Detection of HEX 70a in ovarioles of workers at the beginning of the pharate-adult development (∼1 day after pupal ecdysis) (the developmental stage is illustrated at the upper left corner of the figure).

Abstract

<p>(<b>A</b>) Light microscopy of ovarioles (covered by their respective peritoneal sheath) stained with methylene blue/basic fuchsin. Only the germarium is focused in this figure (the most anterior region of the ovariole, or terminal filament, is not shown). A rosette formed by germline cells (oocyte and nurse cell precursors) is distinguishable (circle) in the germarium. (<b>B, C</b>) Confocal microscopy image of rhodamine/phalloidin labeled F-actin (green) and DAPI-labeled cell nuclei (blue) showing aspects of the structure of the ovarioles (peritoneal sheath removed) at the time they were used for HEX 70A detection. The actin-rich polyfusomes (arrowheads in B) are seen in the center of the cystocyte rosettes in the upper region of the germarium. Ring canals derived from polyfusomes (arrows in B and C) are apparent in the lower region of the germarium shown in B and in higher magnification in C. (<b>D</b>) Confocal microscopy of an ovariole (upper portion of the germarium) stained with DAPI. (<b>E</b>) The same ovariole showing foci of HEX 70a detected with anti-HEX 70a/Cy3 (red). (<b>F</b>) The merged D and E images. The insert in F shows a “control” ovariole (upper portion of the germarium) incubated with the pre-immune serum and subsequently stained with Cy3/DAPI. Arrowheads in D-F show nuclei of germline cells. Arrows in D–F point to nuclei of follicle cell precursors. In all figures, the upper portion of the germarium is oriented upward.</p