Fig. 4

Screening strategy for neurodevelopmental defects in the developing fly eye. a Schematics and images of the wild-type adult, pupal, and larval eye show the cell organization and structure of the fly eye during development. The wild-type adult eye displays a symmetrical organization of ommatidia, and Flynotyper software detects the center of each ommatidium (orange circle) and calculates a phenotypic score based on the length and angle between the ommatidial centers. Illustrations of the wild-type pupal eye show the arrangement of cone cells (C), primary pigment cells (1°), and secondary pigment cells (2°) along the faces of the hexagon, and bristle cells (b) and tertiary pigment cells (3°) at alternating vertices, as well as the eight photoreceptor cells within an ommatidium. The larval imaginal disc schematic shows proliferating cells posterior to the morphogenetic furrow. Pupal eyes were stained with anti-Dlg and phalloidin to visualize ommatidial cells and photoreceptor cells, respectively, while the larval eye was stained with anti-pH3 to visualize proliferating cells. Diagrams of the pupal and larval eye were generated by Frank Pichaud⁶⁹ and Joan E. Hooper⁷⁰ and are reprinted with permission from the publishers. b Example images of pupal eyes stained with anti-Dlg illustrate the structure and organization in control and knockdown flies. Circles and arrows indicate differences in cell organization between control and knockdown pupal eyes (yellow circles: cone cell number and organization, white circles: bristle groups, white arrowheads: secondary cells, white arrows: primary cells, yellow arrows: rotation of ommatidia)