RESUMEN
BACKGROUND: This study investigated the psychocultural perspectives concerning family quality of life among Brazilian families with children who have severe or profound intellectual disability. METHODS: Individual in-depth semi-structured interviews conducted with 15 mothers, selected by convenience, were analysed using a categorical thematic analysis technique. The themes were examined to allow for an interpretative approach of the results. RESULTS: Mothers revealed that their children with disabilities had insufficient access to services and support related to health care, transportation and recreation. Family quality of life was negatively affected by financial restrictions and social interaction difficulties. Caring for a child with disabilities seemed to be centred on the mother and religious coping appeared as a common psychological adjustment strategy. CONCLUSIONS: Improving emotional and psychological cares, as well as social and practical measures comprising income support and access to appropriate health care, were inferred to be the mothers' priorities to improve their families' quality of life.
Asunto(s)
Adaptación Psicológica , Familia/psicología , Accesibilidad a los Servicios de Salud , Discapacidad Intelectual/enfermería , Calidad de Vida/psicología , Adulto , Brasil/etnología , Niño , Estudios Transversales , Familia/etnología , Femenino , Humanos , Discapacidad Intelectual/etnología , Masculino , Persona de Mediana Edad , Madres/psicología , Investigación Cualitativa , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Primordial follicles, the main source of oocytes in the ovary, are essential for the maintenance of fertility throughout the reproductive lifespan. To the best of our knowledge, there are no reports describing the effect of anethole on this important ovarian follicle population. The aim of the study was to investigate the effect of different anethole concentrations on the in vitro culture of caprine preantral follicles enclosed in ovarian tissue. Randomized ovarian fragments were fixed immediately (non-cultured treatment) or distributed into five treatments: α-MEM+ (cultured control), α-MEM+ supplemented with ascorbic acid at 50 µg/mL (AA), and anethole at 30 (AN30), 300 (AN300), or 2000 µg/mL (AN2000), for 1 or 7 days. After 7 days of culture, a significantly higher percentage of morphologically normal follicles was observed when anethole at 2000 µg/mL was used. For both culture times, a greater percentage of growing follicles was observed with the AN30 treatment compared to AA and AN2000 treatments. Anethole at 30 and 2000 µg/mL concentrations at days 1 and 7 of culture resulted in significantly larger follicular diameter than in the cultured control treatment. Anethole at 30 µg/mL concentration at day 7 showed significantly greater oocyte diameter than the other treatments, except when compared to the AN2000 treatment. At day 7 of culture, levels of reactive oxygen species (ROS) were significantly lower in the AN30 treatment than the other treatments. In conclusion, supplementation of culture medium with anethole improves survival and early follicle development at different concentrations in the caprine species.
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Anisoles/farmacología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Folículo Ovárico/crecimiento & desarrollo , Estrés Oxidativo/efectos de los fármacos , Derivados de Alilbenceno , Animales , Anisoles/administración & dosificación , Medios de Cultivo , Relación Dosis-Respuesta a Droga , Femenino , Cabras , Inmunohistoquímica , Técnicas de Maduración In Vitro de los Oocitos/métodos , Folículo Ovárico/efectos de los fármacos , Distribución AleatoriaRESUMEN
Primordial follicles, the main source of oocytes in the ovary, are essential for the maintenance of fertility throughout the reproductive lifespan. To the best of our knowledge, there are no reports describing the effect of anethole on this important ovarian follicle population. The aim of the study was to investigate the effect of different anethole concentrations on the in vitro culture of caprine preantral follicles enclosed in ovarian tissue. Randomized ovarian fragments were fixed immediately (non-cultured treatment) or distributed into five treatments: α-MEM+ (cultured control), α-MEM+ supplemented with ascorbic acid at 50 μg/mL (AA), and anethole at 30 (AN30), 300 (AN300), or 2000 µg/mL (AN2000), for 1 or 7 days. After 7 days of culture, a significantly higher percentage of morphologically normal follicles was observed when anethole at 2000 μg/mL was used. For both culture times, a greater percentage of growing follicles was observed with the AN30 treatment compared to AA and AN2000 treatments. Anethole at 30 and 2000 µg/mL concentrations at days 1 and 7 of culture resulted in significantly larger follicular diameter than in the cultured control treatment. Anethole at 30 µg/mL concentration at day 7 showed significantly greater oocyte diameter than the other treatments, except when compared to the AN2000 treatment. At day 7 of culture, levels of reactive oxygen species (ROS) were significantly lower in the AN30 treatment than the other treatments. In conclusion, supplementation of culture medium with anethole improves survival and early follicle development at different concentrations in the caprine species.
Asunto(s)
Animales , Femenino , Estrés Oxidativo/efectos de los fármacos , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Folículo Ovárico/crecimiento & desarrollo , Anisoles/farmacología , Cabras , Inmunohistoquímica , Distribución Aleatoria , Medios de Cultivo , Relación Dosis-Respuesta a Droga , Técnicas de Maduración In Vitro de los Oocitos/métodos , Folículo Ovárico/efectos de los fármacos , Anisoles/administración & dosificaciónRESUMEN
The effects of epidermal growth factor (EGF) concentrations (0, 10, 50, and 100ng/ml) on in vitro culture (IVC) of equine preantral follicles were evaluated using histology, estradiol and reactive oxygen species (ROS) production and metabolomics. After IVC, the percentage of normal follicles was lower (P<0.05) for all treatments when compared to non-cultured control. EGF 50ng/ml treatment had more (P<0.05) normal follicles at Day 7 of culture when compared with EGF 0 and 100ng/ml. EGF 50ng/ml had more (P<0.05) developing follicles than the 0ng/ml and 10ng/ml EGF treatments. Follicular and oocyte diameters were greater (P<0.05) with EGF 50ng/ml than the other cultured treatments, but similar (P>0.05) to the non-cultured control. From Day 1 to Day 7 estradiol production increased (P<0.05) in all EGF treatments. EGF 50ng/ml was the only treatment that maintained ROS production through IVC. Metabolomics profiles of the spent media indicated that eleven ions from variable influence in the projection (VIP) scores were higher represented in the EGF 50ng/ml treatment. In conclusion, EGF 50ng/ml treatment maintained follicle survival and ROS production, and promoted activation of cultured equine preantral follicles enclosed in ovarian tissue.
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Factor de Crecimiento Epidérmico/farmacología , Caballos , Metabolómica , Folículo Ovárico/efectos de los fármacos , Técnicas de Cultivo de Tejidos/veterinaria , Animales , Medios de Cultivo/química , Medios de Cultivo/farmacología , Relación Dosis-Respuesta a Droga , Factor de Crecimiento Epidérmico/química , Estradiol/metabolismo , Femenino , Oocitos/metabolismo , Folículo Ovárico/fisiología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Our aim has been to evaluate the effect of cryoprotective agents (CPAs) on the exposure, vitrification (VIT), and in vitro culture (IVC) of ovarian tissue with regard to the expression and immunolocalization of aquaporins (AQPs) 3 and 9 in ovine preantral follicles. Tissues were treated as follows: Experiment I: (1) control (without exposure to CPAs), (2) e-EG (exposure to ethylene glycol), (3) er-EG (exposure to and removal of EG), (4) e-DMSO (exposure to dimethyl sulfoxide), (5) er-DMSO (exposure to and removal of DMSO), (6) e-EG+DMSO (exposure to EG+DMSO), (7) er-EG+DMSO (exposure to and removal of EG+DMSO); Experiment II: (1) control, (2) VIT, (3) IVC, (4) VIT-IVC. In Experiment I, following er-EG or er-DMSO, tissue showed the down-regulation (P < 0.05) of AQP3 mRNA. The mRNA transcript levels were reduced (P < 0.05) for AQP9 in tissue following er-EG+DMSO. Immunolocalization was positive for both proteins (AQP3 and AQP9) on ovine preantral follicles following all treatments, except in the e-EG+DMSO group. In Experiment II, the mRNA levels of AQP3 and AQP9 following VIT treatment were similar (P > 0.05) to that of the control group. Nevertheless, VIT-IVC treatment led to the down-regulation of mRNA of AQP3 and AQP9. Thus, AQP3 and AQP9 act in a mutually dependent way, maintaining the cell homeostasis that is essential for the ovary cryopreservation process. Furthermore, the changes in the expression profiles of mRNA and protein after culture are a strong indicator that in vitro conditions have to be strictly controlled to ensure follicle viability and functionality.
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Acuaporinas/metabolismo , Crioprotectores/farmacología , Ovario/metabolismo , Oveja Doméstica/metabolismo , Técnicas de Cultivo de Tejidos , Animales , Acuaporinas/genética , Femenino , Inmunohistoquímica , Folículo Ovárico/citología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Vitrificación/efectos de los fármacosRESUMEN
This study investigated the effect of insulin concentration on the in vitro culture of equine preantral follicles enclosed in ovarian tissue. Ovarian tissue samples were immediately fixed (noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0 ng/mL, 10 ng/mL, or 10 µg/mL insulin. Ovarian tissues were processed and analyzed by classical histology. Culture medium samples were collected after 1 and 7 days of culture for steroid and reactive oxygen species (ROS) analyses. The percentage of morphologically normal follicles was greater (P < 0.001) in insulin-treated groups after 1 day of culture; likewise, more (P < 0.02) normal follicles were observed after 7 days of culture in medium supplemented with 10-ng/mL insulin. Furthermore, an increase (P < 0.01) in developing (transition, primary, and secondary) follicles between Days 1 and 7 of culture was observed only with the 10-ng/mL insulin treatment. ROS production after 1 or 7 days of culture was lower (P < 0.0001) in medium with 10-ng/mL insulin than the other treatments. Ovarian tissues containing preantral follicles were able to produce estradiol and progesterone after 1 and 7 days of culture; however, treatments did not differ in steroid production. In conclusion, the use of a physiological concentration (10 ng/mL) of insulin rather than the previously reported concentration (10 µg/mL) for in vitro culture of equine preantral follicles improved follicular survival and growth and lowered oxidative stress. Results from this study shed light on new perspectives for producing an appropriate medium to improve equine preantral follicle in vitro survival and growth.
Asunto(s)
Caballos , Insulina/farmacología , Folículo Ovárico/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Técnicas de Cultivo de Tejidos/veterinaria , Animales , Medios de Cultivo , Femenino , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Técnicas Reproductivas Asistidas/veterinariaRESUMEN
This study investigated the effect of adding different concentrations of bovine recombinant follicle-stimulating hormone on the IVC of equine preantral follicles enclosed in ovarian tissue fragments. Randomized ovarian fragments were fixed immediately (fresh noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0, 10, 50, and 100 ng/mL FSH and subsequently analyzed by classical histology. Culture media collected on Day 1 or Day 7 and were analyzed for steroids (estradiol and progesterone) and reactive oxygen species (ROS). After Day 1 and Day 7 of culture, 50-ng/mL FSH treatment had a greater (P < 0.05) percentage of morphologically normal follicles when compared to the other groups, except the 10-ng/mL FSH treatment at Day 1 of culture. The percentage of developing follicles (transition, primary, and secondary), and follicular and oocyte diameters were higher (P < 0.05) in the 50-ng/mL FSH treatment compared to the other groups after Day 7 of culture. Furthermore, estradiol secretion and ROS production were maintained (P > 0.05) throughout the culture in the 50-ng/mL FSH treatment. In conclusion, the addition of 50 ng/mL of FSH promoted activation of primordial follicles to developing follicles, improved survival of preantral follicles, and maintained estradiol and ROS production of equine ovarian tissue after 7 days of culture.
Asunto(s)
Hormona Folículo Estimulante/farmacología , Caballos , Folículo Ovárico/efectos de los fármacos , Animales , Medios de Cultivo , Estradiol/metabolismo , Femenino , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Progesterona/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Técnicas Reproductivas Asistidas/veterinaria , Técnicas de Cultivo de Tejidos/veterinariaRESUMEN
The aims of this study were to verify the steady-state level of vasoactive intestinal peptide (VIP) mRNA in goat follicles at various developmental stages and to investigate the influence of VIP on the survival, antrum formation and growth of secondary follicles cultured for 6 days. Primordial, primary and secondary goat follicles and small and large antral follicles were obtained to quantify VIP mRNA by real-time reverse transcription with the polymerase chain reaction. The influence of VIP and the presence or absence of follicle-stimulating hormone (FSH) on the development of secondary follicles and on mRNA expression for VIP and FSH receptor (FSHR) were determined after 6 days of culture. Survival, antrum formation and follicular diameter were evaluated every other day of culture. The levels of VIP mRNA in primary and secondary follicles were significantly higher than in primordial follicles. Cumulusoocyte complexes (COCs) from both small and large antral follicles had significantly higher levels ofVIP mRNA than their respective granulosa/theca cells. During culture, the addition of VIP and/or FSH had o effect on follicular development. However, the presence of FSH and/or VIP in the culture medium significantly reduced VIP mRNA levels, but did not alter FSHR mRNA levels. In conclusion, VIP mRNA was detected in all goat follicular categories and cellular types, VIP and/or FSH did not affect the development of secondary follicles and reduce the expression of VIP mRNA levels.
Asunto(s)
Femenino , Animales , Folículo Ovárico , Ovario , Péptido Intestinal Vasoactivo , ARN Mensajero , Rumiantes , Hormona Folículo EstimulanteRESUMEN
The aims of this study were to verify the steady-state level of vasoactive intestinal peptide (VIP) mRNA in goat follicles at various developmental stages and to investigate the influence of VIP on the survival, antrum formation and growth of secondary follicles cultured for 6 days. Primordial, primary and secondary goat follicles and small and large antral follicles were obtained to quantify VIP mRNA by real-time reverse transcription with the polymerase chain reaction. The influence of VIP and the presence or absence of follicle-stimulating hormone (FSH) on the development of secondary follicles and on mRNA expression for VIP and FSH receptor (FSHR) were determined after 6 days of culture. Survival, antrum formation and follicular diameter were evaluated every other day of culture. The levels of VIP mRNA in primary and secondary follicles were significantly higher than in primordial follicles. Cumulusoocyte complexes (COCs) from both small and large antral follicles had significantly higher levels ofVIP mRNA than their respective granulosa/theca cells. During culture, the addition of VIP and/or FSH had o effect on follicular development. However, the presence of FSH and/or VIP in the culture medium significantly reduced VIP mRNA levels, but did not alter FSHR mRNA levels. In conclusion, VIP mRNA was detected in all goat follicular categories and cellular types, VIP and/or FSH did not affect the development of secondary follicles and reduce the expression of VIP mRNA levels.(AU)
Asunto(s)
Animales , Femenino , ARN Mensajero , Péptido Intestinal Vasoactivo , Ovario , Folículo Ovárico , Rumiantes , Hormona Folículo EstimulanteRESUMEN
The aim of this study was to evaluate the effect of vascular endothelial growth factor-A(165) (VEGF-A(165)) on the in vitro development of goat secondary preantral follicles. Preantral follicles (≥150 µm in diameter) were isolated from the ovaries of adult mixed-breed goats and individually cultured for 18 days in αMEM in the absence (control) or presence of VEGF-A(165) at concentrations of 10 ng/ml (VEGF10) and 100 ng/ml (VEGF100). Analyses of follicular survival, diameter, antrum formation and rate of daily growth were performed every 6 days. At the end of the culture period, morphologically normal oocytes (≥110 µm in diameter) were taken for in vitro maturation (IVM). The results demonstrated that all follicles presented oocytes and granulosa cells that were morphologically normal and after labeling with calcein-AM, high rates of oocyte viability were observed in all treatments. The follicular diameter and the growth rate achieved in the presence of VEGF10 were higher than those of the control. Both treatments with VEGF-A(165) showed higher rates of oocyte recovery for IVM when compared with the control. Moreover, only the addition of VEGF-A(165) permitted oocytes grown in vitro to reach metaphase II. Thus, the addition of VEGF-A(165) to the culture medium improves the development of goat preantral follicles cultured in vitro, allowing the production of mature oocytes.
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Oocitos/citología , Oocitos/efectos de los fármacos , Folículo Ovárico/citología , Folículo Ovárico/crecimiento & desarrollo , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cromatina/metabolismo , Femenino , Cabras , Humanos , Meiosis/efectos de los fármacos , Oocitos/metabolismo , Folículo Ovárico/efectos de los fármacosRESUMEN
This study investigates steady-state level of bone morphogenetic protein-15 (BMP-15) mRNA in caprine follicles, and the effects of BMP-15 on in vitro development of preantral follicles. Ovarian fragments were cultured for one or seven days in Minimal Essential Medium (MEM(+)) with BMP-15 (0, 1, 10, 50, 100 or 200 ng/mL), and further analyzed by histology, transmission electron and fluorescent microscopy. BMP-15 mRNA in secondary follicles was higher than in primordial and primary follicles. After seven days, 10, 50 or 100 ng/mL of BMP-15 maintained the percentage of normal follicles similar to the control (non-cultured), and increased the oocyte and follicle diameters when compared to the control and MEM(+). BMP-15 at 100 ng/mL increased the secondary follicles and maintained their ultrastructural integrity. In conclusion, the BMP-15 mRNAs were detected in all follicular categories. BMP-15 (100 ng/mL) maintained the integrity and promoted the growth of caprine preantral follicles cultured for seven days.
Asunto(s)
Proteína Morfogenética Ósea 15/metabolismo , Cabras/crecimiento & desarrollo , Folículo Ovárico/crecimiento & desarrollo , Animales , Proteína Morfogenética Ósea 15/genética , Proteína Morfogenética Ósea 15/farmacología , Supervivencia Celular/efectos de los fármacos , Femenino , Microscopía Electrónica de Transmisión , Oocitos/efectos de los fármacos , Oocitos/crecimiento & desarrollo , Folículo Ovárico/anatomía & histología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/metabolismo , Técnicas de Cultivo de Tejidos , Transcripción GenéticaRESUMEN
This study evaluated the expression of FSH receptors (FSHR) in the different stages of goat follicle development and investigated whether the addition of increasing concentrations of FSH throughout the culture period influences the survival, growth and antral formation of in vitro-cultured caprine preantral follicles. The expression of FSHR was analysed before and after culturing follicles using real-time RT-PCR. For the culture, preantral follicles (≥150 µm) were isolated from ovarian fragments and cultured for 18 days in α-MEM+ alone or associated with recombinant FSH (rFSH: 100 or 1000 ng/ml), or in α-MEM+ supplemented with increasing concentrations of FSH throughout culture periods as follows: (a) sequential medium 1: FSH 100 ng/ml (from day 0 to 6), FSH 500 ng/ml (from day 6 to 12) and FSH 1000 ng/ml (from day 12 to 18); and (b) sequential medium 2: FSH 500 ng/ml (from day 0 to 9) and 1000 ng/ml (from day 9 to 18). Follicle development was evaluated on the basis of antral cavity formation, follicular and oocyte growth, and cumulus-oocyte complex health. The expression of FSHR in isolated caprine follicles increased from the preantral to antral phase. Regarding the culture, after 18 days, sequential medium 1 promoted follicular survival, antrum formation and a reduction in oocyte extrusion. Both sequential media promoted a higher rate of meiotic resumption compared with the other treatments. In conclusion, the addition of increased concentrations of FSH (sequential medium) has a significant impact on the in vitro development of caprine preantral follicles.
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Folículo Ovárico/citología , Folículo Ovárico/fisiología , Receptores de HFE/genética , Animales , Células Cultivadas , Medios de Cultivo , Femenino , Hormona Folículo Estimulante/farmacología , Cabras , Hormonas/farmacología , Técnicas In Vitro , Oocitos/citología , Oocitos/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , ARN Mensajero/genética , Receptores de HFE/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The aim of the present study was to evaluate the effects of different medium replacement intervals on the viability, antral cavity formation, growth and in vitro maturation (IVM) of oocytes from caprine and ovine pre-antral follicles. Pre-antral ovarian follicles (≥ 150 µm) were isolated from the ovarian cortex of goats and sheep and were individually cultured for 24 days using two different medium replacement intervals [2 days (T(1) ) or 6 days (T(2) )]. Follicle development was evaluated on the basis of antral cavity formation, increases in follicular diameter and the presence of healthy cumulus oocyte complexes and fully grown oocytes. For caprine species, results showed a higher percentage (p<0.05) of viable follicles in T(1) than T(2) from day 6 until the end of the culture. In addition, when comparing both treatments after the same culture duration, the rate of antrum formation was significantly higher in T(1) than in T(2) from day 12 onwards. Yet, in ovines, when both treatments were compared on day 24 of the culture, there were more viable follicles in T(2) than in T(1) (p<0.05). In the caprine species, percentages of fully grown oocytes (≥ 110 µm) acceptable for IVM after 24 days of culture were significantly higher in normal follicles cultured in T(1) (30.0%) than in T(2) (6.7%; p<0.05). On the other hand, in ovines, at the end of the culture, the percentage of oocytes destined for IVM was higher in T(2) than in T(1) (23.5% vs 2.9%; p<0.05). In conclusion, under the same conditions, the frequency of medium replacement significantly affected the in vitro development of caprine and ovine pre-antral follicles. To improve the efficiency of the culture system, the medium must be replaced every 2 and 6 days for goat and sheep pre-antral follicles, respectively.
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Medios de Cultivo , Cabras , Folículo Ovárico/fisiología , Ovinos , Animales , Femenino , Meiosis , Oocitos/citología , Folículo Ovárico/citología , Folículo Ovárico/crecimiento & desarrollo , Especificidad de la Especie , Técnicas de Cultivo de Tejidos/métodos , Técnicas de Cultivo de Tejidos/veterinariaRESUMEN
This study aims to investigate the effects of follicle stimulating hormone (FSH) and fibroblast growth factor-2 (FGF-2) on the survival and growth of caprine preantral follicles. Ovarian tissues were cultured for 1, 7, 14, 21 or 28 days in medium supplemented with FSH (FSH-2d or FSH-7d, i.e., with replacement of the culture medium every 2 or 7 days, respectively) or FSH+FGF-2 (replacement of the medium every 2 days). Non-cultured (control) and cultured ovarian fragments were processed for histological and ultrastructural analysis. After 28 days of culture, the media supplemented with FSH-2d was the most effective in maintaining the percentage of normal follicles and in promoting follicular growth. Furthermore, both treatments with FSH increased the percentage of the primary follicles. However, ultrastructural studies did not confirm follicular integrity from 14 days of culture onward. In conclusion, culturing tissue for up to 7 days in medium containing FSH alone or combined with FGF-2 maintains caprine preantral follicle integrity and promotes their growth in vitro.
Asunto(s)
Cabras/metabolismo , Folículo Ovárico/crecimiento & desarrollo , Animales , Medios de Cultivo , Femenino , Factor 2 de Crecimiento de Fibroblastos/farmacología , Hormona Folículo Estimulante/farmacología , Microscopía Electrónica de Transmisión/veterinaria , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/ultraestructura , Factores de Tiempo , Técnicas de Cultivo de Tejidos/veterinariaRESUMEN
In addition to the central nervous system, vasoactive intestinal peptide (VIP) containing nerves have been described throughout the female genital tract. VIP is reported to be produced by nerves fibers innervating follicles at all stages of development in rodents. There is growing evidence that VIP and their receptors play important roles in the local regulation of ovarian physiology mostly through cAMP pathway. It has been reported that VIP regulates the ovarian follicle survival and growth, oocyte maturation, ovulation and steroidogenesis. Studies also demonstrated that VIP inhibits apoptosis of rat follicles and is an important factor for the growth of preantral follicles enclosed in caprine ovarian tissue. Even though the addition of VIP to the culture medium did not improve in vitro maturation and fertilization of oocytes, it has been shown to stimulate ovulation in perfused rat ovaries. VIP is also involved in the regulation of steroidogenic activity. Therefore, this review aims to summarize current data on the importance of VIP on ovarian physiology.
Asunto(s)
Animales , Ovulación/fisiología , Oocitos/fisiología , Sistema Nervioso Central/anatomía & histologíaRESUMEN
In addition to the central nervous system, vasoactive intestinal peptide (VIP) containing nerves have been described throughout the female genital tract. VIP is reported to be produced by nerves fibers innervating follicles at all stages of development in rodents. There is growing evidence that VIP and their receptors play important roles in the local regulation of ovarian physiology mostly through cAMP pathway. It has been reported that VIP regulates the ovarian follicle survival and growth, oocyte maturation, ovulation and steroidogenesis. Studies also demonstrated that VIP inhibits apoptosis of rat follicles and is an important factor for the growth of preantral follicles enclosed in caprine ovarian tissue. Even though the addition of VIP to the culture medium did not improve in vitro maturation and fertilization of oocytes, it has been shown to stimulate ovulation in perfused rat ovaries. VIP is also involved in the regulation of steroidogenic activity. Therefore, this review aims to summarize current data on the importance of VIP on ovarian physiology.(AU)
Asunto(s)
Animales , Ovulación/fisiología , Oocitos/fisiología , Sistema Nervioso Central/anatomía & histologíaRESUMEN
The aim of this study was to develop a dynamic culture medium containing FSH, LH and EGF to promote the in vitro development of oocytes obtained from goat preantral follicles to complete maturation and to improve the capacity of these oocytes for in vitro fertilization (IVF) and embryo production. For experiment I, preantral follicles were cultured for 18 days in medium supplemented with increasing concentrations of FSH (T1 - control) or in control medium added LH alone or in association with EGF: T2 (LH 50 ng/ml), T3 (LH 50 ng/ml + EGF 50 ng/ml), T4 (LH 50 ng/ml + EGF 100 ng/ml), T5 (LH 100 ng/ml), T6 (LH 100 ng/ml + EGF 50 ng/ml) and T7 (LH 100 ng/ml + EGF 100 ng/ml). For experiment II, preantral follicles were cultured only in the culture medium used in T7, and after 18 days, their oocytes underwent in vitro maturation (IVM) followed by IVF. At the end of the culture period, T3, T4 and T7 had a positive influence on the daily follicular growth rate. Oocytes grown in T4 and T7 had a meiosis resumption percentage significantly superior to the other treatments. Two embryos were obtained, in which preantral follicles in medium supplemented with 100 ng/ml LH and 100 ng/ml EGF (T7). In conclusion, our sequential culture system was able to promote the in vitro growth of preantral follicles, promoting their oocyte maturation and caprine embryo production from preantral follicles.
Asunto(s)
Medios de Cultivo , Fertilización In Vitro/veterinaria , Cabras/embriología , Oocitos/fisiología , Folículo Ovárico/citología , Folículo Ovárico/fisiología , Animales , Células Cultivadas , Factor de Crecimiento Epidérmico/administración & dosificación , Femenino , Hormona Folículo Estimulante/administración & dosificación , Hormona Luteinizante/administración & dosificación , Meiosis , Oocitos/citología , Técnicas de Cultivo de Tejidos/veterinariaRESUMEN
The aim of this study was to evaluate the effect of vasoactive intestinal peptide (VIP)on the survival, activation and growth of goat preantral follicles after in vitro culture. The ovarian cortex was divided into small pieces and one fragment was immediately fixed (control). The remaining fragments were cultured in vitro for 1 or 7 days at 39 degrees C and 5% CO(2), in supplemented minimum essential medium (MEM(+)) with or without different concentrations of VIP (1, 10, 50, 100 or 200 ng/ml). Noncultured (fresh control) and cultured ovarian fragments were processed for histological analysis and transmission electron microscopy. Follicles were classified as primordial or developing, and as normal or degenerated. Our findings indicate that when compared with control, addition of all concentrations of VIP except 200 ng/ml resulted in similar percentages of normal preantral follicles after 1 and 7 days of culture. Culture of ovarian cortex tissue for 1 and 7 days increased the percentage of follicular activation in all treatments when compared with control, except with 1 ng/ml of VIP after 1 day. However, no difference was observed between VIP-treated and MEM(+)-treated follicles. In addition, after 7 days of culture, the highest follicular and oocyte diameters were observed in follicles cultured with 10 ng/ml VIP relative to MEM(+) alone. Transmission electron microscopy showed ultrastructural integrity of follicles after 7 days of culture in 10 ng/ml VIP. In conclusion, this study demonstrates that VIP maintains follicular integrity and stimulates caprine preantral follicle growth.
Asunto(s)
Folículo Ovárico/fisiología , Péptido Intestinal Vasoactivo/farmacología , Animales , Medios de Cultivo/farmacología , Femenino , Cabras , Técnicas de Cultivo de Tejidos , Péptido Intestinal Vasoactivo/administración & dosificaciónRESUMEN
The aim of this study was to investigate the effects of estradiol and follicle-stimulating hormone (FSH) on survival and growth of caprine preantral follicles. Pieces of ovarian tissue were cultured for 1 or 7 days in minimum essential medium (MEM) containing estradiol (1, 5, 10, 20 or 40 pg/ml), FSH (50 ng/ml), or a combination of the two hormones. Cultured and noncultured control ovarian tissues were processed for histological and ultrastructural studies. The results showed that after 7 days of culture, the treatments that yielded the highest percentage of normal follicles relative to MEM alone were those that combined FSH with estradiol at 1, 5 or 20 pg/ml. The addition of FSH to 1-day cultures containing 1 pg/ml estradiol or to 7-day cultures with 1 or 5 pg/ml estradiol increased the percentage of normal follicles compared to estradiol alone at the same concentrations. After 7 days of culture, all treatments generated higher percentages of developing follicles as compared to control and MEM alone. The addition of either FSH or 10 pg/ml of estradiol to the culture media or estradiol (1, 5, 10 or 20 pg/ml) and FSH in combination significantly increased follicular diameter as compared with MEM alone following 7 days of culture. Ultrastructural studies confirmed follicular integrity after 7 days of culture in the presence of 1 pg/ml estradiol plus FSH. In conclusion, this study demonstrated that the interaction between estradiol and FSH maintains ultrastructural integrity and stimulates activation and further growth of cultured caprine preantral follicles.
Asunto(s)
Estradiol/administración & dosificación , Hormona Folículo Estimulante/administración & dosificación , Hormonas/administración & dosificación , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/crecimiento & desarrollo , Animales , Técnicas de Cultivo de Célula , Supervivencia Celular , Medios de Cultivo/química , Relación Dosis-Respuesta a Droga , Estradiol/química , Femenino , Hormona Folículo Estimulante/química , Cabras , Hormonas/química , Microscopía Electrónica de Transmisión , Folículo Ovárico/ultraestructura , Factores de TiempoRESUMEN
This study investigated the effects of androstenedione and follicle-stimulating hormone (FSH) on the viability and growth of caprine preantral follicles. Ovarian tissues were cultured for 1 or 7 days in Minimum. Essential Medium (MEM) containing androstenedione (0, 1, 10, 50 or 100 ng/ml). FSH (50 ng/ml), or a combination of these two hormones. Cultured and non-cultured control tissues were processed for histological and fluorescence analysis. In comparison with non-cultured control, a sinfnificant reduction was noted in the percentage of normal follicles in all treatments after 1 and 7 days of culture (except in all treatment with 1 ng/ml of androstenedione for 1 day). As the culture period progressed from 1 to 7 days, treatments with 10 ng/ml of androstenedione + FSH or 50 ng/ml of androstenedione alone maintained the percentage of normal follicles. After 1 day, treatments with 10, 50, or 100 ng/ml of androstenedione + FSH, or with 50 ng/ml of androstenedione alone had more developing follicles than fresh control tissue. When comparing the culture periods, treatments with 1, 10 or 100 ng/ml of androstenedione alone, or FSH alone, or FSH with 1 ng/ml of androstenedione, showed an increase in the percentage of developing follicles. After 1 and 7 days, there were no differences in oocyte and follicular diameter among the treated samples and non-cultured control or MEM+ cultured tissue. Fluorescence analysis demonstrated that only fragments cultured in 50 or 100 ng/ml of androstenedione + FSH had viable preantral follicles similar to those observed in MEM+ alone. In conclusion, androstenedione at 50 or 100 ng/ml, either associated with FSH or at 50 ng/ml alone, plays an important role in the maintenance of caprine preantral follicle viability and activation after only a short in vitro culture period. In addition, after 7 days MEM+ alone was efficient in the maintenance of viability and in follicular activation, showing the importance of basic medium composition.