ZFIN ID: ZDB-FIG-210715-3
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:
Genes:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-5

Fig. 2 Loss of ppargc1a Results in Ciliogenesis Defects

(A) Schematic indicating location of MCCs in the proximal portion and monociliated cells in the distal portion of the zebrafish pronephros at 24 hpf.

(B) WT (top) and ppargc1asa13186/sa13186 mutant (bottom) zebrafish at 24 hpf stained via WISH for odf3b to mark MCCs. Scale bar, 90 μm.

(C) Graph representing the number of MCCs present in WT and ppargc1asa13186/sa13186 zebrafish at 24 hpf.

(D) Fluorescent in situ hybridization for MCC marker (odf3b, green) and transporter cell marker (trpm7, red) in WT and ppargc1a morphant zebrafish at 24 hpf. Scale bar, 10 μm.

(E and F) Graphs representing the percentage of odf3b+ (E) or trpm7+ (F) cells in WT (black) and ppargc1a morphants (white) pronephros between somites 9 and 11.

(G) Twenty-eight hours post-fertilization WT (top), ppargc1a MO-injected (middle), and ppargc1asa13186/sa13186 (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.

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

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

(J) Twenty-eight hours post-fertilization WT (top) and ppargc1a MO-injected (middle) and ppargc1asa13186/sa13186 (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.

(K) Cilia length in micrometers for the distal (right) pronephros.

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

(M and N) Fluorescent intensity plots of WT (black) and ppargc1a MO-injected (red) zebrafish in proximal (M) and distal (N) pronephros. Data are represented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 (one-way ANOVA or t test).

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
ppargc1asa13186/sa13186(TU) standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
trpm7 TU standard conditions Prim-5 pronephros single ciliated epithelial cell ISH
TU + MO1-ppargc1a standard conditions Prim-5 pronephros single 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 ppargc1asa13186/sa13186(TU) standard conditions Prim-5 pronephros cilium
Ab2-tubg1 IHC TU standard conditions Prim-5 pronephros ciliary basal body
IHC TU + MO1-ppargc1a standard conditions Prim-5 pronephros ciliary basal body
IHC ppargc1asa13186/sa13186(TU) standard conditions Prim-5 pronephros 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 extra parts of type pronephros single ciliated epithelial cell, 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
ppargc1asa13186/sa13186(TU) standard conditions 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
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.