Figure 3.
Polarity and stability of microtubules in the outer segment of fly campaniform receptors. (A) Localization of Patronin-RFP (patronin-rfp-KI) in haltere receptors (lateral view). White arrowhead, Patronin-RFP in the TB. Inset (top view), distal (upper) and proximal (lower) ends of the MO (patronin-rfp-KI; dcx-emap-gal4/uas-cd4-tdgfp). Yellow arrowhead, two dot-shaped Patronin-RFP signals outside the neuron (Fig. S1). Scale bar, 1 µm. (B) Localization of Patronin-RFP in leg receptors (lateral view). White arrowhead, the distal signal of Patronin-RFP (inset, 600 nm from the distal tip). Blue arrowhead, the proximal signal of Patronin-RFP. Yellow arrowheads, two dot-shaped Patronin-RFP signals outside the neuron (Fig. S1). Scale bar, 1 µm. (C) EB1 comets (EB1-tdgfp-KI) in leg receptors. Left panel: A snapshot of EB1 comets (Video 2). White arrow, MO. Yellow arrowheads, two dot signals associated with the microtubules outside the neuron. Right panel: The trajectories of EB1 comets in 3 min. White arrowhead: dendritic inner segment (d). Scale bars, 5 µm. (D) Intensity line profile of EB1 signal in the outer segment along the proximal–distal axis (inset). The signals at the distal end (Id) and at the proximal part (Ip) were quantified. Scale bar (inset), 3 µm. (E) Statistical quantification of EB1 signal at the distal end (Id) and the proximal end (Ip; n = 5 cells). The data were presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. ***, P < 0.001. (F) Three sets of representative confocal images (−1 s: right before bleaching; 0 s: just after bleaching; 3 h or 9 min: time after bleaching) showing the recovery of GFP-tubulin (dcx-emap-gal4/uas-gfp-αtub84B) or EB1-GFP (dcx-emap-gal4; uas-EB1-gfp). Scale bars, 5 µm. (G) Fluorescence recovery curves after photobleaching. Red (circle), GFP-tubulin recovery in the outer segment (n = 10 cells). Black (square), GFP-tubulin recovery in the inner segment (n = 9 cells). Blue (triangle), EB1-GFP recovery in the outer segment (n = 6 cells). Data were presented as mean ± SEM. (H) Cartoon schematic of four groups of microtubules. 1, microtubules in the MO. 2, microtubules in the TB. 3, microtubules in the inner segment. 4, microtubules in the thecogen cell. +, plus end; −, minus end; BB, basal body; EDM, electron-dense materials; IS, inner segment; OS, outer segment; mem, membrane.

Polarity and stability of microtubules in the outer segment of fly campaniform receptors. (A) Localization of Patronin-RFP (patronin-rfp-KI) in haltere receptors (lateral view). White arrowhead, Patronin-RFP in the TB. Inset (top view), distal (upper) and proximal (lower) ends of the MO (patronin-rfp-KI; dcx-emap-gal4/uas-cd4-tdgfp). Yellow arrowhead, two dot-shaped Patronin-RFP signals outside the neuron (Fig. S1). Scale bar, 1 µm. (B) Localization of Patronin-RFP in leg receptors (lateral view). White arrowhead, the distal signal of Patronin-RFP (inset, 600 nm from the distal tip). Blue arrowhead, the proximal signal of Patronin-RFP. Yellow arrowheads, two dot-shaped Patronin-RFP signals outside the neuron (Fig. S1). Scale bar, 1 µm. (C) EB1 comets (EB1-tdgfp-KI) in leg receptors. Left panel: A snapshot of EB1 comets (Video 2). White arrow, MO. Yellow arrowheads, two dot signals associated with the microtubules outside the neuron. Right panel: The trajectories of EB1 comets in 3 min. White arrowhead: dendritic inner segment (d). Scale bars, 5 µm. (D) Intensity line profile of EB1 signal in the outer segment along the proximal–distal axis (inset). The signals at the distal end (Id) and at the proximal part (Ip) were quantified. Scale bar (inset), 3 µm. (E) Statistical quantification of EB1 signal at the distal end (Id) and the proximal end (Ip; n = 5 cells). The data were presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. ***, P < 0.001. (F) Three sets of representative confocal images (−1 s: right before bleaching; 0 s: just after bleaching; 3 h or 9 min: time after bleaching) showing the recovery of GFP-tubulin (dcx-emap-gal4/uas-gfp-αtub84B) or EB1-GFP (dcx-emap-gal4; uas-EB1-gfp). Scale bars, 5 µm. (G) Fluorescence recovery curves after photobleaching. Red (circle), GFP-tubulin recovery in the outer segment (n = 10 cells). Black (square), GFP-tubulin recovery in the inner segment (n = 9 cells). Blue (triangle), EB1-GFP recovery in the outer segment (n = 6 cells). Data were presented as mean ± SEM. (H) Cartoon schematic of four groups of microtubules. 1, microtubules in the MO. 2, microtubules in the TB. 3, microtubules in the inner segment. 4, microtubules in the thecogen cell. +, plus end; −, minus end; BB, basal body; EDM, electron-dense materials; IS, inner segment; OS, outer segment; mem, membrane.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal