header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-150318-8
Gerlach et al., 2014 - Zebrafish pronephros tubulogenesis and epithelial identity maintenance are reliant on the polarity proteins Prkc iota and zeta. Developmental Biology   396(2):183-200 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-25 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-25 to Protruding-mouth

Fig. 3 prkCI and prkcζ are partially redundant during tubule polarity establishment, while prkCI is required for normal glomerulus development. (A) MO injections were performed with wild-type embryos to generate single prkCI, prkcζ, and double prkCI knockdowns, and morphants were scored at 72 hpf for gross morphology compared to embryos injected with control double prkCI 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, prkCI, prkcζ and prkCI morphant nephrons at the 36 hpf stage. Single morphants showed expression and proper localization of prkCI and p-ERM to the apical surface, whereas prkCI and p-ERM were absent in the double prkCI morphants. Na+/K+ ATPase was basolateral in wild-type embryos and single prkc morphants, but diffuse in the double prkCI/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 prkCI 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 prkCI morphants had disrupted PCT coiling morphogenesis. (Left columns, lateral view; right columns, dorsal view.)

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
slc20a1a TU control Protruding-mouth pronephric proximal convoluted tubule ISH
TU + MO1-prkci standard conditions Protruding-mouth pronephric proximal convoluted tubule ISH
TU + MO1-prkcz standard conditions Protruding-mouth pronephric proximal convoluted tubule ISH
wt1a TU control Protruding-mouth pronephric podocyte ISH
TU + MO1-prkci standard conditions Protruding-mouth pronephric podocyte ISH
TU + MO1-prkcz standard conditions Protruding-mouth pronephric podocyte ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab1-atp1a1 IHC TU control Prim-25 pronephros basolateral plasma membrane
IHC TU + MO1-prkci standard conditions Prim-25 pronephros basolateral plasma membrane
IHC TU + MO1-prkci + MO1-prkcz standard conditions Prim-25 pronephros apical plasma membrane
IHC Prim-25 pronephros basolateral plasma membrane
IHC TU + MO1-prkcz standard conditions Prim-25 pronephros basolateral plasma membrane
Ab1-ezr IHC TU control Prim-25 pronephros apical plasma membrane
IHC TU + MO1-prkci standard conditions Prim-25 pronephros apical plasma membrane
IHC TU + MO1-prkci + MO1-prkcz standard conditions Prim-25 Not Detected pronephros apical plasma membrane
IHC Prim-25 Not Detected pronephros basolateral plasma membrane
IHC TU + MO1-prkcz standard conditions Prim-25 pronephros apical plasma membrane
Ab1-prkcz IHC TU control Prim-25 pronephros apical plasma membrane
IHC TU + MO1-prkci standard conditions Prim-25 pronephros apical plasma membrane
IHC TU + MO1-prkci + MO1-prkcz standard conditions Prim-25 Not Detected pronephros apical plasma membrane
IHC Prim-25 Not Detected pronephros basolateral plasma membrane
IHC TU + MO1-prkcz standard conditions Prim-25 pronephros apical plasma membrane
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO1-prkci standard conditions Protruding-mouth glomerular visceral epithelial cell migration decreased process quality, abnormal
Protruding-mouth pronephric podocyte mislocalised laterally, abnormal
Protruding-mouth pronephric proximal convoluted tubule malformed, abnormal
Protruding-mouth pronephric proximal tubule morphogenesis decreased process quality, abnormal
TU + MO1-prkci + MO1-prkcz standard conditions Prim-25 pronephric tubule establishment of epithelial cell apical/basal polarity decreased occurrence, abnormal
TU + MO1-prkcz standard conditions Protruding-mouth glomerular visceral epithelial cell migration process quality, normal
Protruding-mouth pronephric podocyte position, normal
Protruding-mouth pronephric proximal convoluted tubule morphology, normal
Protruding-mouth pronephric proximal tubule morphogenesis process quality, normal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 396(2), Gerlach, G.F., Wingert, R.A., Zebrafish pronephros tubulogenesis and epithelial identity maintenance are reliant on the polarity proteins Prkc iota and zeta, 183-200, Copyright (2014) with permission from Elsevier. Full text @ Dev. Biol.