Fig. 2

Effects of zlrrc10-MO1 on the zebrafish heart morphology and blood flow. (A) The effect of zlrrc10-MOs on blocking translation of zlrrc10. zlrrc10 cDNA yielded a [³⁵S]-methionine labeled 31 kDa protein by in vitro transcription and translation reaction (+). Negative control (-) was incubated without template plasmid. To determine the effect of zlrrc10-MOs, zlrrc10 cDNA was in vitro translated in the presence of MO1 or MO2. To confirm the specificity of zlrrc10 MOs, two control morpholinos, CMO or CMO-5m were used. The final concentrations of morpholinos in the reaction are indicated. (B) Knockdown of Lrrc10 caused heart defects. CMO (a and d) and MO1 (b, c, e, and f) injected zebrafish were imaged at 3 dpf. d–f are high-power images of a–c, respectively, which show heart looping defects of the morphants (e and f). The arrowhead indicates a large pericardial edema. Blue and red indicate the ventricle and atrium, respectively. A, atrium; V, ventricle. (C) Reduced blood flow in the morphants. Zebrafish embryos were injected with zlrrc10-MO1 (hatched bar) or CMO (open bar). RBC perfusion rates in an intersegmental vessel were determined between 2 and 5 dpf as described in Materials and methods section. An asterisk indicates a significant difference in values between the test CMO and the MO1 at p < 0.05.