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Figure 5


Fig. 5. Production of hindbrain choroid plexus epithelium begins at ~E9.5, peaks between ~E11 and E12, and terminates by ~E14. (A-F) Transverse sections through E9.5-E14.5 hindbrain that has been processed to detect FlpeERT2 mRNA show expression limited to the rhombic lip (vertical brackets in A-C), and absent from the hindbrain roof plate epithelium (hRPe, opposing black arrowheads) and hindbrain choroid plexus epithelium (hCPe). (G-L) Coronal sections processed for PLAP detection (dark precipitate) after harvest from E16.5 doubly transgenic Wnt1::FlpeERT2; R26::FRAP embryos (n=9). Induction of FlpeERT2-mediated recombination by tamoxifen (TAM) administration separates temporal cohorts of Wnt1 (rhombic lip)-derived hCPe. (G) E9.5 TAM administration (recombination at ~E10 in the rhombic lip) resulted in a few labeled E16.5 hCPe cells. (H-J) TAM at E10.5 (recombination at ~E11), E11.5 (recombination at ~E12) or E12.5 (recombination at ~E13) resulted in numerous labeled cells in the E16.5 hCPe. (K,L) TAM administration at E13.5 (recombination at ~E14) resulted in few labeled hCPe cells (K), and none were present after administration at E14.5 (L). Insets in G-L reveal no PLAP activity when diluent alone was administered. (M-R) Coronal sections from doubly transgenic Wnt1::cre; Cre-responsive ßgal indicator embryos, with ßgal expression (green by immunodetection) distinguishing hCPe from underlying mesenchyme and vasculature, and red indicating BrdU incorporation, reflecting birthdate (time of single BrdU injection indicated above each panel). BrdU administered at either E9.5 or E14.5 showed very few co-labeled hCPe cells at E16.5 (M,R, respectively), whereas BrdU administered from E10.5 to E13.5 showed numerous co-labeled hCPe cells (N-Q). Insets show higher-power views of the boxed areas. rec, recombination time point; harv, harvest time point; hCP, hindbrain choroid plexus; LRL, lower rhombic lip.





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