Increased adipose tissue lymphatic vessel density inhibits thermogenesis through elevated neurotensin levels

Figures

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  FIGURE 1. Overexpression of VEGF-D expands adipose lymphatic density but does not impact overall adipose tissue physiology. (A): Immunofluorescence of lymphatic (green; LYVE-1) and blood (red; Endomucin) in brown adipose tissue (BAT) of Control and AdipoVD mice. (B) Immunofluorescence of lymphatic (green; LYVE-1) and blood (red; Endomucin) in subcutaneous white adipose tissue (SQAT) of Control and AdipoVD mice. (C) Body composition quantification of Control and AdipoVD mice by magnetic resonance. (D) Serum glucose levels during an oral glucose tolerance test in Control and AdipoVD mice. (E) Average area under the curve (AUC) calculated from glucose levels of oral glucose tolerance test. (F) Browning and thermogenesis-associated gene expression in AdipoVD BAT relative to Controls. (G) Browning and thermogenesis-associated gene expression in AdipoVD SQAT relative to Controls. (A–E) n = 10 control/n = 7 AdipoVD. (F–G); n = 11 for control and AdipoVD. Blue = DAPI. Bars = 100 µm.

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  FIGURE 2. Lymphangiogenesis in AdipoVD mice result in increased neurotensin production. (A) Neurotensin and neurotensin receptor relative mRNA expression in BAT of AdipoVD mice compared to Controls. (B) Neurotensin and neurotensin receptor relative mRNA expression in SQAT of AdipoVD mice compared to Controls. (C) ELISA quantification of neurotensin concentrations from BAT normalized to tissue protein levels. (D) ELISA quantification of neurotensin concentrations from SQAT normalized to tissue protein levels. (E) Immunofluorescence of neurotensin (green; Nts) and lymphatics (red; LYVE-1) in the BAT depot from Control and AdipoVD mice on HFD. (A–E) n = 10 control/n = 7 AdipoVD. Bars = 50 µm.

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  FIGURE 3. Temperature dysfunction in AdipoVD mice as a result of increased neurotensin levels. (A) Body temperature of Control and AdipoVD mice during acute under cold exposure at 4°C. (B) Browning and thermogenesis-associated gene expression in AdipoVD BAT relative to Controls under acute cold exposure. (C) Immunofluorescence of potential thermogenesis (green; UCP-1) and lymphatic (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (C) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (D) Browning and thermogenesis-associated gene expression in AdipoVD SQAT relative to Controls following cold exposure. (E) Immunofluorescence of potential thermogenesis (green; UCP-1) and lymphatic (red; LYVE-1) from SQAT depot of Control and AdipoVD following cold exposure. (A–E) n = 10 control/n = 7 AdipoVD. Blue = DAPI. Bars = 100 µm.

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  FIGURE 4. Inhibition of neurotensin activity ameliorate temperature dysfunction in AdipoVD mice. (A) Body temperature of Control and AdipoVD mice during acute under cold exposure at 4°C after neurotensin receptor inhibition. (B) Browning and thermogenesis-associated gene expression in AdipoVD BAT relative to Controls under acute cold exposure after neurotensin receptor inhibition. (C) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (D) Browning and thermogenesis-associated gene expression in AdipoVD SQAT relative to Controls following cold exposure and neurotensin receptor inhibition. (E) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from SQAT depot of Control and AdipoVD following cold exposure and neurotensin receptor inhibition. (A–E) n = 10 control/n = 7 AdipoVD. Blue = DAPI. Bars = 100 µm.