female cd1 mice  (Charles River Laboratories)

Journal: Development (Cambridge, England)

Article Title: Dynamic WT1 expression during gastrulation specifies peritoneal smooth muscle fate independently of mesothelial fate

doi: 10.1242/dev.204332

Figure Lengend Snippet: Embryonic contribution of cells of the Wt1 lineage to visceral mesothelium or visceral and vascular smooth muscle is dependent on timing of tamoxifen dosing. (A) Schematic of the study design: CD1 dams were time-mated with Wt1 CreERT2/+ ; Rosa26 lacZ/ l acZ males and received tamoxifen (Tam) at the respective embryonic stages (embryonic day, E); embryos were dissected between E17.5 and E19.5, and XGal stained. (B) When Tam was given at E13.5 and embryos analysed at E17.5, the mesentery (M) and intestine (I) were well covered by XGal + ( lacZ -expressing) cells. (C,D) When Tam had been given at E12.5, the coverage of XGal-positive cells in the mesentery was less complete at E19.5. This loss of coverage of XGal-positive cells was still more pronounced E17.5 when Tam had been given at E11.5 (D). (E) Similarly, there were only patches of XGal-positive cells in the mesentery and over the intestine at E17.5 after Tam administration at E9.5. (F,G) When Tam had been given at E8.5, XGal-positive cells were observed in small patches in the mesentery (arrow, F), or in cell chains near the vasculature (arrowhead, F), or in distinct rings (unfilled arrowheads, G) surrounding the mesenteric veins (filled arrowhead, G) in the second arcade of the mesenteric vascular tree. The corresponding artery is depicted with an arrow (G). (H-J) Tam dosing at E7.5 resulted at E18.5 in XGal-positive cells scattered within the mesentery (H), in the circular musculature of the small intestine (white arrowheads, I) and surrounding a segment of mesenteric artery of the second arcade (black arrowheads, J). Scale bars: 500 μm (B-D,F,H); 200 μm (E,G,I,J).

Article Snippet: CD1 female mice (at least 10 weeks old, Charles River, Harlow, UK) were time-mated with Wt1 CreERT2/+ ; Rosa26 lacZ/lacZ or Wt1 CreERT2/+ ; Rosa26 mTmG/+ males, and noon of the day on which a vaginal plug was detected was considered as E0.5.

Techniques: Staining, Expressing

Journal: Development (Cambridge, England)

Article Title: Dynamic WT1 expression during gastrulation specifies peritoneal smooth muscle fate independently of mesothelial fate

doi: 10.1242/dev.204332

Figure Lengend Snippet: Contribution of Wt1 lineage cells to the mesentery near developing vasculature and to the intestinal wall. (A) Schematic of the study design: embryos were analysed at E12.5 or E15.5 after timed mating of Wt1 CreERT2/+ ; Rosa26 mTmG/mTmG males with CD1 females and Tam dosing at E8.5. (B,B′) View of the intestinal loop and mesentery. GFP-positive cells (arrowheads) were found within the mesentery of the developing intestinal tract (arrows; asterisks mark the caecum). Note the abundant GFP fluorescence in the liver (Li). (C,D) Immunofluorescence of CD31 and GFP on sections through this region showed GFP-positive cells within the mesentery and in the intestinal wall, near the developing microvascular plexus (C). Higher magnification (D) reveals that GFP-positive cells (arrowheads) are often arranged close to neighbouring endothelial cells (arrows) of the vascular plexus in the intestinal wall and mesentery. (E-F″) Immunofluorescence for PDGFRB and GFP on neighbouring sections showed that GFP-positive cells within the intestinal wall mesenchyme and mesentery co-expressed PDGFRB (arrowheads), while a few mesothelial GFP-positive cells were PDGFRB negative (F-F″, arrows). (G,H) Representative images from two different embryos at E15.5 after Tam dosing at E8.5. GFP-positive cells (arrowheads) were found within a segment of the small intestine in a circular arrangement. e, endoderm; m, mesentery; ma, mesenteric artery; mv, mesenteric vein. Scale bars: 500 μm (G,H); 300 μm (B,B′); 100 μm (C,E,F-F″); 50 μm (D).

Article Snippet: CD1 female mice (at least 10 weeks old, Charles River, Harlow, UK) were time-mated with Wt1 CreERT2/+ ; Rosa26 lacZ/lacZ or Wt1 CreERT2/+ ; Rosa26 mTmG/+ males, and noon of the day on which a vaginal plug was detected was considered as E0.5.

Techniques: Fluorescence, Immunofluorescence