Analysis of proteins secreted by mouse embryos developing in vivo and in vitro.
Analysis of proteins secreted by mouse embryos developing in vivo and in vitro.
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体内和体外发育的小鼠胚胎分泌的蛋白质分析。
DOI：
10.1002/jez.1402520205
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Nieder,GL
中科院分区：
文献类型：
--
作者：
Nieder,GL
将体外发育的小鼠胚泡分泌的蛋白质与子宫内发育的胚泡分泌的蛋白质进行比较，以确定支持胚泡发育的简单培养基是否也支持分泌的蛋白质表达。在妊娠第3、4或5天收集体内胚胎，并在35 S-蛋氨酸中孵育，以产生含有释放的标记蛋白的条件培养基。在第3天收集用于培养的胚胎，并在48或72小时后产生标记的条件培养基。通过二维电泳分离标记的蛋白质，并使用数字图像分析系统进行比较。第3天胚胎没有释放可检测量的蛋白质，尽管细胞内蛋白质的合成是大量的。第4天和第5天囊胚释放的蛋白质数量和复杂性增加，与先前的结果一致。当第3天的胚胎在含有4 mg/ml BSA的培养基中培养48 h时，分泌的蛋白质模式与第5天的子宫囊胚相似但不相同。虽然子宫胚泡产生的大多数蛋白质是由培养的胚胎分泌的，但某些蛋白质的相对量存在差异。4 mg/ml的结晶BSA和聚乙烯醇均不支持蛋白分泌以及粗级分-V BSA的形成。局限于输卵管的囊胚与子宫囊胚相比，蛋白质分泌模式也表现出数量上的差异。因此，尽管囊胚发育和许多分泌蛋白的表达在子宫外得到支持，但着床胚胎的分泌特征的完整模式可能取决于特定的子宫影响。
Proteins secreted by mouse blastocysts developing in vitro were compared to those from blastocysts developing in utero to determine if a simple medium supporting blastocyst development also supports secreted protein expression. In‐vivo embryos were collected on days 3, 4, or 5 of pregnancy and incubated in35S‐methionine to produce conditioned medium containing released, labeled proteins. Embryos for culture were collected on day 3 and after 48 or 72 h labeled conditioned medium was produced. Labeled proteins were separated by two‐dimensional electrophoresis and compared using a digital image analysis system. Day 3 embryos did not release proteins in detectable amounts, although synthesis of intracellular proteins was substantial. Day‐4 and ‐5 blastocysts released proteins in increasing amount and complexity, consistent with previous results. When day‐3 embryos were cultured in medium containing 4 mg/ml BSA for 48 h, secreted protein patterns were similar but not identical to those of day‐5 uterine blastocysts. Although most of the proteins produced by uterine blastocysts were secreted by cultured embryos, differences were found in the relative quantities of certain proteins. Neither crystallized BSA nor polyvinyl alcohol at 4 mg/ml supported development of protein secretion as well as the crude fraction‐V BSA. Blastocysts restricted to the oviduct also exhibited quantitative differences in protein secretion patterns compared to uterine blastocysts. Thus, although blastocyst development and the expression of many secreted proteins are supported outside the uterus, the full pattern of secretion characteristic of the peri‐implantation embryo may be dependent on specific uterine influences.