Figure 4.
Kat-60L1 contributes to mechanosensation. (A) Cartoon schematics of kat-60L1 and its two isoforms. The promoter regions: (1) long and short isoforms (P): kat-60L1-gal4; (2) long isoform (PL): kat-60L1-long-gal4; (3) short isoform (PS): kat-60L1-short-gal4. (B) Kat-60L1 expressed in fly external sensory organs and larval da neurons (kat-60L1-gal4; uas-cd4-tdgfp). c, Capitellum; p, pedicel; s, scabellum; a, antenna segment. Arrowheads, larval da and bipolar dendrites neurons (middle bottom) and larval ES organs (right bottom). Scale bars, 50 µm. Note the representative images for different tissues were juxtaposed together for display. (C) A representative gel image (RT-PCR) showing that the long isoform, but not the short isoform, can be detected in haltere RNA sample, while both isoforms can be detected in brain RNA sample. M, marker; S, short isoform; L, long isoform; hal, haltere. (D) Localization of RFP-kat-60L1-long in leg receptors. Upper: dcx-emap-gal4/uas-rfp-kat-60L1-long; +/uas-cd4-tdgfp. Lower: elav-gal4/uas-rfp-kat-60L1-long; +/uas-cd4-tdgfp. Scale bar, 5 µm. (E) Flight tests on wild type (n = 150), c01236 (n = 134), c01236/BE6 (n = 160), BE6 (n = 154), two rescue strains (1: elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6 [n = 132]; 2: dcx-emap-gal4/uas-RFP-kat-60L1-long; c01236/BE6 [n = 106]) and nompC3 (a negative control with abrogated mechanotransduction, n = 6). The number of flies is indicated. Two-sided unpaired Student’s t test with Bonferroni correction. ***, P < 0.001. (F) Representative extracellular recording traces of wild type, c01236/BE6, the rescue strain (elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6.), nompC3, and f02655. Note that f02655 is a dcx-emap mutant with compromised cytoskeleton in the outer segment (Bechstedt et al., 2010) and is used as a negative control here. (G) Statistical quantification of the response amplitude in extracellular recording assays. Wild type (n = 14), c01236/BE6 (n = 20), rescue: elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6 (n = 6), nompC3 (n = 4), and f02655 (n = 5). (H) Representative images of leg receptor in the functional assays. Upper: GCamp6s. Lower: Membrane. Yellow dashed circle, the region being compressed by the force probe. Arrowhead, soma. Scale bar, 10 µm. (I) Representative images of in vivo calcium recording in wild type (dcx-emap-gal4, uas-cd4-tdTom/uas-GCamp6s) and c01236/BE6 (dcx-emap-gal4, uas-cd4-tdTom/uas-GCamp6s; c01236/BE6). Scale bar, 10 µm. (J) Statistical quantification of the calcium response amplitude. Wild type (n = 18), c01236/BE6 (n = 17). In G and J, the number of flies (G) or cells (J) is indicated. Data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. ***, P < 0.001. BB, basal body; n.s., no significance.

Kat-60L1 contributes to mechanosensation. (A) Cartoon schematics of kat-60L1 and its two isoforms. The promoter regions: (1) long and short isoforms (P): kat-60L1-gal4; (2) long isoform (PL): kat-60L1-long-gal4; (3) short isoform (PS): kat-60L1-short-gal4. (B) Kat-60L1 expressed in fly external sensory organs and larval da neurons (kat-60L1-gal4; uas-cd4-tdgfp). c, Capitellum; p, pedicel; s, scabellum; a, antenna segment. Arrowheads, larval da and bipolar dendrites neurons (middle bottom) and larval ES organs (right bottom). Scale bars, 50 µm. Note the representative images for different tissues were juxtaposed together for display. (C) A representative gel image (RT-PCR) showing that the long isoform, but not the short isoform, can be detected in haltere RNA sample, while both isoforms can be detected in brain RNA sample. M, marker; S, short isoform; L, long isoform; hal, haltere. (D) Localization of RFP-kat-60L1-long in leg receptors. Upper: dcx-emap-gal4/uas-rfp-kat-60L1-long; +/uas-cd4-tdgfp. Lower: elav-gal4/uas-rfp-kat-60L1-long; +/uas-cd4-tdgfp. Scale bar, 5 µm. (E) Flight tests on wild type (n = 150), c01236 (n = 134), c01236/BE6 (n = 160), BE6 (n = 154), two rescue strains (1: elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6 [n = 132]; 2: dcx-emap-gal4/uas-RFP-kat-60L1-long; c01236/BE6 [n = 106]) and nompC3 (a negative control with abrogated mechanotransduction, n = 6). The number of flies is indicated. Two-sided unpaired Student’s t test with Bonferroni correction. ***, P < 0.001. (F) Representative extracellular recording traces of wild type, c01236/BE6, the rescue strain (elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6.), nompC3, and f02655. Note that f02655 is a dcx-emap mutant with compromised cytoskeleton in the outer segment (Bechstedt et al., 2010) and is used as a negative control here. (G) Statistical quantification of the response amplitude in extracellular recording assays. Wild type (n = 14), c01236/BE6 (n = 20), rescue: elav-gal4/uas-rfp-kat-60L1-long; c01236/BE6 (n = 6), nompC3 (n = 4), and f02655 (n = 5). (H) Representative images of leg receptor in the functional assays. Upper: GCamp6s. Lower: Membrane. Yellow dashed circle, the region being compressed by the force probe. Arrowhead, soma. Scale bar, 10 µm. (I) Representative images of in vivo calcium recording in wild type (dcx-emap-gal4, uas-cd4-tdTom/uas-GCamp6s) and c01236/BE6 (dcx-emap-gal4, uas-cd4-tdTom/uas-GCamp6s; c01236/BE6). Scale bar, 10 µm. (J) Statistical quantification of the calcium response amplitude. Wild type (n = 18), c01236/BE6 (n = 17). In G and J, the number of flies (G) or cells (J) is indicated. Data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. ***, P < 0.001. BB, basal body; n.s., no significance.

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