. In experiment 2, isolated preantral follicles were cultured for 18 days in a sequential medium containing FSH (control) or a control medium supplemented with LH (50 or 100 ng/ml) from day 12 of culture onwards. Follicle development was evaluated on the basis of antr al cavity formation as well as follicular and oocyte growth after in vitro maturation. FSH stimulated a significant increase in the expression of mRNA for LH receptors after 18 days of culture. Furthermore, after 18 days, all tested media promoted follicular survival and antrum formation; however, a significant increase in the rate of follicular growth and resumption of meiosis was ob served when LH was used compared to the control. In conclusion, preantral follicles cultured in a medium supplemented with FSH increased LH receptor mR NA levels. Moreover, the addition of LH to the culture medium containing increasing concentrations of FSH (sequential medium) improved the in vitro development of goat preantral follicles.

FSH and LH enhance the development of goat preantral follicles cultured in vitro

This study evaluated th e effect of increased follicle stimulating hormone (FSH) concentrations on the expression of mRNA for LH receptors after in vitro culture of goat preantral follicles ( ≥ 150 μm) for 18 days. It also investigated whether the addition of luteinizing hormone (LH) to the culture medium, which contained increasing concentrations of FSH throughout the culture period, influenced the surv ival, growth and antrum formation of in vitro cultured goat preantral follicles. In experiment 1, preantral follicles were cultured in α -MEM + or α -MEM + supplemented with increasing concentrations of FSH throughout the culture period (sequential medium: FSH 100 ng/ml (days 0 to 6), FSH 500 ng/ml (days 6 to 12) and FSH 1000 ng/ml (days 12 to 18). The expression of luteinizing hormone receptor (LHR) was analyzed in noncultured and cultured follicles using real time RT-PCR. In experiment 2, isolated preantral follicles were cultured for 18 days in a sequential medium containing FSH (control) or a control medium supplemented with LH (50 or 100 ng/ml) from day 12 of culture onwards. Follicle development was evaluated on the basis of antr al cavity formation as well as follicular and oocyte growth after in vitro maturation. FSH stimulated a significant increase in the expression of mRNA for LH receptors after 18 days of culture. Furthermore, after 18 days, all tested media promoted follicular survival and antrum formation; however, a significant increase in the rate of follicular growth and resumption of meiosis was ob served when LH was used compared to the control. In conclusion, preantral follicles cultured in a medium supplemented with FSH increased LH receptor mR NA levels. Moreover, the addition of LH to the culture medium containing increasing concentrations of FSH (sequential medium) improved the in vitro development of goat preantral follicles.(AU)

Effects of BMP-7 and FSH on the development of goat preantral follicles and levels of mRNA for FSH-R, BMP-7 and BMP receptors after in-vitro culture

This study aimed to evaluate the effects of FSH and BMP - 7 on growth and on expression of FSH - R , BMP - 7 and BMP receptors in cultured secondary follicles. Goat secondary follicles (~200 μm) were isolated and cultured in vitro, with 5% CO 2 in air at 39°C, for 6 days in the presence of BMP - 7 (50 ng/m l ) supplemented or not with FSH (50 ng/m l ). Follicular diameter and the formation of the antrum were evaluated before and after culture. For each treatment, at the end of culture period, groups of 6 follicles were collected and, after extraction of total RNA and cDNA synthesis, the levels of mRNA for FSH - R, BMP - 7 and BMP receptors in cultured secondary follicles were quantified by real time PCR. The results showed that addition of BMP - 7 or F SH to culture medium stimulated growth of secondary follicles , while addition of both BMP - 7 and FSH was needed to significantly increase the percentage of follicles forming an antrum and the follicular levels of mRNA for both BMP - 7 and FSH - R. For BMP receptors, FSH reduced the levels of mRNA for BMPR - IA and BMP - RII in comparison with those follicles cultured in MEM alone and supplemented with BMP - 7, respectively. In conclusion , l ike FSH, BMP - 7 affect s in vitro growth of cultured secondary follicle s , but it stimulates antrum formation and expression of the mRNA ’s for BMP - 7 and FSH - R only in presence of both BMP - 7 and FSH . However, the levels of mRNA f or BMP - RIA and BMP - RII are reduced in follicles culture d in medium supplemented with FSH.

Effects of BMP-7 and FSH on the development of goat preantral follicles and levels of mRNA for FSH-R, BMP-7 and BMP receptors after in-vitro culture

This study aimed to evaluate the effects of FSH and BMP - 7 on growth and on expression of FSH - R , BMP - 7 and BMP receptors in cultured secondary follicles. Goat secondary follicles (~200 μm) were isolated and cultured in vitro, with 5% CO 2 in air at 39°C, for 6 days in the presence of BMP - 7 (50 ng/m l ) supplemented or not with FSH (50 ng/m l ). Follicular diameter and the formation of the antrum were evaluated before and after culture. For each treatment, at the end of culture period, groups of 6 follicles were collected and, after extraction of total RNA and cDNA synthesis, the levels of mRNA for FSH - R, BMP - 7 and BMP receptors in cultured secondary follicles were quantified by real time PCR. The results showed that addition of BMP - 7 or F SH to culture medium stimulated growth of secondary follicles , while addition of both BMP - 7 and FSH was needed to significantly increase the percentage of follicles forming an antrum and the follicular levels of mRNA for both BMP - 7 and FSH - R. For BMP receptors, FSH reduced the levels of mRNA for BMPR - IA and BMP - RII in comparison with those follicles cultured in MEM alone and supplemented with BMP - 7, respectively. In conclusion , l ike FSH, BMP - 7 affect s in vitro growth of cultured secondary follicle s , but it stimulates antrum formation and expression of the mRNA s for BMP - 7 and FSH - R only in presence of both BMP - 7 and FSH . However, the levels of mRNA f or BMP - RIA and BMP - RII are reduced in follicles culture d in medium supplemented with FSH.(AU)