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Fig. 5. Extruded LECs are removed by circulating haemocytes. (A)
Macrophage-like haemocytes circulate under the pupal epidermis basal lamina
extending and retracting leading lamellipodia (arrowhead). Haemocytes were
visualized in vivo through the pupal cuticle by the expression of
membrane-bound Src-GFP (green) using a haemocyte-specific driver (Srp-Gal4).
(B) Travelling cells in the haemolymph are recruited to the basal
surface of LECs. The actomyosin cytoskeleton of LECs was visualized by the
expression of Sqh-GFP using confocal XZT acquisition. The apical constriction
of LECs (small arrowheads) was followed by the attachment of bright
fluorescent bodies to their basal side (large arrowhead). (C) The
recruitment of haemocytes to the basolateral surface of LECs occurs
sequentially to constriction. Simultaneous in vivo visualization of LECs and
histoblasts (ubiquitous expression of Stb-YFP, red) and haemocytes
(Srp-Gal4/UAS-GFP, green) (see Movie 10 in the supplementary material).
Snapshots show LECs undergoing apical constriction (arrowhead) and being
engulfed from their basal surface by haemocytes (asterisk) extending
cytoplasmic projections (small arrowheads). Transverse z-projections
show that encapsulation initiates before LEC delamination is completed and is
extremely fast. The whole process (apical constriction, delamination and
engulfment) takes place in approximately 45 minutes.
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