PROGRESSIVE EXPRESSION OF TROPHOBLAST-SPECIFIC GENES DURING FORMATION OF MOUSE TROPHOBLAST GIANT-CELLS INVITRO
PROGRESSIVE EXPRESSION OF TROPHOBLAST-SPECIFIC GENES DURING FORMATION OF MOUSE TROPHOBLAST GIANT-CELLS INVITRO
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DOI：
10.1002/mrd.1080340403
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发表时间：
1993-04-01
影响因子：
2.5
通讯作者：
ROSSANT, J
中科院分区：
文献类型：
--
作者：
CARNEY, EW;PRIDEAUX, V;ROSSANT, J
在体内和体外滋养外胚层发育过程中研究了一系列滋养层特异性 mRNA 的表达，以评估这些 mRNA 作为滋养层分化标记的用途，并检查各种滋养外胚层衍生物之间的谱系关系。对切片的第 6.5-18.5 天小鼠胚胎进行原位杂交，将小鼠胎盘催乳素 I 和 II 以及小鼠增殖素 (PLF) 的 mRNA 定位到滋养层巨细胞，并将增殖素相关蛋白 mRNA 定位到海绵滋养层和巨细胞层。第五个标记 cDNA 4311 仅在海绵滋养层中发现。然后培养第3.5天的囊胚生长物和第7.5天的二倍体胚外外胚层(EX)和外胎盘锥(EPC)以在体外培养多倍体巨细胞。 2、4或6天后对培养物进行原位杂交处理。 EX和EPC均形成次级巨细胞，其以与体内观察到的相同序列表达所有标记物，并且囊胚生长物中的初级巨细胞在培养的第4天和第6天表达早期巨细胞标记物PLF和PL-1。 EPC 比 EX 早 2 天完成序列，表明 EPC 致力于巨细胞形成。这些结果表明，EX、EPC以及初级和次级巨细胞都共享共同的分化途径，并且该途径的基因表达特征的高度有序序列在体内和体外发生相似。
The expression of a battery of trophoblast-specific mRNAs was studied during trophectoderm development in vivo and in vitro to assess the use of these mRNAs as markers of trophoblast differentiation and to examine lineage relationships between various trop ectoderm derivatives. In situ hybridization of sectioned day 6.5-18.5 mouse embryos localized mRNAs for mouse placental lactogens I and II and mouse proliferin (PLF) to trophoblast giant cells and proliferin-related protein mRNA to the spongiotrophoblast and giant cell layers. A fifth marker, cDNA 4311, was found only in spongiotrophoblast. Day 3.5 blastocyst outgrowths and day 7.5 diploid extraembryonic ectoderm (EX) and ectoplacental cone (EPC) were then cultured to produce polyploid giant cells in vitro. Cultures were processed for in situ hybridization after 2, 4, or 6 days. EX and EPC both formed secondary giant cells, which expressed all markers in the same sequence as was observed in vivo, and primary giant cells in blastocyst outgrowths expressed the early giant cell markers PLF and PL-1 on days 4 and 6 of culture. EPC progressed through the sequence 2 days ahead of EX, indicating commitment of EPC to giant cell formation. These results suggest that EX, EPC, and primary and secondary giant cells all share in a common pathway of differentiation and that the highly ordered sequence of gene expression characteristic of this pathway occurs similarly in vivo and in vitro.