Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: Phenotypic characterization of GDF15 knockout mice on a high fat diet (HFD) . (A and B) Body weight and percent body weight gain of wild type (WT) and GDF15 KO mice fed a 60% HFD; Inset, final body weight and percent body weight gain. (C) Weight of epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver and brown adipose tissue (BAT), harvested at the end of the study, 25 weeks of HFD (n = 13,9). (D) Weight of total hepatic lipids in g; total lipid extracted from 25 mg tissue was normalized to total liver weight (n = 14,9). (E) Lipid droplet area (Percent liver area) determined from histological analyses of haematoxylin/eosin (H&E) stained liver sections (n = 9,8). (F) Plasma triglycerides (TG), cholesterol, leptin, alanine transaminase (ALT) and aspartate transaminase (AST) from random fed mice after 16 weeks of HFD-feeding (n = 12,11). (G) Blood glucose, plasma insulin and HOMA-IR levels from 6 h fasted mice, after 16 weeks of HFD feeding (n = 16–29). (H and I) Blood glucose levels during intraperitoneal (ip) glucose tolerance test (GTT) and percent change from initial blood glucose levels during insulin tolerance test (ITT) after 16 weeks of HFD feeding. Inset, area under the curve analysis of glucose over time. (J) Plasma FGF21 levels from random fed mice at 16 weeks of HFD feeding (n = 7,9). (K) FGF21 mRNA expression in tissues from WT and GDF15 KO mice after 25 weeks HFD feeding (n = 8–11). All data are means ± S.D ∗/∗∗/∗∗∗/∗∗∗∗ - p < 0.05/0.01/0.001/0.0001.

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Knock-Out, Staining, Clinical Proteomics, Expressing

Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: GDF15 mRNA expression within high fat diet fed mouse and human adipose tissue . (A) In situ hybridization analysis of GDF15 mRNA (red) from 18-week-old high fat diet (HFD) fed wild type (WT) mouse epididymal adipose tissue. Black arrows indicate GDF15 staining in foamy macrophages contributing to the formation of ‘crown-like structures’. Blue arrow indicates GDF15 staining within adipocytes. (B) Representative haematoxylin/eosin image of epididymal tissue from 18-week old HFD fed WT mouse. Black arrows indicate infiltrating cells. (C) Representative image (corresponding to the image in B) of epididymal tissue from 18-week-old HFD fed WT mouse stained with the macrophage marker F4/80 confirming that the cells contributing to the ‘crown like structures’ are macrophages. (D and E) GDF15 and EMR1 mRNA expression from 14 week old Chow or HFD fed WT mouse epididymal tissue fractionated into adipocytes, CD11b negative (−) and CD11b positive (+) stromal vascular fractions (SVF) (n = 3–4). (F) Correlation of GDF15 expression with EMR1 expression in epididymal tissue from 14 week Chow or HFD fed wild type mice (n = 8). (G–I) Human subcutaneous adipose tissue GDF15 gene expression levels in the TwinsUK adipose study associated with estimated macrophage proportion in adipose tissue (G), and macrophage markers CD68 and EMR1 (H and I). Each point represents data from a single individual. Plotted gene expression residuals of GDF15, CD68 and EMR1 were adjusted for age, BMI and RNA-Seq technical covariates. (J and K) GDF15 gene expression levels in the ‘Obese study’ associated with macrophage marker CD68 in human subcutaneous and visceral adipose tissue. (L and M) In situ hybridization analysis of Gdf15 mRNA (red) from 18-week old HFD fed WT mouse liver and brown adipose tissue. (N) FGF21 mRNA expression from 14 week Chow or HFD fed WT mouse epididymal tissue fractionated in to adipocytes, CD11b negative (−) and CD11b positive (+) stromal vascular fraction (SVF) (n = 3–4).

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Expressing, In Situ Hybridization, Staining, Marker, Gene Expression, RNA Sequencing

Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: Phenotypic characterization of LysM-Cre mediated GDF15 macrophage knockout mouse on a high fat diet (HFD) . (A) GDF15 mRNA expression from 24-week-old high fat diet (HFD) fed wild type (WT) and LysM-GDF15 KO mouse epididymal adipose tissue fractionated into adipocytes and CD11b positive (+) stromal vascular fraction (SVF) (n = 4). (B) GDF15 mRNA expression in tissues from 24 week HFD fed WT and LysM-GDF15 KO mice. Epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), brown adipose tissue (BAT), liver and kidney (n = 4–7). (C) Plasma GDF15 levels from random fed mice at indicated weeks on HFD (n = 15–18). (D) FGF21 mRNA expression in tissues from 24-week HFD fed WT and LysM-GDF15 KO mice (n = 4–7). (E) Plasma FGF21 levels from random fed mice at indicated weeks on HFD (n = 11–18). (F and G) Body weight and percent body weight gain of WT and LysM-GDF15 KO mice fed a 60% HFD; Inset, final body weight and percent body weight gain. (H) Weight of epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver and brown adipose tissue (BAT) harvested at the end of the study, 24 weeks of HFD (n = 16–18). (I) Weight of total hepatic lipids in g; total lipid extracted from 25 mg tissue was normalized to total liver weight (n = 16,17). (J) Plasma leptin levels in mice from random fed mice after 16 weeks HFD-feeding (n = 16–18). (K) Blood glucose, plasma insulin and HOMA-IR levels from 6 h fasted mice, after 16 weeks of HFD feeding (n = 16,18). (L and M) Blood glucose levels during ip glucose tolerance test (GTT) and percent change from initial blood glucose levels during insulin tolerance test (ITT) after 16 weeks of HFD feeding. Inset, area under the curve analysis of glucose over time. (N) Plasma triglycerides (TG), cholesterol, alanine transaminase (ALT) and aspartate transaminase (AST) from random fed mice, after 16 weeks of HFD feeding (n = 9–11). All data are means ± S.D ∗/∗∗ - p < 0.05/0.01.

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Knock-Out, Expressing, Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: Phenotypic characterization of bone marrow deleted GDF15 knockout mouse on a high fat diet . (A) GDF15 mRNA expression in tissues from 24-week old high fat diet (HFD) fed WT and BMT-GDF15 KO mice. Epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT) and brown adipose tissue (BAT) (n = 8–12). (B) Plasma GDF15 levels from random fed mice at the onset (week 0) and after 12 weeks of HFD feeding (n = 12). (C) FGF21 mRNA expression from indicated tissues of 24 week HFD fed WT and BMT-GDF15 KO mice (n = 8–12). (D) Plasma FGF21 levels from random fed mice at the onset (week 0) and after 12 weeks of HFD feeding (n = 12). (E and F) Body weight and percent body weight gain of WT and BMT-GDF15 KO mice fed a 60% HFD; Inset, final body weight and percent body weight gain. (G) Weight of epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver and brown adipose tissue (BAT) harvested at the end of the study, 24 weeks of HFD (n = 3–12). (H) Plasma triglycerides (TG), cholesterol, leptin, alanine transaminase (ALT) and aspartate transaminase (AST) from random fed mice, after 16 weeks of HFD feeding (n = 12). (I) Weight of total hepatic lipids in g; total lipid extracted from 25 mg tissue was normalized to total liver weight (n = 12). (J) Blood glucose, plasma insulin and HOMA-IR levels from 6 h fasted mice after 16 weeks of HFD feeding (n = 7,6). (K and L) Blood glucose levels during ip glucose tolerance test (GTT) and percent change from initial blood glucose levels during insulin tolerance test (ITT) after 16 weeks of HFD feeding. Inset, area under the curve analysis of glucose over time. All data are means ± S.D ∗/∗∗/∗∗∗/∗∗∗∗ - p < 0.05/0.01/0.001/0.0001.

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Knock-Out, Expressing, Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: Phenotypic characterization of Alb-Cre mediated GDF15 hepatocyte knockout mouse on a high fat diet . (A) GDF15 mRNA expression in tissues from 24-week old high fat diet (HFD) fed wild type (WT) and Alb-GDF15 KO mice. Epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver, brown adipose tissue (BAT) and kidney (n = 10–16). (B) Plasma GDF15 levels from random fed mice at the onset (week 0) and after 12 weeks of HFD feeding (n = 15,13). (C) FGF21 mRNA expression in indicated tissues from 24-week HFD fed WT and Alb-GDF15 KO mice (n = 10–13). (D) Plasma FGF21 levels from random fed mice at the onset (week 0) and after 12 weeks of HFD feeding (n = 15,13). (E and F) Body weight and percent body weight gain of WT and Alb-GDF15 KO mice fed a 60% HFD; Inset, final body weight and percent body weight gain. (G) Weight of epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver and brown adipose tissue (BAT) harvested at the end of the study, 24 weeks of HFD (n = 15,11). (H) Plasma leptin, triglycerides (TG), cholesterol, alanine transaminase (ALT) and aspartate transaminase (AST) from random fed mice, after 16 weeks of HFD feeding (n = 15,13). (I) Weight of total hepatic lipids in g; total lipid extracted from 25 mg tissue was normalized to total liver weight (n = 15,16). (J and K) Blood glucose levels during ip glucose tolerance test (GTT) and percent change from initial blood glucose levels during insulin tolerance test (ITT) after 16 weeks of HFD feeding. Inset, area under the curve analysis of glucose over time. (L) Blood glucose, plasma insulin and HOMA-IR levels from 6 h fasted mice, after 16 weeks of HFD feeding (n = 14,13). All data are means ± S.D ∗/∗∗/∗∗∗/∗∗∗∗ - p < 0.05/0.01/0.001/0.0001.

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Knock-Out, Expressing, Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Combined genetic deletion of GDF15 and FGF21 has modest effects on body weight, hepatic steatosis and insulin resistance in high fat fed mice

doi: 10.1016/j.molmet.2022.101589

Figure Lengend Snippet: Phenotypic characterization of dKO knockout mice on a high fat diet . (A) Liver GDF15 and (B) FGF21 mRNA expression from 24-week-old high fat diet (HFD) fed WT, FGF21 KO, GDF15 KO and FGF21/GDF15 double knockout (dKO) mice (n = 5). (C and D) Plasma GDF15 and FGF21 from random fed mice at indicated time after high fat feeding (n = 3–11). (E and F) Body weight and percent body weight gain of WT, FGF21 KO, GDF15 KO and dKO mice fed a 60% HFD; Inset, final body weight and percent body weight gain. (G and H) Blood glucose levels during ip glucose tolerance test (GTT) and percent change from initial blood glucose levels during insulin tolerance test (ITT) after 16 weeks of HFD feeding. Inset, area under the curve analysis of glucose over time. (I) Blood glucose, plasma insulin and HOMA-IR levels from 6 h fasted mice after 16 weeks of HFD feeding (n = 19–29). (J) Weight of epididymal white adipose tissue (eWAT), subcutaneous white adipose tissue (scWAT), liver and brown adipose tissue (BAT) harvested at the end of the study, 24 weeks of HFD (n = 11–17). (K) Weight of total hepatic lipids; total lipid extracted from 25 mg tissue was normalized to total liver weight (n = 9–17). (L) Lipid droplet area (Percent liver area) determined from histological analyses of H&E stained liver sections (n = 8–11). (M) Plasma triglycerides (TG) from random fed mice, after 24 weeks of HFD feeding (n = 8). (N) Hepatic mRNA expression of genes involved in lipid metabolism (n = 9–17). All data are means ± S.D ∗/∗∗/∗∗∗/∗∗∗∗ - p < 0.05/0.01/0.001/0.0001. Please note that the data for WT and GDF15KO overlaps with the data presented in Figure 1 .

Article Snippet: Briefly, one-cell stage embryos (obtained from super-ovulated wild type C57Bl/6N females fertilised in vitro with sperm from homozygous GDF15 Tm1a male) were injected into the pronucleus with 100ng/ul StemMACS Flp Recombinase mRNA (Miltenyi Biotec) then transferred into the uteri of pseudo pregnant recipient females (F1 hybrids from C57Bl/6J female × CBA/Ca male crosses).

Techniques: Knock-Out, Expressing, Double Knockout, Clinical Proteomics, Staining