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. 2011 May 12;6(5):e19632. doi: 10.1371/journal.pone.0019632

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Mochizuki et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A, B) MBs in the wild-type and unc-51 ^25/25 mutant larvae at the late third instar stage. MBs were visualized with mCD8::GFP driven by 201Y. (C) Quantification of the dendrite overextension phenotype in wild-type and unc-51 mutant larvae. While 14% of the wild-type MBs (n = 21) exhibited short overextensions, 83% of the unc-51 ^−/− MBs (n = 24) exhibited short or long overextensions (shorter or longer than the diameter of the calyx, respectively). (D, E) Analyses of the dendritic phenotype using MARCM clones. Clones were induced by a heat shock at the early first instar stage. MB neurons were labeled with UAS-mCD8::GFP driven by elav ^c155. (D) Wild-type neuroblast clone. (E) unc-51 ^3/3 mutant neuroblast clone. (F) Quantification of the dendrite overextension phenotype in MARCM clones. While only 6% of the wild-type clones (n = 17) showed overextensions, all of the unc-51 ^3/3 clones (n = 23) exhibited short or long overextensions (shorter or longer than the diameter of the calyx, respectively). (G–J) Dendrite overextension phenotype in unc-51 mutant MBs was rescued by pan-neuronal expression of unc-51 ^WT but not by the kinase-deficient (K38A) unc-51 ^K38A transgene. (G) Wild-type. (H) unc-51 ^25/25 mutant. Rescue genotypes: (I) w; elav-GAL4, UAS-mCD8::GFP, UAS-unc-51 ^WT; unc-51 ^25/25 and (J) w; elav-GAL4, UAS-mCD8::GFP, UAS-unc-51 ^K38A; unc-51 ^25/25. (K) Quantification of the dendrite overextension phenotype. While none of the wild-type MBs (elav-GAL4, UAS-mCD8::GFP) (n = 6) exhibited calyx overextension, 83% of the unc-51 ^25/25 mutant MBs (elav-GAL4, UAS-mCD8::GFP; unc-51 ^25/25) exhibited short or long overextensions (n = 6). The dendritic overextension phenotype was significantly suppressed by the unc-51 ^WT transgene (w; elav-GAL4, UAS-mCD8::GFP, UAS-unc-51 ^WT; unc-51 ^25/25), with 31% of the MBs exhibiting short extensions (n = 16). Expression of unc-51 ^K38A failed to rescue the phenotype (w; elav-GAL4, UAS-mCD8::GFP, UAS-unc-51 ^K38A; unc-51 ^25/25), with 73% of the MBs exhibiting short or long overextensions (n = 22). Note different genetic background from that of the experiments in (A–C). Arrows in (B, E, H and J) indicate overextensions from the calyx. Scale bar, 10 µm.