Reversible Interconversion between Primitive Endoderm- and Parietal Endoderm-Like F9 Cells Demonstrated by mRNAs Expression

Abstract

The differentiation of retinoic acid-treated F9 cells (primitive endoderm-like F9 cells) into parietal endoderm-like F9 cells induced by dibutyryl cAMP was studied as a culture model of the morphogenesis of early mouse embryo. For this purpose, 6 cDNA clones coding for mRNAs specifically expressed in parietal endoderm-like F9 cells were selected. Northern hybridization of RNA extracted from variously treated F9 cells to nick-translated plasmid DNA of these clones demonstratedthe reversible expression of many mRNAs depending on the presence of dibutyryl cAMP in the culture medium. This result suggested that the differentiated state of parietal endoderm, which is formed from primitive endoderm at a position ad-jacent to the trophectoderm in mouse embryo, can be reversed if the local signal isremoved. One of the selected clones, pLAM, hybridized to an mRNA of 6.3kb andselected mRNA producing a laminin B subunit in an in vitro translation system. This clone has an inserted sequence of 3.1kb. Among the restriction sites in this sequence, six were consistent with those in a 1.7kb inserted sequence of pPE 49 and pPE 386, which were isolated by Barlow et al. as laminin B₁ clones. An XbaI site found in both pPE 49 and pPE 386 was, however, not found at the corresponding position of pLAM. Dot hybridization of RNA with pLAM showed that expression of laminin B in F9 cells is stimulated more than 100-fold during differentiation of F9 stem cells into parietal endoderm-like F9 cells.