Fig. 3

prk_CI and prkcζ are partially redundant during tubule polarity establishment, while prk_CI is required for normal glomerulus development. (A) MO injections were performed with wild-type embryos to generate single prk_CI, prkcζ, and double prk_CI/ζ knockdowns, and morphants were scored at 72 hpf for gross morphology compared to embryos injected with control double prk_CI/ζ mismatch MOs. (Left) Percentage distributions of three classes of phenotypes, categorized as wild-type, mild, and severe. These categories were based on curled tail, edema, and patchy eye pigmentation (Right, lateral views of live representative embryos). (B) Confocal imaging of IF analysis during tubulogenesis in wild-type, prk_CI, prkcζ and prk_CI/ζ morphant nephrons at the 36 hpf stage. Single morphants showed expression and proper localization of prk_CI/ζ and p-ERM to the apical surface, whereas prk_CI/ζ and p-ERM were absent in the double prk_CI/ζ morphants. Na⁺/K⁺ ATPase was basolateral in wild-type embryos and single prkc morphants, but diffuse in the double prk_CI/ζ morphants. Scale bars, 5 µm. (C) WISH analysis of podocyte and PCT development in wild-type control embryos and single prkc morphants using wt1a and slc20a1a, respectively. prkcζ morphants had midline clusters of podocytes, similar to wild-type embryos, while prk_CI morphants had scattered clusters of podocytes that failed to migrate to the midline. PCT coiling morphogenesis was normal in prkcζ morphants and wild-type control embryos, while prk_CI morphants had disrupted PCT coiling morphogenesis. (Left columns, lateral view; right columns, dorsal view.)