ZFIN ID: ZDB-FIG-210715-5
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 Reagents:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 4 ppargc1a and Prostaglandin Pathway Component ptgs1 Act Together to Properly Form Ciliated Cells

(A) Twenty-eight hours post-fertilization WT (top), ppargc1a suboptimal dose MO (SMO), ptgs1 SMO, ppargc1a + ptgs1 SMO, and ppargc1a + ptgs1 SMO + dmPGE2-treated 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, 7 μm.

(B) Cilia length in micrometers for the proximal pronephros.

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

(D) Twenty-eight hours post-fertilization WT (top), ppargc1a SMO, ptgs1 SMO, ppargc1a + ptgs1 SMO, and ppargc1a + ptgs1 SMO + dmPGE2-treated 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, 7 μm.

(E) Cilia length in micrometers for the distal pronephros.

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

(G) MCCs stained via WISH for odf3b at 24 hpf in WT, ppargc1a SMO, ptgs1 SMO, ppargc1a + ptgs1 SMO, and ppargc1a + ptgs1 SMO + dmPGE2-treated zebrafish. Scale bar, 50 μm.

(H) Graph depicting the number of MCCs present.

(I) Forty-eight hours post-fertilization zebrafish injected with ppargc1a SMO, ptgs1 SMO, ppargc1a + ptgs1 SMO, and ppargc1a + ptgs1 SMO + dmPGE2-treated displaying either a WT phenotype (left) or an affected phenotype (right). The respective penetrance is listed within each image area. Scale bar, 100 μm.

Data are represented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (one-way ANOVA); 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 + MO1-ptgs1 standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
TU + MO1-ppargc1a + MO1-ptgs1 chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros multi-ciliated epithelial cell ISH
TU + MO1-ptgs1 standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC TU + MO1-ppargc1a standard conditions Prim-5 pronephros cilium
IHC TU + MO1-ppargc1a + MO1-ptgs1 standard conditions Prim-5 pronephros cilium
IHC TU + MO1-ppargc1a + MO1-ptgs1 chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros cilium
IHC TU + MO1-ptgs1 standard conditions Prim-5 pronephros cilium
Ab2-tubg1 IHC TU + MO1-ppargc1a standard conditions Prim-5 pronephros ciliary basal body
IHC TU + MO1-ppargc1a + MO1-ptgs1 standard conditions Prim-5 pronephros ciliary basal body
IHC TU + MO1-ppargc1a + MO1-ptgs1 chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros ciliary basal body
IHC TU + MO1-ptgs1 standard conditions Prim-5 pronephros ciliary basal body
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO1-ppargc1a standard conditions Prim-5 pronephros has fewer parts of type pronephros multi-ciliated epithelial cell, abnormal
Long-pec trunk curved ventral, abnormal
TU + MO1-ppargc1a + MO1-ptgs1 standard conditions Prim-5 pronephros cilium assembly decreased process quality, exacerbated
Prim-5 pronephros cilium decreased length, exacerbated
Prim-5 pronephros has fewer parts of type ciliated epithelial cell ciliary basal body, exacerbated
Long-pec trunk curved ventral, exacerbated
TU + MO1-ppargc1a + MO1-ptgs1 chemical treatment by environment: 16,16-dimethylprostaglandin E2 Prim-5 pronephros cilium assembly process quality, ameliorated
Prim-5 pronephros cilium length, ameliorated
Prim-5 pronephros has number of pronephros multi-ciliated epithelial cell, ameliorated
Long-pec trunk curvature, ameliorated
TU + MO1-ptgs1 standard conditions Prim-5 pronephros has fewer parts of type pronephros multi-ciliated epithelial cell, abnormal
Long-pec trunk curved ventral, abnormal
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.