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. Author manuscript; available in PMC: 2017 Apr 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2016 Jan 28;36(4):655–662. doi: 10.1161/ATVBAHA.115.307087

Fig 2. The y58 and y68 mutations are defective in threonyl tRNA synthetase (tarsy58) and isoleucyl tRNA synthetase (iarsy68), respectively.

Fig 2

(A,B) Whole mount in situ hybridization of 24 hpf embryos probed for threonyl tRNA synthetase (tars, panel A) and isoleucyl tRNA synthetase (iars, panel B), showing expression in the somites. (C–E) Confocal fluorescence micrographs of the mid-trunk vasculature of 2 dpf Tg(fli1a-EGFP)y1 transgenic tarsy58−/− mutant larvae that were either un-injected (C), injected with Tol2(beta-actin:mCherry) transgene DNA (D), or injected with Tol2(beta-actin:tarsWT-2A-mCherry) transgene DNA (E). (F–H) Confocal fluorescence micrographs of the mid-trunk vasculature of 3 dpf Tg(fli1a-EGFP)y1 transgenic iarsy68−/− mutant larvae that were either un-injected (F), injected with Tol2(beta-actin:mCherry) transgene DNA (G), or injected with Tol2(beta-actin:iarsWT-2A-mCherry) transgene DNA (H). Panels C–H show lateral views of the trunk, rostral to the left. (I) Quantitation of the number of ectopic trunk intersegmental vessel branch points observed in phenotypically wild type (WT) siblings (Column 1, N = 10), Tol2(beta-actin:mCherry)-injected tarsy58−/− mutants (Column 2, N = 7), Tol2(beta-actin:tarsWT-2A-mCherry)-injected tarsy58−/− mutants (Column 3, N = 4), Tol2(beta-actin:mCherry)-injected iarsy68−/− mutants (Column 4, N = 9), or Tol2(beta-actin:iarsWT-2A-mCherry)-injected iarsy68−/− mutants (Column 5, N = 10). (J–M) Confocal fluorescence micrographs of the dorsalmost cranial vasculature of 3 dpf Tg(kdrl:GFP)la116 uninjected (J,L), 2.5 ng of tars ATG morpholino-injected (K), or 2.5 ng of iars ATG morpholino-injected (M) animals. Panels J–M show dorsal views, rostral to the left. (N) Quantitation of the number of ectopic dorsal cranial vessel branch points observed in phenotypically wild type siblings (Column 1, N = 4), tarsy58−/− mutants (Column 2, N = 6), or wild type animals injected with 2.5 ng of tars ATG morpholino (Column 3, N = 6). (O) Quantitation of the number of ectopic dorsal cranial vessel branch points observed in phenotypically wild type siblings (Column 1, N = 3), iarsy68−/− mutants (Column 2, N = 5), or wild type animals injected with 2.5 pg of iars ATG morpholino (Column 3, N = 5). Ectopic cranial branch points are quantitated at 48 hpf in tars mutants and morphants, and at 72 hpf in iars mutants and morphants. Scale bars = 100 μm. Significance at P < 0.05 is noted with an asterisk in panels I, N, O (NS = not significant).