Community Action Needed: Please respond to the NIH RFI
ZFIN ID: ZDB-FIG-190718-49
Marra et al., 2019 - Prostaglandin signaling regulates renal multiciliated cell specification and maturation. Proceedings of the National Academy of Sciences of the United States of America   116(17):8409-8418 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: 26+ somites
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: 26+ somites

Fig. 1

Chemical screen reveals prostaglandin signaling mediates renal MCC development in the zebrafish embryonic kidney, the pronephros. (A, Top) Schematic of a WT zebrafish embryo and pronephros at the 28 ss (lateral view) that is composed of a series of distinct functional segments with transporter cells (shades of orange) and MCCs (purple dots). The pronephros is situated along the trunk, lateral with respect to the adjacent somitic mesoderm and dorsal to the yolk and yolk sac extension. (A, Bottom) Somite map (indicated by the numbered box sequence) demarcates the location of nephron segments with respect to somites numbered 3 to 18. N, neck; P, podocyte. The formation of MCCs (purple dots) occurs in the central region of the nephron tubule, mostly located in the PST, with a few cells in the PCT and DE segments as well. (B, Top) Whole-mount smiFISH on a representative WT 28 ss embryo showing labeling for the presence of odf3btranscripts (magenta) and cdh17 transcripts (green). Nuclei are labeled with DAPI (blue), and the nephron is outlined by orange lines. (B, Bottom) Pseudocolored nuclei indicate scoring as transporter cell with cdh17+ only (green) or MCC with coexpression of cdh17+/odf3b+ (purple). (Scale bar, 10 μm.) (C) MCCs of a representative 28 ss WT embryo visualized by FISH, to label odf3b transcripts (magenta), and IF to label cilia with antiacetylated α-tubulin (white) and basal bodies with γ-tubulin (cyan). MCCs (circled in orange) exhibit odf3b expression and are distinguished by multiple ciliated basal bodies. (Scale bar, 10 μm.) (DF) Embryos (lateral view) were labeled by WISH for the MCC marker odf3b (purple), and in D and F for the DL marker slc12a3 (orange). (DF, Insets) Magnified dorsal views of the areas demarcated by the black boxes. (Scale bar, 50 μm.) (G) Quantification of odf3b+ cells. Each dot on the graph represents the total odf3b+ MCC number in one embryo. Data are represented as mean ± SEM; significance was determined by ANOVA, where *P < 0.0001, **P < 0.05, n.s., not significant.

 
 

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cdh17 TU control 26+ somites pronephric duct ISH
odf3b TU control 26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU chemical treatment by environment: indometacin 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU chemical treatment by environment: NS-398 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU chemical treatment by environment: SC560 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs1 control 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs1 chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs1 + MO1-ptgs2a control 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs1 + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs2a control 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
TU + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct ISH
26+ somites pronephric duct multi-ciliated epithelial cell ISH
slc12a3 TU control 26+ somites pronephric distal late tubule ISH
TU chemical treatment by environment: indometacin 26+ somites pronephric distal late tubule ISH
TU chemical treatment by environment: NS-398 26+ somites pronephric distal late tubule ISH
TU chemical treatment by environment: prostaglandin E2 26+ somites pronephric distal late tubule ISH
TU chemical treatment by environment: SC560 26+ somites pronephric distal late tubule ISH
TU + MO1-ptgs1 chemical treatment by environment: prostaglandin E2 26+ somites pronephric distal late tubule ISH
TU + MO1-ptgs1 + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric distal late tubule ISH
TU + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric distal late tubule ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
TU chemical treatment by environment: indometacin 26+ somites pronephric distal late tubule slc12a3 expression decreased amount, abnormal
26+ somites pronephric distal late tubule slc12a3 expression decreased distribution, abnormal
26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU chemical treatment by environment: NS-398 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU chemical treatment by environment: prostaglandin E2 26+ somites pronephric distal late tubule slc12a3 expression decreased amount, abnormal
26+ somites pronephric distal late tubule slc12a3 expression decreased distribution, abnormal
TU chemical treatment by environment: SC560 26+ somites pronephric distal late tubule slc12a3 expression decreased amount, abnormal
26+ somites pronephric distal late tubule slc12a3 expression decreased distribution, abnormal
26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU + MO1-ptgs1 control 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU + MO1-ptgs1 chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, ameliorated
TU + MO1-ptgs1 + MO1-ptgs2a control 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU + MO1-ptgs1 + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, ameliorated
TU + MO1-ptgs2a control 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, abnormal
TU + MO1-ptgs2a chemical treatment by environment: prostaglandin E2 26+ somites pronephric duct odf3b expression decreased distribution, abnormal
26+ somites pronephric duct multi-ciliated epithelial cell decreased amount, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA