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Comparison of proteins synthesized by polarized caprine oviductal epithelial cells and oviductal explants in vitro

机译:极化的山羊输卵管上皮细胞和输卵管外植体合成蛋白的体外比较

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Our objectives were to compare proteins secreted by caprine oviductal explants and oviductal epithelial (OE) cells in vitro. Oviducts were collected from goats on Days 1 (n = 5) and 5 (n = 5) of the estrous cycle. Radiolabeled secretory proteins from tissue segments and cell cultures were visualized using SDS-PAGE and fluorography. After culture, media from ampulla oviduct segments collected on Days 1 and 5 of the estrous cycle contained an acidic 97 kDa protein, which was greatly reduced in culture medium obtained from infundibulum and isthmus oviduct segments. A complex of low molecular weight proteins (14-26 kDa) could be modulated by estradiol when OE cells were cultured on plastic. This complex was constitutively expressed when OE cells were cultured on Matrigel-coated filters. Polarized OE cells were also capable of compartment-specific secretion of [(L)-S-35]-methionine-labeled proteins. A 45 kDa acidic protein was predominantly secreted into the apical compartment while a 66 kDa acidic protein was preferentially localized in the basal compartment. Proteins secreted by OE cells were similar to proteins secreted by tissue segments in vitro. Therefore, under well-defined culture conditions OE cells may be useful in enhancing in vitro fertilization or early embryonic development.
机译:我们的目标是在体外比较山羊输卵管外植体和输卵管上皮(OE)细胞分泌的蛋白质。在发情周期的第1天(n = 5)和第5天(n = 5)从山羊收集输卵管。使用SDS-PAGE和X射线照相术可观察到来自组织片段和细胞培养物的放射性标记分泌蛋白。培养后,在动情周期第1天和第5天从壶腹输卵管段收集的培养基中含有一种酸性97 kDa蛋白,在从漏斗和地峡输卵管段获得的培养基中该蛋白大大减少。当在塑料上培养OE细胞时,雌二醇可调节低分子量蛋白质(14-26 kDa)的复合物。当在基质胶包被的滤膜上培养OE细胞时,此复合物组成型表达。极化的OE细胞也能够[[(L)-S-35]-蛋氨酸标记的蛋白质的隔室特异性分泌。 45 kDa的酸性蛋白主要分泌到根部腔室中,而66 kDa的酸性蛋白则优先定位在基底腔室中。 OE细胞分泌的蛋白质与体外组织片段分泌的蛋白质相似。因此,在条件明确的培养条件下,OE细胞可用于增强体外受精或早期胚胎发育。

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