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Fig. 3. Early patterning of the mouse chorion. (A) In situ hybridizations for Hand1 and Pl1 at E9.0 indicate that Hand1⁺/Pl1^- cells line the maternal blood sinuses located on the apical side of the chorion. At E8.5, Syna expression is restricted to cells near the top of the chorion, whereas Gcm1, Synb and Cebpa staining (on serial sections) indicates co-localization within a subset of trophoblast located on the basal side of the chorion (confirmed by double in situ hybridization, data not shown). The black arrows indicate Hand1⁺/Pl1^- cells lining maternal sinusoids. (B) Higher magnification images of Hand1 and Syna in situ hybridizations on E8.5 chorion. Note that Syna-positive cells are separated from the maternal blood sinuses by a single layer of cells, typically Hand1-positive. Black arrows indicate the cell layer lining the maternal sinusoids. (C) Ultrathin epoxy section of E8.5 chorion (2 µm) stained with Tissue Epoxy Stain (EMS). Note the unique morphology of the different layers of trophoblast cells in the chorion. Cells that line the maternal sinusoids (Hand1⁺/Pl1^- at E8.5) are positive for Ctsq expression by E12.5 (data not shown). Scale bars: black, 100 µm; red, 50 µm.

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