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Morphology of pronephros and interrenal tissue in the absence of blood flow. (A) The interrenal steroidogenic tissue positive for 3β-Hsd activity forms an extension that protrudes toward the midline by 54 hpf (cell protrusions marked by red arrows), while kidney glomeruli delineated by wt1b: GFP expression (G, yellow arrowheads) assemble at the midline. Morphogenetic movements of kidney glomeruli and steroidogenic tissues are defective in the tnnt2a morphant. All panels show dorsal views of representative embryos. (B) Quantification of effects of tnnt2aMO injection on interrenal migration. The distance between the midline and the migrating tip of steroidogenic tissue was designated as positive if the migrating tip had not reached the midline, and negative if the tip had migrated across the midline. (C) Relative density of steroidogenic tissue, as assessed by 3β-Hsd activity staining in the ventral surface, in tnnt2a morphants compared to wild-type controls. The number of embryos in each group is indicated in parentheses in (B) and (C). Histograms with different letters above them are significantly different (ANOVA and Duncan′s multiple test, P<0.05). (D) Effect of camptothecin treatment from 48 to 57 hpf on 3β-Hsd activity in steroidogenic cells. A, anterior; P, posterior; L, left; R, right. Broken yellow lines indicate position of the midline. Abbreviations: glomerulus (G). Scale bar, 50 µm.
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