Hypermetabolism in mice carrying a near-complete human chromosome 21
- Department of Physiology, Johns Hopkins University School of Medicine, United States
- Center for Metabolism and Obesity Research, Johns Hopkins University School of Medicine, United States
- Department of Neuroscience, Johns Hopkins University School of Medicine, United States
- Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, United States
- Department of Mental Health, Johns Hopkins Bloomberg School of Public Health, United States
- The Lieber Institute for Brain Development, United States
- Center for Computational Biology, Johns Hopkins University, United States
- Department of Genetic Medicine, Johns Hopkins University School of Medicine, United States
- Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, United States
- Department of Cell Biology, Johns Hopkins University School of Medicine, United States
- Department of Physiology and Cell Biology, The Ohio State University, United States
- Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, United States
- Division of Genome and Cellular Functions, Department of Molecular and Cellular Biology, School of Life Science, Faculty of Medicine, Tottori University, Japan
- Chromosome Engineering Research Center, Tottori University, Japan
Figures
Figure 1
Human chromosome 21 genes are differentially expressed and regulated in mouse adipose tissue, liver, and skeletal muscle.
(A) Graphical representation of human chromosome 21 (Hsa21) and the entire long arm (Hsa21q) region carried by a mouse artificial chromosome in the transchromosomic mouse model (TcMAC21). Four …
Figure 2 with 3 supplements
Hypermetabolism in TcMAC21 mice fed a standard chow.
(A) Body weights of mice fed a standard chow. (B) Body composition analysis of fat and lean mass (relative to body weight). (C) Absolute and relative (normalized to body weight) food intake over a …
Figure 2—figure supplement 1
ELISA data from euploid and MAC21 mice.
Circulating levels of adiponectin (A), leptin (B), growth hormone (C), IGF-1 (D), thyroid hormone (T3, E), and thyroxine (T4, F) in chow-fed mice housed at room temperature (22°C). Circulating …
Figure 2—figure supplement 2
Electron micrograph quantification of pancreatic islet and acinar cell zymogen granules.
(A) Histology of pancreas and quantification of β-islet cross-sectional area (CSA) in TcMAC21 and euploid mice. (B) Pancreatic insulin and somatostatin (SST) contents (normalized to pancreatic …
Figure 2—figure supplement 3
Light and dark cycle body temperature of euploid and MAC21 mice fed a standard chow.
(A) Deep colonic temperature. (B) Interscapular skin temperature. (C) Abdominal skin temperature. (D) Tail skin temperature. Each data point represents the average of 3 days of independent data …
Figure 3 with 1 supplement
TcMAC21 mice are resistant to diet-induced obesity and metabolic dysfunction.
(A) Body weights over time on a high-fat diet and representative mouse images. (B) Body composition analysis of fat and lean mass. (C) Histology of inguinal white adipose tissue (iWAT) and …
Figure 3—figure supplement 1
Representative tissue dissection images of euploid and MAC21 mice fed a high-fat diet.
Representative side-by-side dissection images of (A) heart, (B) kidney, (C) spleen, (D) tibia, (E) inguinal white adipose tissue (iWAT), (F) gonadal white adipose tissue (gWAT), (G) liver, (H) …
Figure 4
Hypermetabolism in TcMAC21 mice fed a high-fat diet (HFD).
(A) Absolute and relative (normalized to body weight) food intake in mice fed an HFD. (B) Fecal energy content, fecal weight, and fecal frequency of TcMAC21 mice and euploid controls. (C) Serum …
Figure 5 with 5 supplements
Hypermetabolism of TcMAC21 mice is uncoupled from changes in adipose and liver transcriptomes.
(A–D) Differentially expressed mouse genes (DEGs), both protein-coding (PCG) and non-protein-coding genes (NPCG) in brown adipose tissue (BAT), liver, gonadal white adipose tissue (gWAT), and …
Figure 5—figure supplement 1
Volcano plot of all qPCR results from brown adipose tissue (BAT), skeletal muscle, and white adipose tissue.
Overlaid results of euploid and MAC21 mice across all conditions assayed (chow diet at 22°C, high-fat diet at 22°C, and high-fat diet at 30°C) for (A) BAT, (B) gastrocnemius (skeletal muscle), (C) …
Figure 5—figure supplement 2
Gene expression analysis in brown adipose tissue.
qPCR analysis of brown adipose tissue genes important for futile cycling, lipid metabolism, mitochondrial function, and thyroid hormone action in euploid and MAC21 male mice fed a standard chow and …
Figure 5—figure supplement 3
Gene expression analysis in inguinal white adipose tissue (iWAT).
qPCR analysis of iWAT genes important for lipid uptake, lipolysis, inflammation, fibrosis, and browning in euploid and MAC21 male mice fed a standard chow diet and housed at 22°C (A), fed a high-fat …
Figure 5—figure supplement 4
Gene expression analysis in gonadal white adipose tissue (gWAT).
qPCR analysis of gWAT genes important for lipid uptake, fatty acid synthesis, lipolysis, inflammation, and fibrosis in euploid and MAC21 male mice fed a standard chow diet and housed at 22°C (A), …
Figure 5—figure supplement 5
Gene expression analysis in liver.
qPCR analysis of liver genes important for lipid uptake, fatty acid synthesis and oxidation, and triglyceride synthesis in euploid and MAC21 male mice fed a standard chow and housed at 22°C (A), fed …
Figure 6 with 9 supplements
Sarcolipin overexpression in skeletal muscle drives TcMAC21 hypermetabolism.
(A) RNA-sequencing analysis reveals changes in TcMAC21 relative to euploid skeletal muscle (gastrocnemius). Volcano plot of skeletal muscle transcriptome (transcripts from the mouse genome only). …
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Figure 6—source data 1
Top left—Original uncropped membrane from imager showing all channels (red/green/blue), with the sarcolipin protein band labeled as SLN appearing blue.
Top right—Original uncropped membrane showing a black and white image of the blue channel only. SLN band is marked with an arrow. The dotted outline represents where the membrane image was cropped for use in Figure 6. Bottom left—Original uncropped membrane from imager showing all channels (red/green/blue), with the GAPDH protein band labeled and appearing blue. Bottom right—Original uncropped membrane showing a black and white image of the blue channel only. GAPDH band is marked with an arrow. The dotted outline represents where the membrane image was cropped for use in Figure 6.
- https://cdn.elifesciences.org/articles/86023/elife-86023-fig6-data1-v1.zip
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Figure 6—source data 2
Top left—Original uncropped membrane from imager showing all channels (red/green/blue), with the OXPHOS complex protein bands labeled as CI, CII, CIII, CIV, and CV appearing blue.
Top right—Original uncropped membranes of different exposures showing black and white images of the blue channel only. OXHPHOS complex protein bands are labeled. The dotted outline represents where the membrane image was cropped for use in Figure 6. Bottom left—Original uncropped membrane from imager showing all channels (red/green/blue), with the GAPDH protein band labeled and appearing blue. Bottom right—Original uncropped membrane showing a black and white image of the blue channel only. GAPDH band is marked with an arrow. The dotted outline represents where the membrane image was cropped for use in Figure 6.
- https://cdn.elifesciences.org/articles/86023/elife-86023-fig6-data2-v1.zip
Figure 6—figure supplement 1
The top up- and downregulated genes in skeletal muscle based on RNA-sequencing data.
(A) Top 100 upregulated protein-coding genes. (B) Top 100 downregulated protein-coding genes. (C) All upregulated non-protein-coding genes. (D) All downregulated non-protein-coding genes. n=5 …
Figure 6—figure supplement 2
Gene expression analysis in skeletal muscle.
qPCR analysis of gastrocnemius skeletal muscle genes important for general metabolism, fast- and slow-twitch fiber types, futile cycling, thyroid hormone action, and calcium handling in euploid and …
Figure 6—figure supplement 3
Skeletal muscle cross-sectional area (CSA) analysis of MAC21 and euploid mice fed a high-fat diet.
(A–B) Representative wheat germ agglutinin (WGA)-stained gastrocnemius skeletal muscle samples. (C) Muscle fiber CSA analysis of TcMAC21 mice (n=7) and euploid controls (n=8). Mice were housed at …
Figure 6—figure supplement 4
Mitochondrial respirometry analysis of gastrocnemius and quadricep muscle.
(A and C) Average group gastrocnemius (Gastroc) oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), …
Figure 6—figure supplement 5
Mitochondrial respirometry analysis of soleus and plantaris muscle.
(A and C) Average group soleus oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), respectively. (B and D) …
Figure 6—figure supplement 6
Mitochondrial respirometry analysis of extensor digitorum longus (EDL) and brown adipose tissue (BAT).
(A and C) Average group EDL oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), respectively. (B and D) …
Figure 6—figure supplement 7
Mitochondrial respirometry analysis of liver and tongue.
(A and C) Average group liver oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), respectively. (B and D) …
Figure 6—figure supplement 8
Mitochondrial respirometry analysis of heart.
(A and C) Average group heart oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), respectively. (B and D) …
Figure 6—figure supplement 9
Mitochondrial respirometry analysis of inguinal (iWAT) and gonadal (gWAT) white adipose tissue.
(A and C) Average group iWAT oxygen consumption rate (OCR) traces using NADH as a substrate for euploid (Eu) and MAC21 male mice fed a standard chow or high-fat diet (HFD), respectively. (B and D) …
Figure 7 with 1 supplement
Increased energy expenditure (EE) persists in TcMAC21 mice housed at thermoneutrality.
(A) Body weights of TcMAC21 mice and euploid controls housed at thermoneutrality (30°C) for 8 weeks. (B) Body composition analysis of absolute and relative (normalized to body weight) fat and lean …
Figure 7—figure supplement 1
Histology of high-fat diet (HFD)-fed euploid and MAC21 mice housed at thermoneutrality (30°C).
Representative hematoxylin and eosin (H&E)-stained sections of inguinal white adipose tissue (iWAT, A–B), gonadal white adipose tissue (gWAT, D and E), brown adipose tissue (BAT, H and I), liver (J …
Figure 8
Potential regulators of sarcolipin expression in skeletal muscle.
(A) Volcano plot of skeletal muscle transcriptome (transcripts from the mouse genome only). The lower dotted line denotes significance at the p-value cut-off (p=0.05), and the upper dotted line …
Author response image 1
Tables
Author response table 1
| Food intake (kcal/day) | |||
|---|---|---|---|
| Chow-fed | Euploid | TcMAC21 | p-value |
| Day 1 | 12.93±1.75 | 13.39±1.07 | 0.52 |
| Day 2 | 11.95±2.41 | 13.16±1.47 | 0.23 |
| Day 3 | 12.20±2.60 | 13.32±1.68 | 0.3 |
| Day 4 | 12.74±2.56 | 13.41±2.26 | 0.57 |
| Day 5 (fasted) | 0 | 0 | |
| Day 6 (Refed) | 17.55±2.97 | 17.32±1.46 | 0.83 |
| Food intake (kcal/day) | |||
| HFD-fed | Euploid | TcMAC21 | p-value |
| Day 1 | 15.52±3.68 | 13.00±1.71 | 0.06 |
| Day 2 | 12.30±5.6 | 12.0±2.22 | 0.8 |
| Day 3 | 14.67±5.90 | 13.80±2.60 | 0.69 |
| Day 4 | 15.28±3.40 | 16.15±2.48 | 0.55 |
| Day 5 (fasted) | 0 | 0 | |
| Day 6 (Refed) | 13.04±2.46 | 17.74±2.30 | 0.001 |
Additional files
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Supplementary file 1
Tissue weights of chow-fed euploid (n=7) and MAC21 (n=6) male mice at termination of study.
Eu, euploid; BW, body weight; gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp1-v1.xlsx
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Supplementary file 2
Indirect calorimetry analysis of male (16.5 weeks of age) euploid (n=8) and MAC21 (n=9) mice fed a standard chow.
Eu, euploid; VO2, rate of oxygen consumption; VCO2, rate of carbon dioxide production; RER, respiratory exchange ratio; EE, energy expenditure.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp2-v1.xlsx
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Supplementary file 3
Tissue weight of high-fat diet-fed male mice (33 weeks of age; fed diet for 16.5 weeks) at termination of the study.
Eu, euploid; gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue; BAT, brown adipose tissue.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp3-v1.xlsx
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Supplementary file 4
Complete blood count of euploid (n=5) and MAC21 (n=6) male mice fed a high-fat diet (50 weeks of age; on diet for 13 weeks).
RBC, red blood cell count; HGB, hemoglobin; HCT, hematocrit; MCV, mean corpuscular (erythrocyte) volume; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; RET, reticulocyte count; PLT, platelet count; WBC, white blood cells count; NEUT, neutrophil count; LYMPH, lymphocyte count; MONO, monocyte count; EO, eosinophil count; BASO, basophil count; Eu, euploid.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp4-v1.xlsx
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Supplementary file 5
Indirect calorimetry analysis of male (25 weeks of age) euploid (n=8) and TcMAC21 (n=9) mice fed a high-fat diet (8.5 weeks on diet).
Eu, euploid; VOC, rate of oxygen consumption; VCO2, rate of carbon dioxide production; RER, respiratory exchange ratio; EE, energy expenditure.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp5-v1.xlsx
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Supplementary file 6
List of all differentially expressed genes (DEGs) in the gonadal white adipose tissue (gWAT) of euploid and MAC21 male mice fed a high-fat diet housed at 22°C.
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp6-v1.xlsx
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Supplementary file 7
List of all differentially expressed genes (DEGs) in the inguinal white adipose tissue (iWAT) of euploid and MAC21 male mice fed a high-fat diet housed at 22°C.
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp7-v1.xlsx
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Supplementary file 8
List of all differentially expressed genes (DEGs) in the liver of euploid and MAC21 male mice fed a high-fat diet housed at 22°C.
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp8-v1.xlsx
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Supplementary file 9
List of all differentially expressed genes (DEGs) in the brown adipose tissue (BAT) of euploid and MAC21 male mice fed a high-fat diet housed at 22°C.
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp9-v1.xlsx
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Supplementary file 10
List of all differentially expressed genes (DEGs) in the skeletal muscle (gastrocnemius) of euploid and MAC21 male mice fed a high-fat diet housed at 22°C.
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp10-v1.xlsx
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Supplementary file 11
RNA-sequence data from skeletal muscle (gastrocnemius) showing expression values for atrophy-related genes, contractile and structural genes, fibrosis and injury repair-related genes, and muscle wasting-related genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp11-v1.xlsx
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Supplementary file 12
Indirect calorimetry analysis in thermoneutral conditions (31°C) after 2-week acclimatization period of male (55 weeks of age) euploid (n=5) and MAC21 (n=4) mice fed a high-fat diet (18 weeks on diet).
Eu, euploid; VO2, rate of oxygen consumption; VCO2, rate of carbon dioxide production; RER, respiratory exchange ratio; EE, energy expenditure.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp12-v1.xlsx
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Supplementary file 13
Tissue weights of male MAC21 (n=4) and euploid (n=5) mice (55 weeks of age) housed at thermoneutrality (31°C) and fed a high-fat diet (HFD, 18 weeks on diet).
Eu, euploid; gWAT, gonadal white adipose tissue; iWAT, inguinal white adipose tissue; BAT, brown adipose tissue.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp13-v1.xlsx
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Supplementary file 14
List of all differentially expressed genes (DEGs) in the skeletal muscle (gastrocnemius) of euploid and MAC21 male mice fed a high-fat diet housed at thermoneutrality (31°C).
Lists include: detected human genes and all significantly up- and downregulated mouse genes.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp14-v1.xlsx
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Supplementary file 15
List of primers used for all qPCR analysis of gene expression.
- https://cdn.elifesciences.org/articles/86023/elife-86023-supp15-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/86023/elife-86023-mdarchecklist1-v1.docx
