Figure 2

Cleared mA?42 accumulates in the pronephros via blood flow. (A,B) mA?42-injected larvae after propranolol (100 ?M) treatment. The mA?42 intensity in the pronephros of propranolol-treated larvae (white dotted lines) decreased (B) compared to control (A). (C,D) mA?42-injected larvae after control morphants (C) and 200 ?M tnnt2a morphants (D). (E) Quantification of heartbeats upon propranolol treatment. n = 9 for control group, n = 10 for propranolol. (F) Quantification of the mA?42 intensity in the pronephros after propranolol treatment. n = 8 per group. (G) Quantification of the mA?42 intensity in the pronephros of control and tnnt2a morphants. Statistical significance was determined by two tailed unpaired t-test. Data are presented as mean ± SEM. n = 9 per group. Data are representative of at least three independent experiments. (H) A schematic diagram of experimental setting of A? and tracer injection. mA?42 or oA?42 (HiLyte) was injected into ventricle and 10 kDa Dextran was injected into caudal vein. (I) Quantification of the A?42 fluorescence intensity in the pronephros normalized by background fluorescence. n = 9 for mA?42, n = 8 for oA?42. Data are representative of at least three independent experiments. (J?K?) Confocal fluorescence images showing the brain and pronephros of zebrafish with lateral view after mA?42 (J) or oA?42 (K) injection at 3 dpf. Red fluorescence indicates A?42-HiLyte Fluor and green fluorescence show 10 kDa dextran tracer injected into caudal vein. (J?) Ventricle-injected mA?42 was seen in the pronephros region (white dotted lines and depicted as p). (K?) Ventricle-injected oA?42 was seen only in the brain region (b), but not detected in the pronephros. Caudal vein-injected dextran accumulated in the pronephros of both mA?42 and oA?42-injected larvae (J?,K?). (J?,K?) show merged images. b, brain; MO, morpholino; p, pronephros; Scale bars = 100 ?m. ** p < 0.001; *** p < 0.0005; **** p < 0.0001.