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Fig. 6 | BMC Developmental Biology

Fig. 6

From: Analysing bioelectrical phenomena in the Drosophila ovary with genetic tools: tissue-specific expression of sensors for membrane potential and intracellular pH, and RNAi-knockdown of mechanisms involved in ion exchange

Fig. 6

RNAi-knockdown of ork1 in the FE results in spherical follicles with altered cytoskeletal organisation. a-d Brightfield-images of follicles from tj-Gal4 > ork1 shRNA (a-c) and mat-tub-Gal4-GeneSwitch > ork1 shRNA ovaries (control; d). a For soma knockdown of ork1, the tj-Gal4 driver was used. Ovaries of all analysed flies contained spherical follicles and eggs (S8-S14; scale bar represents 100 μm). b and c Brightfield-images of S10B- and S12-tj-Gal4 > UAS-ork1 shRNA follicles; the oocyte nucleus (dorsal) is marked with an asterisk (scale bars represent 50 μm). d Ovaries from mat-tub-Gal4-GeneSwitch > UAS-shRNA flies, having a low transcription level of ork1-shRNA, were used as control (scale bar represents 100 μm). Similar to ovaries from strong germline knockdowns via the mat-tub-Gal4 and MTD-Gal4 drivers (cf. Table 1), ovaries from control flies only produced follicles resembling the wt. e In contrast to wt, ork1-follicles (S9 and S10B) exhibit a weaker microtubule (MT) cytoskeleton in the FE, and the MT are not aligned along the a-p axis (scale bars represent 10 μm). Tangential optical sections (SIM) of typical anti-acetylated α-tubulin-treated follicles are shown. f Concerning basal microfilaments (bMF) in the FE, ork1 exhibits even stronger anomalies in comparison to wt. Tangential optical sections of typical S9, S10A, S10B and S12 wt- and ork1-follicles stained with fluorescent phalloidin are shown (cFC, centripetal FC; mFC, mainbody FC; vFC, ventral FC; a, anterior; p, posterior; d, dorsal; v, ventral); d-v orientation, as indicated, applies to all images, except vFC-images (scale bars represent 10 μm). Since, due to bMF-condensations in wt-follicles, it is difficult to reveal transversal bMF-alignment in dorsal and lateral cFC [5, 17], vFC are shown for S10B. Typical bMF-condensations (asterisks), as in S9 cFC in wt, are missing in ork1. The bMF in ork1 S10A show the same parallel alignment as in wt, however, in some areas (arrowheads), the bMF-cytoskeleton is weaker. During S10B and S12, ork1-follicles are characterised by a disturbed transversal transcellular bMF-alignment, resembling the kugelei mutant [22]

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