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Fig. 1
Validation of pth2sa23129 fish lacking a functional peptide
(A) The pth2 gene consists of three exons; the coding sequence (CDS) is contained in exon 2 and 3. In pth2sa23129 fish, a T/A point mutation induces a premature stop codon within the propeptide sequence, thus preventing the successful translation and cleavage of the Pth2 peptide.
(B) Homozygous mutants were validated by sequencing and the premature ochre stop codon was identified at base pair 165 out of 474 of the CDS.
(C) Pth2+ cells are only found at the lateral edge of the dorsal thalamus, imaged here in a dorsoventral orientation. Transcript levels of pth2 were strongly decreased in pth2−/− animals, as demonstrated by fluorescent in-situ hybridization, although some remaining signal was detected (arrowheads). Abolished Pth2 translation was validated using a custom antibody recognizing a 27-mer epitope immediately preceding the peptide sequence (Anneser et al., 2020). For both wildtype and pth2−/−, 6 larvae at 6 dpf were imaged. Scale bar: 10 μm. R: rostral, (C) caudal. See also Figure S1