Laterality defects of the brain and heart in zebrafish embryos. (A) Ventra lviews of the parapineal (arrow) and the myocardium in 48 hpf Tg(foxd3:EGFP/myl7:EGFP) embryos (a–e) visualized using 3D SPIM, which revealed their laterality defect at the cellular level. 10× lens; 1.5× zoom; scale bar, 50 µm. (B) Dorsal views of left-right asymmetry of the pineal complex (a–d) and ventral views of the heart (a’–d’) were imaged at 4 days post-fertilization using a fluorescent stereomicroscope. Scale bar, 50 µm. (C) Embryos treated with and without cold shock at 22.5°C (n = 226) and 28.5°C (n = 795), respectively, were examined for their laterality defects of the parapineal (arrow) and cardiac looping at 72hpf. LD, left parapineal(L)and dextral heart looping (D); LL, left parapineal(L)and leftward heart looping (L); RD, right parapineal (R) and dextral heart looping (D); RL, right parapineal (R) and leftward heart looping (L); NU, no parapineal (N) and unloop heart (U); pp, parapineal; R-L, right-left sides; L-R, left-right sides; A, atrium; V, ventricle. *P< .05, **P< .01, and ***P< .001 by Student’s t-test.
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