Figure S4.
Ciliary structures are disrupted in haltere receptors, but not in leg receptors. (A) Localizations of Ana1-tdTom, Mks1-GFP, and B9d1-RFP (Basiri et al., 2014) in haltere pedicellar campaniform mechanoreceptors in wild type (upper) and c01236/BE6 (lower). Ana1-tdTom (centriole marker) in c01236/BE6 showed a comparable pattern with wild-type cells, consistent with the presence of two centrioles observed in our ET data. Mks1-GFP (transition zone marker) was present in nearly all mutant receptors, but some of them showed a half-open ring shape, consistent with the enlargement of the cilium in c01236/BE6. B9d1-RFP (transition zone marker) was nearly absent in all mutant receptors. These observations confirmed that the ciliary structure was disrupted in the haltere receptors in c01236/BE6. Upper left: Ana1-tdTom. Upper middle: Mks1-gfp. Upper right: B9d1-rfp. Lower left: Ana1-tdTom; c01236/BE6. Lower middle: Mks1-gfp; c01236/BE6. Lower right: B9d1-rfp; c01236/BE6. Scale bar, 5 µm. (B) Localizations of Ana1-tdTom, Mks1-GFP, and B9d1-RFP in leg receptors in wild type (upper) and c01236/BE6 (lower; n = 10 for each genotype). In mutant flies, the external cupola structure (white arrowhead) was used to locate the signal of ciliary markers. The centriole (Ana1-tdTom) and transitional zone (Mks1-GFP and B9d1-RFP) markers appeared to be normal, suggesting that the ciliary structures are intact in mutant leg receptors. Upper left: dcx-emap-gal4/Ana1-tdTom; +/uas-cd4-tdgfp. Upper middle: dcx-emap-gal4/Mks1-gfp; +/uas-cd4-tdTom. Upper right: dcx-emap-gal4/B9d1-rfp; +/uas-cd4-tdgfp. Lower left: dcx-emap-gal4/Ana1-tdTom; c01236/BE6. Lower middle: dcx-emap-gal4/Mks1-gfp; c01236/BE6. Lower right: dcx-emap-gal4/B9d1-rfp; c01236/BE6. Scale bar, 5 µm. (C and D) Representative ET slice images of the lateral view of the sensory cilium in a mutant fly (c01236/BE6). Note that the ciliary structures (i.e., centriole and cilium) were intact. The region in the dashed box in C was enlarged in D. Scale bars, 500 nm. OS, outer segment. (E) Statistical quantification of dendritic length of leg receptors (from confocal images) in wild type (n = 4 cells) and c01236/BE6 (n = 4 cells). (F) Statistical quantification of the cross-section area of dendrite in haltere receptors (from FIB/scanning electron microscopy volume data) in wild type (n = 4 cells) and c01236/BE6 (n = 4 cells). In E and F, data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. n.s., no significance; mem, membrane.

Ciliary structures are disrupted in haltere receptors, but not in leg receptors. (A) Localizations of Ana1-tdTom, Mks1-GFP, and B9d1-RFP (Basiri et al., 2014) in haltere pedicellar campaniform mechanoreceptors in wild type (upper) and c01236/BE6 (lower). Ana1-tdTom (centriole marker) in c01236/BE6 showed a comparable pattern with wild-type cells, consistent with the presence of two centrioles observed in our ET data. Mks1-GFP (transition zone marker) was present in nearly all mutant receptors, but some of them showed a half-open ring shape, consistent with the enlargement of the cilium in c01236/BE6. B9d1-RFP (transition zone marker) was nearly absent in all mutant receptors. These observations confirmed that the ciliary structure was disrupted in the haltere receptors in c01236/BE6. Upper left: Ana1-tdTom. Upper middle: Mks1-gfp. Upper right: B9d1-rfp. Lower left: Ana1-tdTom; c01236/BE6. Lower middle: Mks1-gfp; c01236/BE6. Lower right: B9d1-rfp; c01236/BE6. Scale bar, 5 µm. (B) Localizations of Ana1-tdTom, Mks1-GFP, and B9d1-RFP in leg receptors in wild type (upper) and c01236/BE6 (lower; n = 10 for each genotype). In mutant flies, the external cupola structure (white arrowhead) was used to locate the signal of ciliary markers. The centriole (Ana1-tdTom) and transitional zone (Mks1-GFP and B9d1-RFP) markers appeared to be normal, suggesting that the ciliary structures are intact in mutant leg receptors. Upper left: dcx-emap-gal4/Ana1-tdTom; +/uas-cd4-tdgfp. Upper middle: dcx-emap-gal4/Mks1-gfp; +/uas-cd4-tdTom. Upper right: dcx-emap-gal4/B9d1-rfp; +/uas-cd4-tdgfp. Lower left: dcx-emap-gal4/Ana1-tdTom; c01236/BE6. Lower middle: dcx-emap-gal4/Mks1-gfp; c01236/BE6. Lower right: dcx-emap-gal4/B9d1-rfp; c01236/BE6. Scale bar, 5 µm. (C and D) Representative ET slice images of the lateral view of the sensory cilium in a mutant fly (c01236/BE6). Note that the ciliary structures (i.e., centriole and cilium) were intact. The region in the dashed box in C was enlarged in D. Scale bars, 500 nm. OS, outer segment. (E) Statistical quantification of dendritic length of leg receptors (from confocal images) in wild type (n = 4 cells) and c01236/BE6 (n = 4 cells). (F) Statistical quantification of the cross-section area of dendrite in haltere receptors (from FIB/scanning electron microscopy volume data) in wild type (n = 4 cells) and c01236/BE6 (n = 4 cells). In E and F, data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. n.s., no significance; mem, membrane.

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