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Fig. 5 | Journal of Biomedical Science

Fig. 5

From: A thrombomodulin-like gene is crucial to the collective migration of epibolic blastomeres during germ layer formation and organogenesis in zebrafish

Fig. 5

The impact of zTM-b knockdown in heart and blood cell formation. Wild-type and Tg (mpx:eGFP) embryos were injected with TM-b MO with/without co-injecting plasmids encoding human TM and examined for the development of heart and blood cells. a-d The cardiac ejection fraction and cardiac output of larvae at 3 dpf were calculated from the change in the dimension of cardiac chamber (the circled area in (a)) in the serial video still images (lateral view with anterior to the right) as described in Materials and Methods. e, f Embryos at 1 dpf were subjected to WISH for the distribution of cmlc2 transcripts, a cardiac primordium marker. Embryos displaying signal with decreased intensity or spatially aberrant distribution (heart tube with mid-line positioning or right-jogging) were categorized as abnormal and quantified. g, h The development of embryonic right-left patterning was evaluated at 12 hpf and categorized into normal, small, absent and multiple based on the number and morphology of Kupffer’s vesicle (arrows). i, j Embryos at 3 dpf were stained with o’-dianisidine for hemoglobin distribution. Larvae displaying weaker, dispersed or ectopic signals in common cardinal veins (arrowheads in left panel) and/or absence of posterior blood island (arrowheads in the right panel) were categorized as abnormal and quantified. k, l Tg (mpx:eGFP) embryos with/without TM-b knockdown were imaged at 2 dpf and scored from 1 to 5 points based on the intensity of green fluorescence, which represented the quantity of mpx-positive leukocytes (mainly neutrophils). Data were collected from at least 3 independent experiments with the total embryo numbers of 11–84 for each group. WT, un-injected wild-type embryos; MO, embryos injected with TM-b MO; hTM, human thrombomodulin; CTL, Tg (mpx:eGFP) embryos without MO injection; A, atrium; V, ventricle; EDA, end-diastolic area; ESA, end-systolic area

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