Effects of FTO and PPARγ variants on intrauterine growth restriction in a Brazilian birth cohort.

Intrauterine growth restriction (IUGR) is related to a higher risk of neonatal mortality, minor cognitive deficit, metabolic syndrome, and cardiovascular disease in adulthood. In previous studies, genetic variants in the FTO (fat mass and obesity-associated) and PPARγ (peroxisome proliferator-activated receptor-gamma) genes have been associated with metabolic disease, body mass index, and obesity among other outcomes. We studied the association of selected FTO (rs1421085, rs55682395, rs17817449, rs8043757, rs9926289, and rs9939609) and PPARγ (rs10865710, rs17036263, rs35206526, rs1801282, rs28763894, rs41516544, rs62243567, rs3856806, and rs1805151) single-nucleotide polymorphisms (SNPs) with IUGR, through a case-control study in a cohort of live births that occurred from June 1978 to May 1979 in a Brazilian city. We selected 280 IUGR cases and 256 controls for analysis. Logistic regression was used to jointly analyze the SNPs as well as factors such as maternal smoking, age, and schooling. We found that the PPARγ rs41516544 increased the risk of IUGR for male offspring (OR 27.83, 95%CI 3.65-212.32) as well as for female offspring (OR=8.94, 95%CI: 1.96-40.88). The FTO rs9939609 TA genotype resulted in a reduced susceptibility to IUGR for male offspring only (OR=0.47, 95%CI: 0.26-0.86). In conclusion, we demonstrated that PPARγ SNP had a positive effect and FTO SNP had a negative effect on IUGR occurrence, and these effects were gender-specific.

Is soft drink consumption associated with gestational hypertension? Results from the BRISA cohort.

It is still unknown whether excessive consumption of sugar-sweetened beverages may be linked to gestational hypertensive disorders, other than preeclampsia. This study investigated the association between soft drink consumption and hypertension during pregnancy, analyzing the relationship from the perspective of counterfactual causal theory. Data from pregnant women of the BRISA cohort were analyzed (1,380 in São Luis and 1,370 in Ribeirão Preto, Brazil). The explanatory variable was the frequency of soft drink consumption during pregnancy obtained in a prenatal interview. The outcome was gestational hypertension based on medical diagnosis, at the time of delivery. A theoretical model of the association between soft drink consumption and gestational hypertension was constructed using a directed acyclic graph. Marginal structural models (MSM) weighted by the inverse of the probability of soft drink consumption were also employed. Using Poisson regression analysis, high soft drink consumption (≥7 times/week) was associated with gestational hypertension in São Luís (RR=1.48; 95%CI: 1.03-2.10), in Ribeirão Preto (RR=1.51; 95%CI: 1.13-2.01), and in the two cohorts combined (RR=1.45; 95%CI: 1.16-1.82) compared to lower exposure (<7 times/week). In the MSM, the association between high soft drink consumption and gestational hypertension was observed in Ribeirão Preto (RR=1.63; 95%CI: 1.21-2.19) and in the two cohorts combined (RR=1.51; 95%CI: 1.15-1.97), but not in São Luís (RR=1.26; 95%CI: 0.79-2.00). High soft drink consumption seems to be a risk factor for gestational hypertension, suggesting that it should be discouraged during pregnancy.

Racial inequality in perinatal outcomes in two Brazilian birth cohorts.

This study aimed to estimate and compare racial inequality in low birth weight (LBW), preterm birth (PTB), and intrauterine growth restriction (IUGR) in two Brazilian birth cohorts. This was a cross-sectional study nested within two birth cohorts in Ribeirão Preto (RP) and São Luís (SL), whose mothers were interviewed from January to December 2010. In all, 7430 (RP) and 4995 (SL) mothers were interviewed. The maternal skin color was the exposure variable. Associations were adjusted for socioeconomic and biological covariates: maternal education, per capita family income, family economic classification, household head occupation, maternal age, parity, marital status, prenatal care, type of delivery, maternal pre-pregnancy BMI, hypertension, hypertension during pregnancy, and smoking during pregnancy collected from questionnaires applied at birth. Statistical analysis was done with the chi-squared test and logistic regression. In RP, newborns from mothers with black skin color had a higher risk of LBW and IUGR, even after adjusting for socioeconomic and biological variables (P<0.001). In SL, skin color was not a risk factor for LBW (P=0.859), PTB (P=0.220), and IUGR (P=0.062), before or after adjustment for socioeconomic and biological variables. The detection of racial inequality in these perinatal outcomes only in the RP cohort after adjustment for socioeconomic and biological factors may be reflecting the existence of racial discrimination in the RP society. In contrast, the greater miscegenation present in São Luís may be reflecting less racial discrimination of black and brown women in this city.

Is soft drink consumption associated with gestational hypertension? Results from the BRISA cohort

It is still unknown whether excessive consumption of sugar-sweetened beverages may be linked to gestational hypertensive disorders, other than preeclampsia. This study investigated the association between soft drink consumption and hypertension during pregnancy, analyzing the relationship from the perspective of counterfactual causal theory. Data from pregnant women of the BRISA cohort were analyzed (1,380 in São Luis and 1,370 in Ribeirão Preto, Brazil). The explanatory variable was the frequency of soft drink consumption during pregnancy obtained in a prenatal interview. The outcome was gestational hypertension based on medical diagnosis, at the time of delivery. A theoretical model of the association between soft drink consumption and gestational hypertension was constructed using a directed acyclic graph. Marginal structural models (MSM) weighted by the inverse of the probability of soft drink consumption were also employed. Using Poisson regression analysis, high soft drink consumption (≥7 times/week) was associated with gestational hypertension in São Luís (RR=1.48; 95%CI: 1.03-2.10), in Ribeirão Preto (RR=1.51; 95%CI: 1.13-2.01), and in the two cohorts combined (RR=1.45; 95%CI: 1.16-1.82) compared to lower exposure (<7 times/week). In the MSM, the association between high soft drink consumption and gestational hypertension was observed in Ribeirão Preto (RR=1.63; 95%CI: 1.21-2.19) and in the two cohorts combined (RR=1.51; 95%CI: 1.15-1.97), but not in São Luís (RR=1.26; 95%CI: 0.79-2.00). High soft drink consumption seems to be a risk factor for gestational hypertension, suggesting that it should be discouraged during pregnancy.

Racial inequality in perinatal outcomes in two Brazilian birth cohorts

This study aimed to estimate and compare racial inequality in low birth weight (LBW), preterm birth (PTB), and intrauterine growth restriction (IUGR) in two Brazilian birth cohorts. This was a cross-sectional study nested within two birth cohorts in Ribeirão Preto (RP) and São Luís (SL), whose mothers were interviewed from January to December 2010. In all, 7430 (RP) and 4995 (SL) mothers were interviewed. The maternal skin color was the exposure variable. Associations were adjusted for socioeconomic and biological covariates: maternal education, per capita family income, family economic classification, household head occupation, maternal age, parity, marital status, prenatal care, type of delivery, maternal pre-pregnancy BMI, hypertension, hypertension during pregnancy, and smoking during pregnancy collected from questionnaires applied at birth. Statistical analysis was done with the chi-squared test and logistic regression. In RP, newborns from mothers with black skin color had a higher risk of LBW and IUGR, even after adjusting for socioeconomic and biological variables (P<0.001). In SL, skin color was not a risk factor for LBW (P=0.859), PTB (P=0.220), and IUGR (P=0.062), before or after adjustment for socioeconomic and biological variables. The detection of racial inequality in these perinatal outcomes only in the RP cohort after adjustment for socioeconomic and biological factors may be reflecting the existence of racial discrimination in the RP society. In contrast, the greater miscegenation present in São Luís may be reflecting less racial discrimination of black and brown women in this city.

Effects of FTO and PPARγ variants on intrauterine growth restriction in a Brazilian birth cohort

Intrauterine growth restriction (IUGR) is related to a higher risk of neonatal mortality, minor cognitive deficit, metabolic syndrome, and cardiovascular disease in adulthood. In previous studies, genetic variants in the FTO (fat mass and obesity-associated) and PPARγ (peroxisome proliferator-activated receptor-gamma) genes have been associated with metabolic disease, body mass index, and obesity among other outcomes. We studied the association of selected FTO (rs1421085, rs55682395, rs17817449, rs8043757, rs9926289, and rs9939609) and PPARγ (rs10865710, rs17036263, rs35206526, rs1801282, rs28763894, rs41516544, rs62243567, rs3856806, and rs1805151) single-nucleotide polymorphisms (SNPs) with IUGR, through a case-control study in a cohort of live births that occurred from June 1978 to May 1979 in a Brazilian city. We selected 280 IUGR cases and 256 controls for analysis. Logistic regression was used to jointly analyze the SNPs as well as factors such as maternal smoking, age, and schooling. We found that the PPARγ rs41516544 increased the risk of IUGR for male offspring (OR 27.83, 95%CI 3.65-212.32) as well as for female offspring (OR=8.94, 95%CI: 1.96-40.88). The FTO rs9939609 TA genotype resulted in a reduced susceptibility to IUGR for male offspring only (OR=0.47, 95%CI: 0.26-0.86). In conclusion, we demonstrated that PPARγ SNP had a positive effect and FTO SNP had a negative effect on IUGR occurrence, and these effects were gender-specific.

Socio-environmental determinants of the delay in the first dental visit: results of two population-based cohort studies in Brazil

The objective of this study was to describe the timing of the first dental visit and investigate the association of socioeconomic and behavioral factors with dental visit delay among 10/11-year-old children from two live-birth population cohorts with extremely contrasting socioeconomic profiles. Follow-up data (2004-2005) from cohorts of Ribeirão Preto (RP) (n=790) and São Luís (SL) (n=673) were evaluated. Delay in dental visit was defined as not visiting a dentist before the age of 7. Covariates included family socioeconomic characteristics, mother-related health behavior, and child-related characteristics. Prevalence ratios with robust standard errors were estimated. In both cohorts, less than 5% of children had visited a dentist before the age of two and about 35% of them had not visited a dentist before the age of seven. Lower mother's schooling and lack of private health insurance were associated with the delay in first dental visit for both cohorts. A small number of mother's prenatal care visits and being from a single-father family or a family without parents were only associated in the RP cohort, while having ≥4 siblings and lifetime dental pain were associated in the SL cohort. The association with dental pain probably reveals a preventive care-seeking behavior. Therefore, the percentage of delayed first dental visit of children was very high even among those with the most educated mothers. Further studies are necessary to analyze recent changes and underlying factors related to access to first dental visit after the implementation of the National Oral Health Policy in 2006.

Body image dissatisfaction and symptoms of depression disorder in adolescents

The objective of this study was to evaluate the effect of body image dissatisfaction on symptoms of depressive disorder in adolescents. This is a cross-sectional study that included 2,162 adolescents ages 18-19 born in São Luís, Maranhão, Brazil, which was part of the joint RPS cohort (Brazilian birth cohorts of Ribeirão Preto-SP, Pelotas-RS, and São Luís-MA). Socioeconomic characteristics, nutritional status, mental health, and body image characteristics were evaluated. Body image was assessed by Stunkard's silhouettes scale. The presence of symptoms indicative of depressive disorder was investigated through a diagnostic interview MINI (Mini International Neuropsychiatric Interview). A theoretical model was built in a Directed Acyclic Graph (DAG) in order to investigate the relationship between the variables of the study. The relationship was estimated weighting the inverse probability of selection for the variables of adjustment: sex and nutritional status. Among the dissatisfied adolescents due to overweight, 66.54% were girls, 32.85% were overweight, and 11.99% were obese (P<0.01). There was a significant association between dissatisfaction due to overweight and symptoms of depressive disorder (P=0.01), and there was no evidence of the same association with dissatisfaction due to thinness. Therefore, only dissatisfaction due to overweight was associated with the symptoms of depressive disorder in the evaluated adolescents.

Relative validity of a food frequency questionnaire for adolescents from a capital in the Northeastern region of Brazil

The present study was conducted to evaluate the validity of the Food Frequency Questionnaire (FFQ) used in the RPS Birth Cohort Consortium (Ribeirão Preto, Pelotas, and São Luís) to assess dietary intake of adolescents from São Luís, Maranhão. The research was developed with 152 adolescents aged 18 and 19 years. For the validation of the FFQ, the average of three 24-hour recalls (24HRs) was used as the reference method. The mean and standard deviation of energy and nutrient intake extracted from the surveys were estimated. The paired Student's t-test was used to verify the differences between the instruments. Pearson correlation coefficient, intraclass correlation coefficient (ICC), weighted Kappa, and the Bland-Altman plot were calculated in order to measure the agreement. The study adopted a level of significance <5%. Compared with the three 24HRs, the FFQ overestimated the consumption of most nutrients. Energy-adjusted and de-attenuated concordance Pearson correlation coefficients ranged from 0.06 to 0.43, and correlations were significant for iron, calcium, riboflavin, sodium, saturated fat, niacin, and vitamin C. The energy-adjusted and de-attenuated ICCs ranged from 0.01 to 0.31, and the weighted Kappa ranged from 0.01 to 0.46. The analyses of agreement were significant for vitamin C, fiber, calcium, riboflavin, niacin, sodium, lipids, and iron. In conclusion, the FFQ presented acceptable relative validity for lipids, saturated fatty acids, fiber, calcium, iron, riboflavin, niacin, vitamin C, and sodium. This instrument will be useful in studies about food consumption of adolescents in São Luís, Maranhão.

Relative validity of a food frequency questionnaire for adolescents from a capital in the Northeastern region of Brazil.

The present study was conducted to evaluate the validity of the Food Frequency Questionnaire (FFQ) used in the RPS Birth Cohort Consortium (Ribeirão Preto, Pelotas, and São Luís) to assess dietary intake of adolescents from São Luís, Maranhão. The research was developed with 152 adolescents aged 18 and 19 years. For the validation of the FFQ, the average of three 24-hour recalls (24HRs) was used as the reference method. The mean and standard deviation of energy and nutrient intake extracted from the surveys were estimated. The paired Student's t-test was used to verify the differences between the instruments. Pearson correlation coefficient, intraclass correlation coefficient (ICC), weighted Kappa, and the Bland-Altman plot were calculated in order to measure the agreement. The study adopted a level of significance <5%. Compared with the three 24HRs, the FFQ overestimated the consumption of most nutrients. Energy-adjusted and de-attenuated concordance Pearson correlation coefficients ranged from 0.06 to 0.43, and correlations were significant for iron, calcium, riboflavin, sodium, saturated fat, niacin, and vitamin C. The energy-adjusted and de-attenuated ICCs ranged from 0.01 to 0.31, and the weighted Kappa ranged from 0.01 to 0.46. The analyses of agreement were significant for vitamin C, fiber, calcium, riboflavin, niacin, sodium, lipids, and iron. In conclusion, the FFQ presented acceptable relative validity for lipids, saturated fatty acids, fiber, calcium, iron, riboflavin, niacin, vitamin C, and sodium. This instrument will be useful in studies about food consumption of adolescents in São Luís, Maranhão.

Socio-environmental determinants of the delay in the first dental visit: results of two population-based cohort studies in Brazil.

The objective of this study was to describe the timing of the first dental visit and investigate the association of socioeconomic and behavioral factors with dental visit delay among 10/11-year-old children from two live-birth population cohorts with extremely contrasting socioeconomic profiles. Follow-up data (2004-2005) from cohorts of Ribeirão Preto (RP) (n=790) and São Luís (SL) (n=673) were evaluated. Delay in dental visit was defined as not visiting a dentist before the age of 7. Covariates included family socioeconomic characteristics, mother-related health behavior, and child-related characteristics. Prevalence ratios with robust standard errors were estimated. In both cohorts, less than 5% of children had visited a dentist before the age of two and about 35% of them had not visited a dentist before the age of seven. Lower mother's schooling and lack of private health insurance were associated with the delay in first dental visit for both cohorts. A small number of mother's prenatal care visits and being from a single-father family or a family without parents were only associated in the RP cohort, while having ≥4 siblings and lifetime dental pain were associated in the SL cohort. The association with dental pain probably reveals a preventive care-seeking behavior. Therefore, the percentage of delayed first dental visit of children was very high even among those with the most educated mothers. Further studies are necessary to analyze recent changes and underlying factors related to access to first dental visit after the implementation of the National Oral Health Policy in 2006.

Body image dissatisfaction and symptoms of depression disorder in adolescents.

The objective of this study was to evaluate the effect of body image dissatisfaction on symptoms of depressive disorder in adolescents. This is a cross-sectional study that included 2,162 adolescents ages 18-19 born in São Luís, Maranhão, Brazil, which was part of the joint RPS cohort (Brazilian birth cohorts of Ribeirão Preto-SP, Pelotas-RS, and São Luís-MA). Socioeconomic characteristics, nutritional status, mental health, and body image characteristics were evaluated. Body image was assessed by Stunkard's silhouettes scale. The presence of symptoms indicative of depressive disorder was investigated through a diagnostic interview MINI (Mini International Neuropsychiatric Interview). A theoretical model was built in a Directed Acyclic Graph (DAG) in order to investigate the relationship between the variables of the study. The relationship was estimated weighting the inverse probability of selection for the variables of adjustment: sex and nutritional status. Among the dissatisfied adolescents due to overweight, 66.54% were girls, 32.85% were overweight, and 11.99% were obese (P<0.01). There was a significant association between dissatisfaction due to overweight and symptoms of depressive disorder (P=0.01), and there was no evidence of the same association with dissatisfaction due to thinness. Therefore, only dissatisfaction due to overweight was associated with the symptoms of depressive disorder in the evaluated adolescents.

Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos

Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.

Características das carcaças e carne de tourinhos Nelore alimentados com níveis de concentrado e farelo do mesocarpo de babaçu / Carcass and meat characteristics of cattle fed concentrate levels and babassu mesocarp bran

Avaliaram-se as características da carcaça e da carne de tourinhos Nelore alimentados com diferentes níveis de concentrado e farelo do mesocarpo do babaçu na dieta. Foram utilizados 28 tourinhos da raça Nelore, com idade inicial de 21 meses e 356,66±19,25kg de peso corporal inicial, distribuídos inteiramente ao acaso em dietas com dois níveis de concentrado (65 e 71%), associadas ou não à inclusão de 35% de farelo do mesocarpo do babaçu no concentrado. A elevação da fração concentrada na dieta de 65 para 71%, bem como a inclusão de 35% de farelo do mesocarpo do babaçu no concentrado, não alterou a quantidade de músculo, gordura e osso da carcaça, com valores médios de 63,91; 19,46 e 16,63% do peso de carcaça fria, respectivamente. O rendimento dos cortes comerciais secundários do traseiro especial não foi alterado pela inclusão de 35% de farelo do mesocarpo do babaçu no concentrado nem pelo teor de concentrado da dieta. A força ao cisalhamento das fibras musculares, a perda de líquido por descongelamento e cozimento não foram alteradas pelos fatores estudados, apresentando valores médios de 6,57kgf/cm3, 16,08% e 22,20%, respectivamente. O marmoreio aumentou (5,78 para 9,79 pontos) pelo avanço do nível de concentrado na dieta, mas não foi alterado pela inclusão do farelo do mesocarpo do babaçu. O farelo do mesocarpo do babaçu apresenta características nutricionais que possibilitam substituir parcialmente o milho em dietas com elevada proporção de concentrado, não alterando a composição física da carcaça e a qualidade da carne de tourinhos Nelore confinados.

Carcass characteristics and meat of young bulls fed different levels of concentrate and babassu mesocarp bran in the diet were evaluated. A total of 28 Nellore young bulls, initial age of 21 months and 356.66±19.25kg of initial body weight were randomized in diets containing two levels of concentrate (65% and 71) associated or not to the inclusion of 35% of babassu mesocarp bran in the concentrate. The elevation of the concentrate fraction of the diet from 65 to 71%, and the inclusion of 35% of babassu mesocarp bran in the concentrate did not alter the amount of muscle, fat and bone of the carcass, with average values of 63.91, 19.46 and 16.63% of cold carcass weight, respectively. The yields of retail cuts from the pistol cut were not altered by the concentrate level of the diet or by the inclusion of 35% babassu mesocarp bran in the concentrate. The shear force of muscle fibers and the losses of liquid during thawing and cooking were not altered by the factors studied, with average values of 6.57kgf/cm3, 16.06% and 22.20%, respectively. Marbling increased (5.78 to 9.79 points) to advance the level of concentrate in the diet, but was not altered by the inclusion of babassu mesocarp bran. Babassu mesocarp bran has nutritional characteristics that allow partial replacement of corn in diets with high concentrate, not changing physical carcass composition and meat quality of confined young Nellore bulls.

Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos.

Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.

Effects of angiotensin II, atrial natriuretic peptide and endothelin-1 on proliferation and steroidogenic output of bovine granulosa cells cultured in a chemically defined system.

The role of local factors in the modulation of granulosa cell (GC) proliferation and differentiation is well described in the literature. The present work used a long-term bovine GC culture, in chemically defined medium without gonadotropins, to study the effects of angiotensin II (Ang II), atrial natriuretic peptide (ANP) and endothelin-1 (EDN1) on the steroidogenesis and cellular proliferation. Small follicles (3-5mm in diameter) from ovaries obtained in the slaughterhouse were selected according to their vascularization and follicular fluid color in order to isolate GC. Granulosa cells were plated at a density of 5×10(4)cells/well in supplemented alpha-MEM containing 3 levels (0, 10(-8)M and 10(-7)M) of Ang II, ANP, and EDN1 for up to 96h. Proliferation was evaluated by tritiated thymidine incorporation. The results showed that Ang II, ANP, and EDN1 modulate the steroidogenic output and proliferation index of GCs depending on the dose and time of culture. The selected vasoactive peptides increased androstenedione (A4) consumption in parallel with increased estradiol (E2). Although the peptides also promoted a significant increase in pregnenolone (P5) and progesterone (P4) production, the E2:P4 ratio was maintained at a high at most of the tested doses. Taken together, our in vitro data suggest that these vasoactive factors may have a direct effect on physiological follicular deviation, favoring dominance of the selected follicle.

Effects of maternal separation on the dietary preference and behavioral satiety sequence in rats.

This study investigated the effects of maternal separation on the feeding behavior of rats. A maternal separation model was used on postnatal day 1 (PND1), forming the following groups: in the maternal separation (MS) group, pups were separated from their mothers each day from PND1 to PND14, whereas in the control (C) group pups were kept with their mothers. Subgroups were formed to study the effects of light and darkness: control with dark and light exposure, female and male (CF and CM), and maternal separation with dark and light exposure, female and male (SDF, SDM, SLF and SLM). Female rats had higher caloric intake relative to body weight compared with male controls in the dark period only (CF=23.3±0.5 v. CM=18.2±0.7, P<0.001). Macronutrient feeding preferences were observed, with male rats exhibiting higher caloric intake from a protein diet as compared with female rats (CF=4.1±0.7, n=8 v. CM=7.0±0.5, n=8, P<0.05) and satiety development was not interrupted. Female rats had a higher adrenal weight as compared with male rats independently of experimental groups and exhibited a higher concentration of serum triglycerides (n=8, P<0.001). The study indicates possible phenotypic adjustments in the structure of feeding behavior promoted by maternal separation, especially in the dark cycle. The dissociation between the mother's presence and milk intake probably induces adjustments in feeding behavior during adulthood.

Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions.

Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17ß-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.

Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions

Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.