212 resultados para Blastocyst
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Myosins are molecular motors associated with the actin cytoskeleton that participate in the mechanisms of cellular motility. During the development of the nervous system, migration of nerve cells to specific sites, extension of growth cones, and axonal transport are dramatic manifestations of cellular motility. We demonstrate, via immunoblots, the expression of myosin Va during early stages of embryonic development in chicks, extending from the blastocyst period to the beginning of the fetal period. The expression of myosin Va in specific regions and cellular structures of the nervous system during these early stages was determined by immunocytochemistry using a polyclonal antibody. Whole mounts of chick embryos at 24-30-h stages showed intense immunoreactivity of the neural tube in formation along its full extent. Cross-sections at these stages of development showed strong labeling in neuroepithelial cells at the basal and apical regions of the neural tube wall. Embryos at more advanced periods of development (48h and 72 h) showed distinctive immunolabeling of neuroepithelial cells, neuroblasts and their cytoplasmic extensions in the mantle layer of the stratified neural tube wall, and neuroblasts and their cytoplasmic extensions in the internal wall of the optic cup, as well as a striking labeling of cells in the apparent nuclei of cranial nerves and budding fibers. These immunolocalization studies indicate temporal and site-specific expression of myosin Va during chick embryo development, suggesting that myosin Va expression is related to recruitment for specific cellular tasks.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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This study was carried out to assess the influence of bovine embryo culture medium Beltsville Agriculture Research Center (BARC), supplemented with FCS, BSA or PVA, on the in vitro oocyte maturation, evidenced by cleavage rate and blastocysts production at different developmental stages. Three experiments were performed, as follows: exp.1: addition of FCS to BARC medium at concentrations of 0, 5 and 10%; exp. 2: addition of BSA to BARC medium at concentrations of 0, 4 and 8 mg/ml; exp. 3: addition of PVA to BARC medium at concentrations of 0, 0.5 and 1.0 mg/ml. TCM 199 supplemented with bicarbonate, pyruvate, gentamicin sulfate, FSH, LH and FCS was used as control group. Oocytes obtained from cow ovaries at slaughterhouse were selected in PBS, and then matured in BARC medium supplemented with FSH, LH and gentamicin sulfate, according to the experimental design. Percoll gradient was used for sperm selection and TALP medium for IVF. In vitro embryo culture was in SOF-m medium; a humidified atmosphere with 5% CO2, in air, at 38.7oC was used for all steps. The number of oocytes reaching blastocyst, expanded blastocyst, and hatched blastocyt stages was recorded, respectively at 72 and 168 h post-insemination. ANOVA and Bonferroni t test were used to determine differences among groups. Differences of P<0.05 were taken as significant. Higher percentage (P<0.05) of cleaved oocytes was observed in group TCM + FCS than for the other groups matured in BARC supplemented with FCS or BSA, regardless the concentration used. However, the cleavage rate was similar between groups BARC plus PVA with 1 mg/ml (85.7%) and TCM + FCS (90.8%). Significant difference was found among groups for the production of blastocysts, with the control group yielding a higher number of blastocysts (results ranging from 47.4 to 51.4%, in comparison with groups using BARC + FCS (4.1 to 19.7%), BSA (1.4 to 5.6%) and PVA (5.7 to 10.6%). In conclusion, BARC medium supplemented with different macromolecules did not promote a beneficial effect on in vitro oocyte maturation, resulting in lower rate of cleavage and blastocyst production when compared with TCM + FCS medium.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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This study tested the effect of Sigma antioxidant supplement®, α-tocopherol (vitamin E) and L-ascorbic acid (vitamin C) in the culture medium of bovine embryos. In experiment 1, in vitro produced bovine zygotes were cultured in Human Tubal Fluid (HTF): Eagle’s Basic Medium (BME) with: Group 1 – 50 µm vitamin C; Group 2 – 200 µm vitamin E; Group 3 – 25 µm vitamin C and 100 µm vitamin E; Group 4 – 1 µl/ml Sigma antioxidant supplement®; and the Control group – HTF:BME only. In experiment 2, embryos were cultured in high or low oxygen tension with HTF:BME + Sigma antioxidant supplement® or in HTF:BME alone (Control). The data were analyzed using ANOVA followed by Tukey’s test. The results of experiment 1 showed a negative effect (P < 0.05) of vitamin E on blastocyst production in Group 2 (19.7 ± 0.1%). This effect was reduced in Group 3 by the addition of vitamin C (26.1 ± 0.2%). The use of vitamin C alone (34.9 ± 0.3%) or the Sigma antioxidant supplement® (33.3 ± 0.7%) did not increase (P > 0.05) the number of blastocysts produced compared with the control group (30.1 ± 0.5%). During experiment 2, there was no effect (P > 0.05) from the culture medium or the O2 concentrations used, indicating that the reduction of the O2 concentration did not improve blastocyst production.
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Fundação de Apoio à Pesquisa do Estado de São Paulo (FAPESP)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)