Fig. 3. Lowered heart rate, ventricular dimensions, stroke volume and cardiac output in zebrafish embryos treated with cigarette smoke or snuff extracts.

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Fig. 3. Lowered heart rate, ventricular dimensions, stroke volume and cardiac output in zebrafish embryos treated with cigarette smoke or snuff extracts. Lowered heart rate, ventricular dimensions, stroke volume and cardiac output in zebrafish embryos treated with cigarette smoke or snuff extracts. (A) Still images from a microscope video of a zebrafish larvae at 3 days post-fertilization (dpf). The area indicated by a red box in the image to the left is enlarged and shown at end systole or end diastole in the middle or right images, respectively. A, Atrium; rl, long-axis radius; rs, short-axis radius; V, volume. (B,C) Quantification of the average volume of the ventricle at systole (B) or diastole (C) in 3 dpf embryos treated with nothing, cigarette smoke extract at 1:30 dilution (CS), snuff extract at 1:50 dilution (Sn) or the anesthetic tricaine (Tri) at 0.2 mg/ml. n=10, 3, 9 and 5 embryos (three measurements per embryo) in the control, CS, SN and Tri groups respectively. *P<0.05, ***P<0.001. (D) Quantification of the stroke volume in picoliters (pl) of the same 3 dpf embryos as in B and C. **P<0.01, ***P<0.001. (E) Quantification of the average heart rate in beats per minute (beats/min) of the same 3 dpf embryos as in B and C. **P<0.01, ***P<0.001. (F) Quantification of the cardiac output in nanoliters per minute (nl/min) of the same 3 dpf embryos as in B and C. **P<0.01, ***P<0.001. The experiment was repeated with similar results. Results are shown as means±s.e.m. and statistical evaluation was done using Student’s two-tailed t-test assuming equal variance between the groups. Maggie Folkesson et al. Biology Open 2016;5:970-978 © 2016. Published by The Company of Biologists Ltd