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ZFIN ID: ZDB-FIG-190715-3
Tomar et al., 2019 - Blockade of MCU-Mediated Ca2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation. Cell Reports   26:3709-3725.e7 Full text @ Cell Rep.
PHENOTYPE:
Fish:
Observed In:
Stage: Adult

Fig. 3

Loss of MCU Limits mCa2+ Uptake and Elevates Total Body Fat in Zebrafish

(A) Schematic of the generation of global MCU KO zebrafish using CRISPR/Cas9.

(B) Genotyping for MCU deletion.

(C) Bar graph showing MCU mRNA abundance in WT, MCU+/−, and MCU−/− zebrafish.

(D) Western blot for MCU expression. n = 3.

(E) Measurement of mCa2+ uptake. n = 5–10.

(F) Quantification of ionomycin-induced peak mCa2+ levels from (E). n = 5–10.

(G) Bar graph representing cellular ATP levels in cells isolated from MCU+/+, MCU+/−, and MCU−/− zebrafish. n = 4.

(H) Adult WT and MCU−/− zebrafish were homogenized and centrifuged. MCU−/− zebrafish samples show a clear yellow color lipid layer on top of the protein lysate. n = 6.

(I) Adult WT and MCU−/− zebrafish were stained with Lipid Green to monitor the distribution of lipids in the whole body. n = 3.

(J and K) Bar graphs represent the quantification of Lipid Green staining from the dorsal (J) or tail (K) fin. n = 3.

Statistical analysis: mean ± SEM. p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Gene Expression Details No data available
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
mcutpl124/+ standard conditions Adult calcium import into the mitochondrion disrupted, abnormal
Adult cell ATP decreased amount, abnormal
Adult regulation of mitochondrial membrane potential process quality, normal
Adult whole organism lipid increased amount, abnormal
mcutpl124/tpl124 standard conditions Adult calcium import into the mitochondrion disrupted, abnormal
Adult caudal fin fat increased amount, abnormal
Adult cell ATP decreased amount, abnormal
Adult dorsal fin subcutaneous fat increased amount, abnormal
Adult regulation of mitochondrial membrane potential process quality, normal
Adult whole organism lipid increased amount, 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.