ZFIN ID: ZDB-FIG-210715-4
Chambers et al., 2020 - Ppargc1a Controls Ciliated Cell Development by Regulating Prostaglandin Biosynthesis. Cell Reports   33:108370 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 3 ppargc1a Loss of Function Phenotypes Can Be Rescued by Supplementing with PGE2

(A) Relative concentration of PGE2 in WT and ppargc1a morphants.

(B) MCCs stained via WISH for odf3b at 24 hpf in WT (left), ppargc1a MO (middle), and ppargc1a morphants treated with dmPGE2 (right). Scale bar, 90 μm.

(C) Graph depicting the number of MCCs present in WT, ppargc1a MO, and ppargc1a MO treated with dmPGE2.

(D) Twenty-eight hours post-fertilization WT (top), ppargc1a MO-injected (middle), and ppargc1a MO-injected + dmPGE2 (bottom) zebrafish stained via whole-mount immunofluorescence for acetylated α-tubulin (cilia, green), γ-tubulin (basal bodies, red), and DAPI in the proximal pronephros. Scale bar, 10 μm.

(E) Cilia length in micrometers for the proximal pronephros of WT, ppargc1a morphants, and ppargc1a morphants treated with dmPGE2.

(F) Graph of the percentage of ciliated basal bodies/total basal bodies per 100 μm in the proximal pronephros.

(G) Twenty-eight hours post-fertilization WT (top), ppargc1a MO-injected (middle), and ppargc1a MO-injected + dmPGE2 (bottom) zebrafish stained via whole-mount immunofluorescence for acetylated α-tubulin (cilia, green), γ-tubulin (basal bodies, red), and DAPI in the distal pronephros. Scale bar, 10 μm.

(H) Cilia length in micrometers for the distal pronephros of WT, ppargc1a morphants, and ppargc1a morphants treated with dmPGE2.

(I) Graph of the percentage of ciliated basal bodies/total basal bodies per 100 μm in the distal pronephros.

(J and K) Scatterplots illustrating the difference in fluorescent intensity signals of WT, ppargc1a MO, and ppargc1a MO + dmPGE2 in proximal (J) and distal (K) cilia.

(L) Representative images of ppargc1a+/sa13186 in crosses shown at 48 hpf WT zebrafish treated with DMSO (top), curly ppargc1asa13186/sa13186 treated with DMSO (middle), and a WT-appearing, genotype-confirmed ppargc1asa13186/sa13186 rescued with dmPGE2 (bottom). Scale bar, 90 μm.

(M) The percentage of zebrafish with WT (black) or curly (white) phenotypes at 48 hpf after treatment with DMSO or dmPGE2. Chi-square test indicates significance.

Data are represented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (one-way ANOVA or t test); n.s., not significant.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
odf3b TU standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
TU + MO1-ppargc1a standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
TU + MO1-ppargc1a chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros multi-ciliated epithelial cell ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC TU standard conditions Prim-5 pronephros cilium
IHC TU + MO1-ppargc1a standard conditions Prim-5 pronephros cilium
IHC TU + MO1-ppargc1a chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros cilium
Ab2-tubg1 IHC TU standard conditions Prim-5 pronephric tubule ciliary basal body
IHC TU + MO1-ppargc1a standard conditions Prim-5 pronephric tubule ciliary basal body
IHC TU + MO1-ppargc1a chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephric tubule ciliary basal body
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO1-ppargc1a standard conditions Prim-5 pronephros cilium ab1-tuba labeling decreased amount, abnormal
Prim-5 pronephros cilium assembly decreased process quality, abnormal
Prim-5 pronephros cilium decreased length, abnormal
Prim-5 pronephros distal region has fewer parts of type ciliated epithelial cell ciliary basal body, abnormal
Prim-5 pronephros has fewer parts of type pronephros multi-ciliated epithelial cell, abnormal
Prim-5 pronephros proximal region has fewer parts of type ciliated epithelial cell ciliary basal body, abnormal
TU + MO1-ppargc1a chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros cilium ab1-tuba labeling amount, ameliorated
Prim-5 pronephros cilium assembly process quality, ameliorated
Prim-5 pronephros cilium length, ameliorated
Prim-5 pronephros distal region has number of ciliated epithelial cell ciliary basal body, ameliorated
Prim-5 pronephros has number of pronephros multi-ciliated epithelial cell, ameliorated
Prim-5 pronephros proximal region has number of ciliated epithelial cell ciliary basal body, ameliorated
ppargc1asa13186/sa13186(TU) control Long-pec post-vent region curved ventral, abnormal
Long-pec trunk curved ventral, abnormal
ppargc1asa13186/sa13186(TU) chemical treatment by environment: 16,16-dimethylprostaglandin E2 Long-pec post-vent region curvature, ameliorated
Long-pec trunk curvature, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.