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Fig. 2. Localization of Dgo during PCP establishment in the eye. Anterior is
leftwards and dorsal upwards in all panels except F and I, where proximal is
leftwards. (A) Single apical confocal microscopy section showing ommatidial
rows 1-7 stained with anti-Dgo (green) and phalloidin (highlighting F-actin,
red) to mark the center of each cluster. Morphogenetic furrow (anterior) is on
the left of the panel and the equator is at the bottom. (Right panels)
Monochrome higher magnifications of single clusters from row 3, row 5 and row
6 (as indicated). Dgo is first detected apical at the membrane in all cells
ahead of and posterior to MF (up to row 2). More posteriorly, the initial
uniform apical localization changes (row 3) within five cell preclusters, and
Dgo is present at higher levels in R3/4 cells within the equatorial-polar (DV)
axis. (Row 5) Dgo is maintained at the border between the R3/R4 cells, but it
is now detected less in parts of R3 and at higher levels in R4. (Row 6) The R4
specific enrichment is now complete and persists through row 10. Whereas there
is no Dgo detected at membranes where R3/R4 abut the R2/R5 pair, Dgo is
enriched at the opposing R8, R2/R5 membranes. (B-D) Mosaic analysis of GFP-Dgo
to determine the precise localization of Dgo at the R3/R4 cell boundary.
GFP-Dgo is in green, anti-DE Cadherin is in red. Right side panels show a
schematic of actual clusters on the left. Cells are numbered according to
their position; cells shown in white express GFP-Dgo in the given cluster;
GFP-Dgo at localization membranes is highlighted with green lines. (B,C)
Mosaic clusters in row 3 with either R4 (B) or R3 (C) expressing GFP-Dgo. Dgo
localization reflects a horseshoe-like pattern in both R3 and R4 cells. (D,E)
Mosaic clusters in rows 5-6, with (again) either R4 (D) or R3 (E) expressing
GFP-Dgo. Dgo is now enriched only at the R3 side of the R3/R4 cell boundary
and the polar side of R4 (very similar to the localization of Fz). (F) GFP-Dgo
(green) clones in pupal wings at 60-80 hours at 18°C. lacZ (blue)
marks the cells that express GFP-Dgo; F-actin (phalloidin, red) labels the
growing actin hairs; proximal is leftwards. GFP-Dgo localizes to the distal
membrane of each cell that expresses it. (G-I) Schematic summary presentation
of the Dgo localization patterns in developing eye (G, row 3; H, row 6) and
pupal wing (I) cells; again, Dgo shows the same localization pattern as Fz and
Dsh.
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