Quantification of urinary excretion in zebrafish. (A) After pericardial Alexa 488 injections, serial images were taken of zebrafish urinary structures. Scale bar: 0.4 mm. Regions of interest were identified as follows: i, proximal bladder; ii, middle bladder; iii, distal bladder; iv, total bladder; v, urinary stream. vi and vii were selected for normalization of body and urinary streams, respectively. (B) Plots of total bladder (iv) fluorescence intensity were plotted alongside urinary stream (v) intensity over time. (C) The distribution of bladder emptying intervals (E.I.) in seconds was plotted. (D) Bladder subregion fluorescent intensity plotted against time reveals that simultaneous contraction, and not sequential peristalsis, empties the full urinary bladder.
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