Journal: Acta Physiologica (Oxford, England)
Article Title: Reversal of Diabesity Through Modulating Sympathetic Inputs to Adipose Tissue Following Carotid Body Resection
doi: 10.1111/apha.70074
Figure Lengend Snippet: Ablation of carotid body (CB) activity through carotid sinus nerve (CSN) resection heightens visceral white adipose tissue (WAT) metabolism in obese dysmetabolic rodents. (A) Panel shows a schematic illustration of the protocol used to evaluate mice WAT oxygen consumption rate (OCR). From (B) to (H) is described the effect of high fat (HF) diet and of CSN resection on the: (B) OCR per minute, reflecting adipose tissue metabolism, before and after stimulation with norepinephrine [15 μM] (left panel) or dopamine [100 nM] (right panel) in mice (3 pieces of tissue from 4–6 animals); (C) average basal OCR in mice (15–27 pieces of tissue from 4–6 animals) before the stimulation with norepinephrine and dopamine; (D) average OCR after stimulation with norepinephrine [15 μM] or dopamine [100 nM] (3 pieces of tissue from 4–6 animals); (E) Illustration of the molecular markers involved in brown adipocytes differentiation as well as the stimuli involved in the beiging of WAT; (F) average expression of PGC1α (92 kDa) ( n = 3–5); (G) average expression of PPARγ (53‐57 kDa) on visceral WAT of rats and mice ( n = 4–5)—representative western blots are shown on the top of the graphs; and (H) percentage of UCP1 protein labeled cells and percentage of mitotrackerTM Red CMXRos labeled cells (top panels) in the perienteric depot ( n = 4–5) in rats and mice; right panels show representative images of UCP1 and MitotrackerTM Red CMXRos labeled cells, Green—UCP1 labeled adipocytes; Red—MitotrackerTM Red CMXRos labeled adipocytes; Blue—DAPI labeled nuclei of the adipocytes; Yellow—Merge of UCP1 and MitotrackerTM Red CMXRos labeled cells. Gray and blue colors represent, respectively, normal chow (NC) and HF rats. Gray and red colors show NC and HF mice, respectively. Den—means animals submitted to CSN denervation/resection. Bars represent mean values ± SEM. Two‐Way ANOVA with Bonferroni multicomparison test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 comparing NC vs. HF groups, # p < 0.05, ## p < 0.01, ### p < 0.001 and ### p < 0.0001 comparing values with and without CSN resection.
Article Snippet: After 1 h of blocking in milk, the membranes were incubated overnight at 4°C with the primary antibodies against β2 receptors (1:200; 47 kDa; Alomone, Jerusalem, Israel), β3 receptors (1:200; 45KDa; Alomone, Jerusalem, Israel), D1R (1:200; 48 kDa; Abcam, Cambridge, UK), D2R (1:200; 49 kDa, Sigma‐Aldrich, Madrid, Spain), Dopamine β hydroxylase (DβH) (1:1000; Merck, Darmstadt, Germany), HSL (1:1000; 83 KDa; Cell Signaling Technology, Massachusetts, EUA), pAMPK (phospho Thr172) (1:1000; 60 kDa; Cell Signaling Technology, Massachusetts, EUA), pATGL (phospho S406) (1:1000; 55 kDa; Abcam, Cambridge, UK), PGC‐1α (1:1000; 92 kDa; Santa Cruz Biotechnology INC, Texas, EUA), PPARγ (1:1000; 53–57 kDa, Cell Signaling Technology, Massachusetts, EUA), TH (1:1000; 60 kDa; Abcam, Cambridge, UK).
Techniques: Activity Assay, Expressing, Western Blot, Labeling
Santa Cruz Biotechnology is a verified supplier 
![Ablation of carotid body (CB) activity through carotid sinus nerve (CSN) resection heightens visceral white adipose tissue (WAT) metabolism in obese dysmetabolic rodents. (A) Panel shows a schematic illustration of the protocol used to evaluate mice WAT oxygen consumption rate (OCR). From (B) to (H) is described the effect of high fat (HF) diet and of CSN resection on the: (B) OCR per minute, reflecting adipose tissue metabolism, before and after stimulation with norepinephrine [15 μM] (left panel) or dopamine [100 nM] (right panel) in mice (3 pieces of tissue from 4–6 animals); (C) average basal OCR in mice (15–27 pieces of tissue from 4–6 animals) before the stimulation with norepinephrine and dopamine; (D) average OCR after stimulation with norepinephrine [15 μM] or dopamine [100 nM] (3 pieces of tissue from 4–6 animals); (E) Illustration of the molecular markers involved in brown adipocytes differentiation as well as the stimuli involved in the beiging of WAT; (F) average expression of PGC1α (92 kDa) ( n = 3–5); (G) average expression of PPARγ (53‐57 kDa) on visceral WAT of rats and mice ( n = 4–5)—representative western blots are shown on the top of the graphs; and (H) percentage of UCP1 protein labeled cells and percentage of mitotrackerTM Red CMXRos labeled cells (top panels) in the perienteric depot ( n = 4–5) in rats and mice; right panels show representative images of UCP1 and MitotrackerTM Red CMXRos labeled cells, Green—UCP1 labeled adipocytes; Red—MitotrackerTM Red CMXRos labeled adipocytes; Blue—DAPI labeled nuclei of the adipocytes; Yellow—Merge of UCP1 and MitotrackerTM Red CMXRos labeled cells. Gray and blue colors represent, respectively, normal chow (NC) and HF rats. Gray and red colors show NC and HF mice, respectively. Den—means animals submitted to CSN denervation/resection. Bars represent mean values ± SEM. Two‐Way ANOVA with Bonferroni multicomparison test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 comparing NC vs. HF groups, # p < 0.05, ## p < 0.01, ### p < 0.001 and ### p < 0.0001 comparing values with and without CSN resection.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6215/pmc12186215/pmc12186215__APHA-241-e70074-g002.jpg)
![Carotid sinus nerve (CSN) resection improves brown adipose tissue (BAT) metabolism in rodents. Effect of high fat (HF) diet and of CSN resection on: (A) Curves of oxygen consumption rate (OCR) per minute, reflecting adipose tissue metabolism, before and after stimulation with norepinephrine [15 μM] (left panel) or dopamine [100 nM] (right panel) in the BAT of mice ( n = 18–27 pieces of tissue from 6–8 animals). (B) Average basal OCR in the BAT in mice ( n = 18–27 pieces of tissue from 6–8 animals) before the stimulation with norepinephrine and dopamine; (C) Average OCR after stimulation with norepinephrine [15 μM] or dopamine [100 nM] ( n = 5–8 animals); (D) percentage of UCP1 protein labeled cells and percentage of mitotrackerTM Red CMXRos labeled cells (top panels) in BAT ( n = 4–5) in rats and mice; bottom panels show representative images of UCP1 and MitotrackerTM Red CMXRos labeled cells, Green—UCP1 labeled adipocytes; Red—MitotrackerTM Red CMXRos labeled adipocytes; Blue—DAPI labeled nuclei of the adipocytes; Yellow—Merge of UCP1 and MitotrackerTM Red CMXRos labeled cells; (E) Rg′ values, reflecting glucose uptake on BAT depots in rats ( n = 5–7); (F) BAT average expression of pATGL (55 kDa) in rats ( n = 4–5); (G) average expression of pAMPK (60 kDa) on BAT of rats and mice ( n = 3–8)—representative western blots are shown on the top of the graphs. Gray and blue colors represent, respectively, normal chow (NC) and HF diet rats. Gray and red colors show NC and HF mice, respectively. Den—means animals submitted to CSN denervation/resection. Bars represent mean values ± SEM. Two‐Way ANOVA with Bonferroni multicomparison test. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 comparing NC vs. HF groups; # p < 0.05, ## p < 0.01, ### p < 0.001 and #### p < 0.0001 comparing values with and without CSN resection.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6215/pmc12186215/pmc12186215__APHA-241-e70074-g001.jpg)
