Figure 1:
ref-2(
ot762) affects neuron development. (A) Fluorescent micrographs of
unc-53::gfp (DA and AS) and
unc-47::mCherry (DD and VD) in L4 wild-type control and
ot762 mutant animals, with ventral nerve cord motor neurons indicated by asterisks (scale bar 50µm in all panels). (B) Quantification of the number of VNC neurons expressing
unc-53::gfp and
unc-47::mCherry in controls and
ot762 mutants at L1 and L4 stages (mean ± standard deviation). (C) Confocal micrographs of
rab-3::tagRFP (all VNC neurons) in L4 control and
ot762 mutant. The average number of
rab-3::tagRFP(+) cells in the central region of the VNC decreases from 42.3 to 21. (D) Amino acid sequence of C. elegans of the fourth and fifth Zn finger of REF-2 aligned with orthologs from other species and location of amino acid substitutions caused in
ot327 (Let = lethal phenotype) and
ot762 (Unc = uncoordinated phenotype) alleles indicated below sequence. Hs = Homo sapiens, Ci = Ciona intestinalis, Dm - Drosophila melanogaster, Hv = Hydra vulgaris (Hv) , Ce = C. elegans. Blue indicates zinc coordinating cysteine and histidine residues, grey indicates complete conservation in species shown. (E) Fluorescent micrographs of
ttx-3::gfp (AIY interneuron) in L1 control and
ot762 mutant. (F) Quantification of the percent of worms with
ttx-3::gfp expression in AIY in controls and
ot762 mutants at the L1 and L4 stages (mean ± standard deviation). In all images, anterior is to the left.