Figure 4

Larval sensory neurons contain centrioles but have compromised centriole function. yw (control) and DSas-4 embryos were fixed and stained with antibodies against either 22C10 to label stable microtubules or fluorescein isothiocyanate (FITC)-conjugated HRP to label neurons, β-galactosidase (to label heterozygote animals with a lacZ-marked balancer), and centriole-associated proteins. Stage 16 embryos were examined except where noted. (A) yw flies have punctate DSas-4 staining in ciliated neurons of the dorsal da cluster (arrows) and chordotonal neurons (box). DSas-4 embryos do not have any punctate DSas-4 staining. (B) Embryos were stained with FITC-conjugated HRP and γ-tubulin antibody. The γ-tubulin antibody localized at the base of each cilium in chordotonal neurons (boxes) of DSas-4 and yw animals. (C) Embryos were stained with antibodies against futsch (22C10) and the centriole-associated Drosophila pericentrin-like protein (D-PLP). D-PLP puncta were present in the chordotonal neurons of both control and DSas-4 mutants (boxes). Other centriolar proteins examined are seen in Additional file 6. (D) Cilia of chordotonal neurons were analyzed in DSas-4 and control animals. Top: embryos were stained with FITC-conjugated HRP. Middle: larvae expressing EB1-GFP in all neurons were examined by live imaging. Bottom: quantification of ciliary defects was performed. Neurons with abnormal cilia were classified as such by displaying either no cilium (arrow in (B), and arrow in the middle image of (D)) or a truncated cilium (asterisks, top image). DSas-4 mutants had a greater percentage of neurons with abnormal cilia compared to control samples. (E) Embryos were stained with antibodies against futsch and NompC. In control neurons, NompC localized to the distal tip of of mechanosensory cilia (arrow), while in DSas-4 mutants, NompC localization was seen at the base of the cilium (arrowhead).