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synj1 is mutated in comet mutants. A) Splice diagram of the zebrafish synj1 gene indicating alternate splicing of exons 13, 14 and 15, which can lead to the inclusion or exclusion of other indicated sequences (brackets). The diagram was assembled from collective synj1 sequencing data. Three new mutations of synj1 include deletion of the translation start site (donor splice site inactivation), and nonsense mutations, resulting in truncations at amino acid 296, or amino acid 943. A color-coded asterisk above the splice diagram indicates the position of the respective mutation in the transcript, and the expected protein product is depicted after the allele identifier, followed by the mutated nucleotide sequence. B,C) Expression of synj1 in inner ear and neuromast hair cells. B) synj1 expression pattern in the developing larva as shown by in situ hybridization. At 120 hpf, synj1 is expressed at high levels in the brain, eye and inner ear (arrow; upper panel). Hair-cell specific expression (asterisk) was observed in transverse thick sections of the ear at 100x magnification; both anti-sense synj1 and sense control label are shown (left and right lower panels, respectively). C) RT-PCR for synj1 from isolated neuromast hair-cells amplified the short (synj1-145) isoform. The panel depicts, from left to right, four control reactions (no template) and four hair-cell mRNA template reactions. gapdh mRNA was used as a positive control (band at 151 bp). Specific primer pairs were used to selectively amplify the long and short isoforms sj170 (101 bp expected band size) and sj145 (200 bp), respectively. The sj1 indicates total synj1 mRNA (140 bp).
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