Mice lacking triglyceride synthesis enzymes in adipose tissue are resistant to diet-induced obesity
- Department of Molecular Metabolism, Harvard T.H. Chan School of Public Health, United States
- Department of Cell Biology, Harvard Medical School, United States
- Cell Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, United States
- Broad Institute of Harvard and MIT, United States
- Howard Hughes Medical Institute, United States
Figures
Figure 1 with 3 supplements
ADGAT DKO mice have reduced fat mass and triglycerides in adipose tissue.
(A) Chow-diet-fed ADGAT DKO mice appear normal. Representative photographs of control and ADGAT DKO female mice fed a chow diet. (B) Fat depots were decreased in ADGAT DKO mice. Nuclear magnetic …
Figure 1—figure supplement 1
ADGAT DKO mice have reduced DGAT1 and DGAT2 expression in adipose tissue.
(A) Dgat1 and Dgat2 transcripts levels were decreased in inguinal white adipose tissue (iWAT) and brown adipose tissue (BAT) of ADGAT DKO mice. Relative mRNA levels in iWAT and BAT of male mice fed …
Figure 1—figure supplement 2
ADGAT DKO mice have reduced triglycerides in brown adipose tissue (BAT).
(A) Gross appearance of brown adipose tissue (BAT). (B) H&E-stained sections of inguinal BAT (iBAT) (n=6). Scale bars, 50 µm. (C) Triglycerides and total cholesterol levels in BAT (n=6). (D) [18F]-FD…
Figure 1—figure supplement 3
Most of the triglyceride molecular species were decreased in adipose tissue of ADGAT DKO mice.
(A) Triglycerides were decreased in inguinal white adipose tissue (iWAT) of ADGAT DKO male mice (n=6). Triglyceride levels were measured using Infinity Triglyceride Reagent. (B) Diacylglycerol and …
Figure 2
Adipose tissue triglyceride (TG) stores are required to prevent a torpor-like state during fasting.
(A) Body weights of male mice fed ad libitum or fasted 14 hr (n=10). Ad lib., ad libitum fed. (B) Core body temperature of male mice housed at room temperature and fed ad libitum or fasted for 14 hr …
Figure 3
Lipodystrophy is uncoupled from detrimental metabolic effects in ADGAT DKO mice.
(A) Adiponectin and leptin mRNA levels were moderately increased in inguinal white adipose tissue (iWAT) of ADGAT DKO mice. Relative mRNA levels of leptin and adiponectin in iWAT of chow-diet-fed …
Figure 4 with 1 supplement
ADGAT DKO mice are resistant to diet-induced obesity and glucose intolerance.
(A) ADGAT DKO mice stay lean on a high-fat diet (HFD). Representative photographs of male mice fed on HFD for 12 weeks. (B) Both male and female ADGAT DKO mice gained ~40% less body weight than …
Figure 4—figure supplement 1
ADGAT DKO mice gradually activate an alternative mechanism to synthesize triglycerides.
(A) Lipid droplets (LDs) isolated from inguinal white adipose tissue (iWAT) of mice fed a high-fat diet (HFD). (B) Blood glucose levels in HFD-fed male mice (n=8). (C) Insulin levels in HFD-fed male …
Figure 5 with 3 supplements
Beiging of white adipose tissue (WAT) in ADGAT DKO mice.
(A) ADGAT DKO mice contain multi-locular lipid droplets (LDs) in adipocytes of inguinal WAT (iWAT). H&E-stained sections of iWAT from male mice fed a chow diet and housed at room temperature (n=6). …
Figure 5—figure supplement 1
Analysis of metabolites in inguinal white adipose tissue (iWAT) of ADGAT DKO mice.
(A) Volcano plot showing differentially abundant metabolites in inguinal white adipose tissue (iWAT) of chow-diet-fed male mice (n=8). (B) Volcano plot showing differentially abundant metabolites in …
Figure 5—figure supplement 2
Primary adipocytes cultured from inguinal white adipose tissue (iWAT) of ADGAT DKO mice contain lipid droplets.
(A) Primary adipocytes differentiated from stromal vascular fraction of inguinal white adipose tissue (iWAT) from ADGAT DKO male mice contain lipid droplets (LDs). LDs were stained by BODIPY …
Figure 5—figure supplement 3
Fibroblast growth factor 21 (FGF21) levels were increased in ADGAT DKO mice.
(A) FGF21 transcript levels were increased in inguinal white adipose tissue (iWAT) and livers of ADGAT DKO mice. Relative mRNA levels of FGF21 in iWAT and livers of chow-diet-fed male mice (n=6). (B)…
Videos
Video 1
ADGAT DKO mice enter torpor during fasting.
After 10 hr of fasting ADGAT DKO mice enter a torpor-like state with decreased physical activity and huddling together. Fasting of ADGAT DKO mice also resulted in hypothermia, with body temperatures …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Rabbit polyclonal anti-DGAT1 | In house (Chitraju et al., 2019) | Request Farese and Walther lab for antibodies | |
| Antibody | Rabbit polyclonal anti-DGAT2 | In house (Chitraju et al., 2019) | Request Farese and Walther lab for antibodies | |
| Antibody | Rabbit polyclonal anti-UCP1 | Abcam | Cat# ab10983 | WB (1:1000) |
| Antibody | Total OXPHOS Rodent WB Antibody Cocktail | Abcam | Cat# ab110413 | WB (1:1000) |
| Antibody | Rabbit monoclonal anti-GAPDH | Cell Signaling Technology | Cat# 5174S | WB (1:5000) |
| Antibody | Mouse monoclonal anti-α-Tubulin | Sigma-Aldrich | Cat# T9026 | WB (1:5000) |
| Commercial assay or kit | Power SYBR Green PCR Master Mix | Life Technologies | Cat# 4368706 | |
| Commercial assay or kit | iScript cDNA Synthesis Kit | Bio-Rad | Cat# 170-8891 | |
| Commercial assay or kit | RNeasy Mini Kit | QIAGEN | Cat# 74106 | |
| Commercial assay or kit | QIAzol Lysis Reagent | QIAGEN | Cat# 79306 | |
| Commercial assay or kit | QIAshredder | QIAGEN | Cat# 79656 | |
| Commercial assay or kit | RNase-Free DNase Set | QIAGEN | Cat# 79254 | |
| Commercial assay or kit | SuperSignal West Pico | Thermo Fisher Scientific | Cat# 34580 | |
| Commercial assay or kit | SuperSignal West Femto | Thermo Fisher Scientific | Cat# 34095 | |
| Commercial assay or kit | Infinity Triglycerides Reagent | Thermo Fisher | Cat# TR22421 | |
| Commercial assay or kit | Infinity Cholesterol Reagent | Thermo Fisher | Cat# TR13421 | |
| Commercial assay or kit | Free Glycerol Reagent | Sigma-Aldrich | Cat# F6428 | |
| Commercial assay or kit | HR Series NEFA-HR(2) Color Reagent A | FUJIFILM Medical Systems | Cat# 999-34691 | |
| Commercial assay or kit | HR Series NEFA-HR(2) Solvent A | FUJIFILM Medical Systems | Cat# 995-34791 | |
| Commercial assay or kit | HR Series NEFA-HR(2) Color Reagent B | FUJIFILM Medical Systems | Cat# 991-34891 | |
| Commercial assay or kit | HR Series NEFA-HR(2) Solvent B | FUJIFILM Medical Systems | Cat# 993-35191 | |
| Chemical compound, drug | 1,2-Dioleoyl-rac-glycerol | Sigma-Aldrich | Cat# D8394 | |
| Chemical compound, drug | Oleoyl coenzyme A lithium salt | Sigma-Aldrich | Cat# O1012 | |
| Chemical compound, drug | Oleoyl [14C] Coenzyme A | American Radiolabeled Chemicals, Inc | Cat# ARC 0527 | |
| Chemical compound, drug | Oleic acid [14C] | American Radiolabeled Chemicals, Inc | Cat# ARC 0297 | |
| Chemical compound, drug | Thin Layer Chromatography Plates | Analtech | Cat# P43911 | |
| Chemical compound, drug | Insulin | Lilly Corporation | Humulin R (U 100) | |
| Chemical compound, drug | CL 316,243 hydrate | Sigma-Aldrich | Cat# C5976 | |
| Chemical compound, drug | Glycogen from bovine liver | Sigma-Aldrich | Cat# G0885 | |
| Chemical compound, drug | Protease inhibitors | Roche | Cat# 11873580001 | |
| Other | ADGAT DKO mice | This paper | Request Farese and Walther lab for mice | |
| Other | Dgat1flox/flox mice | (Shih et al., 2009); The Jackson Laboratory | Cat# 017322 | |
| Other | Dgat2flox/flox mice | (Chitraju et al., 2019); The Jackson Laboratory | Cat# 033518 | |
| Other | Adipoq-Cre (Adiponectin-Cre) mice | (Eguchi et al., 2011); The Jackson Laboratory | Cat# 028020 | |
| Other | Chow diet | PicoLab Rodent Diet 20 | Cat# 5053 | |
| Other | Breeder’s diet | PicoLab Rodent Diet 20 | Cat# 5058 | |
| Other | High-fat diet | Envigo | Cat# TD.88137 |
Additional files
-
MDAR checklist
- https://cdn.elifesciences.org/articles/88049/elife-88049-mdarchecklist1-v1.docx
-
Supplementary file 1
List of primers used for quantitative real-time PCR analysis of mouse genes.
- https://cdn.elifesciences.org/articles/88049/elife-88049-supp1-v1.pdf
-
Source data 1
Source data is provided as zip file.
- https://cdn.elifesciences.org/articles/88049/elife-88049-data1-v1.zip
