Human AL-LC induced functional, biochemical, and ultrastructural alterations in the hearts of the transgenic fish. Representative images of ventricular area (VA) in diastole (VAd) and systole (VAs) obtained from color Doppler at 4 mo of age in control and transgenic fish (left). Quantification of fractional area change (FAC; top right). Heart rate of control and transgenic fish during acquisition (bottom right) (A). Peak aortic blood flow velocity at 4 mo in control and transgenic fish, n = 6 fish per experimental group (*P = 0.02; **P = 0.008) (B). Increased apoptosis was observed in the transgenic zebrafish at 3 mo old compared with control fish, as evidenced by the presence of TUNEL-positive nuclei in representative myocardial sections and an increase in cleaved caspase-3 detected by immunoblot of isolated heart protein lysates (P = 0.001) (C and D). Representative immunoblot showing increased LC3-II (P = 0.02) expression in transgenic myocardium compared with controls at 1 yr of age is indicative of abnormal authophagic flux (E). Representative images of VA in VAd and VAs obtained from color Doppler at 20 mo of age in control and transgenic fish (left). Quantification of FAC (top right). Heart rate of control and transgenic fish during acquisition (bottom right) (F). Peak aortic blood flow velocity at 20 mo in control and transgenic fish, n = 9 (control), n = 13 (transgenic) (*P = 0.02; **P = 0.009) (G). Scatter plot showing negative correlation between heart rate and aortic flow rate in transgenic fish (P = 0.06) (H). bpm, beats/min; Con, control; TG, transgenic.
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