Fat body-specific reduction of CTPS alleviates HFD-induced obesity
- School of Life Science and Technology, ShanghaiTech University, China
- College of Life Sciences, Shanghai Normal University, China
- Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China
- University of Chinese Academy of Sciences, China
- Department of Physiology, Anatomy and Genetics, University of Oxford, United Kingdom
Figures
Figure 1 with 3 supplements
CTPS knockdown in the Drosophila fat body leads to body weight loss.
(A–C) Body weights of 5-day-old adult flies from the indicated genotypes (30 flies/group, 5–6 groups/genotype, 2–3 biological replicates). TubG4ts>CTPS-Ri versus TubG4ts>+, CTPS-Ri/+ or TubG4ts>Con-R…
Figure 1—figure supplement 1
Quantitative RT-PCR analysis.
(A–C) Quantitative RT-PCR analysis of the mRNA abundance of CTPS from whole-body (A, C) or head (B) lysates of adult flies in indicative lines (10 flies/group, 3 groups/genotype, 3 biological …
Figure 1—figure supplement 2
Inhibition of CTPS in adipocytes reduced body weight and resistance to starvation.
(A) Body weight of 5-day-old male adults of the indicated genotypes (30 flies/group, 3 groups/genotype, 2 biological replicates). pplG4>CTPS-Ri versus pplG4>+ or CTPS-Ri/+. All values are means ± …
Figure 1—figure supplement 3
TAG level of male adults upon food deprivation.
(A) The TAG levels of male adults from indicative lines. CgG4>CTPS-Ri versus CgG4>+, CTPS-Ri/+, and CgG4>Con-Ri line. TAG levels are normalized to protein level (10 flies/group; 5–6 groups/genotype, …
Figure 2 with 2 supplements
Adipocyte-specific knockdown of CTPS decreases larval body weight.
(A) The 3rd instar wandering larval body weight of the indicated lines (10 larvae/group, 5–6 groups/genotypes, 3 biological replicates). CgG4>CTPS-Ri is compared with CgG4>+, CTPS-Ri/+ or CgG4>Con-Ri…
Figure 2—figure supplement 1
Larval size comparison.
(A) 3rd instar wandering larvae from the indicated lines. (B) Quantification of larval size from (A) (10 larvae/group; 3 groups/genotype, 3 biological replicates). The value is normalized to the CgG4…
Figure 2—figure supplement 2
Quantitative RT-PCR analysis.
(A) Quantitative RT-PCR analysis of CTPS mRNA abundance in the fat body lysate of 3rd instar wandering larvae from the indicated lines (30 larvae/group; 4 groups/genotype; 2 biological replicates). …
Figure 3
HFD promotes CTPS expression in the fat body.
(A) Quantitative RT-PCR analysis of the abundance of CTPS mRNA in fat body lysates of 76~80 hour after egg laying (AEL) larvae under RD and HFD conditions. The relative value is normalized with …
Figure 4 with 3 supplements
Knockdown of CTPS in adipocytes alleviates HFD-induced obesity.
(A) 76~80 hour AEL larvae expressing eGFP (green) with CgG4 driving CTPS knockdown in the fat body and the wild-type control were fed with RD or HFD (eGFP fluorescent image top, bright-field image …
Figure 4—figure supplement 1
Quantitative RT-PCR analysis.
(A) Quantitative RT-PCR analysis of eGFP mRNA abundance in the fat body or whole body of 3rd instar wandering larvae fed HFD (30 larvae/group; 3 groups/genotype; 2 biological replicates). All data …
Figure 4—figure supplement 2
CTPS is required for adipocyte growth.
(A) Representative confocal images of fat bodies showing clones of adipocytes with CTPS knockdown (green) from a 3rd instar larva (2 biological replicates). The areas in white dotted boxes are …
Figure 4—figure supplement 3
The effects of CTPS on phospholipid composition and the expression of nucleotide diphosphate kinases.
(A) Relative level of phospholipid in the fat bodies of 76~80 hour AEL larvae. LPE, lysophophatidylethanolamine; LPA, lysophophatidic acid; LPC, lysophosphatidylcholine; LPS, lysophosphatidylserine; …
Figure 5 with 3 supplements
Fat-body-specific knockdown of CTPS reduces lipogenic gene expression.
(A) The fat bodies of 2nd instar larvae from CgG4>CTPS-Ri and CgG4>+ larvae were analyzed by RNA-seq analysis. A heat map of relative gene expression, obtained using RNA-seq data, is depicted for …
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Figure 5—source data 1
List of the differentially expressed genes in the heatmap.
Differentially expressed genes were identified on the basis of fold change values (gene expression level in the CgG4>CTPS-Ri relative to that in the CgG4>+ control line, ≥2.0-fold change, Student’s t-test, P<0.05).
- https://cdn.elifesciences.org/articles/85293/elife-85293-fig5-data1-v1.xls
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Figure 5—source data 2
Uncropped gel of phosphorylated Akt from fat body lysates.
Anti-mCh, anti-phosphorylated-Akt, and anti-total-Akt antibodies were used for the immunoblotting analysis. Alpha-tubulin was used as an internal control.
- https://cdn.elifesciences.org/articles/85293/elife-85293-fig5-data2-v1.zip
Figure 5—figure supplement 1
KEGG functional classification of the genes that are affected by CTPS knockdown.
(A) Scatter plot comparing transcript abundance between the CTPS knockdown larvae and the control. Differentially expressed genes were identified on the basis of the fold change values (gene …
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Figure 5—figure supplement 1—source data 1
List of baseMean values for the CgG4>CTPS-Ri line relative to the CgG4>+ control line.
- https://cdn.elifesciences.org/articles/85293/elife-85293-fig5-figsupp1-data1-v1.xls
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Figure 5—figure supplement 1—source data 2
Numbers of genes belonging to distinct functional groups that are up- or downregulated in the CgG4>CTPS-Ri line relative to the CgG4>+ control line.
Gene functions are predicted on the basis of the KEGG database.
- https://cdn.elifesciences.org/articles/85293/elife-85293-fig5-figsupp1-data2-v1.xls
Figure 5—figure supplement 2
Quantitative RT-PCR analysis.
(A–B) Quantitative RT-PCR analysis of acc and fasn1 mRNA abundance in the fat body lysates of 76~80 hour AEL larvae (30 larvae/group; 4 groups/genotype; 2 biological replicates). Actin5c (A) and rp32…
Figure 5—figure supplement 3
Quantitative RT-PCR analysis.
(A) Quantitative RT-PCR analysis of Pi3K mRNA abundance in the fat body lysates of 76~80 hour AEL larvae (30 larvae/group; 3 groups/genotype; 2 biological replicates). All data are shown as mean ± …
Figure 6 with 2 supplements
Disrupting the filament-forming property of CTPS alleviates HFD-induced obesity.
(A) Representative photograph of HFD-fed 76~80 hour AEL larvae showing larval morphology. The CgG4> CTPSMU OE line is compared with CgG4>+, CgG4>GFP, CgG4>CTPSWT-OE, CTPSWT-OE/+, and CTPSMU-OE/+ …
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Figure 6—source data 1
Uncropped gel of phosphorylated Akt from fat body lysates.
Anti-mCh, anti-phosphorylated-Akt, and anti-total-Akt antibodies were used for the immunoblotting analysis. Alpha-tubulin was used as an internal control.
- https://cdn.elifesciences.org/articles/85293/elife-85293-fig6-data1-v1.zip
Figure 6—figure supplement 1
Quantitative RT-PCR analysis.
(A) Quantitative RT-PCR analysis of CTPS mRNA abundance in the fat body lysates of 76~80 hour AEL larvae (30 larvae/group; 3 groups/genotype; 2 biological replicates). All data are shown as mean ± …
Figure 6—figure supplement 2
Quantitative RT-PCR analysis.
(A) Quantitative RT-PCR analysis of acc and Fasn1 mRNA abundance in the fat body lysates of 76~80 hour AEL larvae (30 larvae/group; 3 groups/genotype; 2 biological replicates). All data are shown as …
Author response image 1
Quantitative RT-PCR analysis (A) Quantitative RT-PCR analysis of rp49 and GAPDH mRNA abundance in the fat body lysates of the third instar larvae from the indicated genotypes (30 larvae/genotype; 3 groups/genotype, 3 biological replicates).
All data are shown as mean ± S.E.M. ns, no significance, ** P < 0.01, by two-way ANOVA with a Tukey post hoc test.
Author response image 2
Egg hatchability.
(A-C) Egg hatching rate in the indicative lines under HFD condition (n=100 embryos/group, 3 groups/genotype, 2 biological replicates). All data are shown as mean ± S.E.M. ns, no significance by …
Author response image 3
CgG4 in combination with UAS-eGFP indicates fat mass.
In comparison to the wild-type control, the third instar wandering larvae that were alive and expressed eGFP (green) exhibited either Akt or Myc knockdown in the fat body with CgG4 driving. The …
