scholarly journals Development of an Optimized Clearing Protocol to Examine Adipocyte Subpopulations in White Adipose Tissue

2021 ◽  
Vol 4 (2) ◽  
pp. 39
Author(s):  
Quyen Vi Luong ◽  
Andreas Israel ◽  
Rita Sharma ◽  
Siegfried Ussar ◽  
Kevin Y. Lee

Organic solvent dibenzyl ether (DBE)-based protocols have been widely used in adipose tissue clearing. However, benzyl alcohol/benzyl benzoate (BABB)-based clearing has been shown to offer better transparency in other tissues. The addition of diphenyl ether (DPE) to BABB (BABB-D4) is often included to preserve fluorescent signals, but its effects on adipose tissue transparency and shrinkage have not been explored. Distinct adipocyte subpopulations contribute to its cellular composition and biological activity. Here, we compared clearing solvents to create an optimized clearing methodology for the study of adipocyte subpopulations. Adipose tissues were cleared with BABB, BABB-D4, and DBE, and post-clearing transparency and tissue shrinkage were measured. An optimized protocol, including BABB-D4 clearing, delipidation, and extensive immunofluorescence blocking steps, was created to examine the spatial distribution of Wt-1 positive progenitor-derived (Type-1) adipocytes in intact mesenteric fat. Both BABB and BABB-D4 lead to significantly increased tissue transparency with reduced tissue shrinkage compared to DBE-cleared adipose tissue. Type-1 adipocytes are found in a clustered distribution with predominant residence in fat associated with the ileum and colon. This paper details an optimized clearing methodology for adipose tissue with increased tissue transparency and reduced shrinkage, and therefore will be a useful tool for investigating adipose tissue biology.

2019 ◽  
Vol 51 (10) ◽  
pp. 671-677 ◽  
Author(s):  
Maurício Martins da Silva ◽  
Carlos Frederico Lima Gonçalves ◽  
Leandro Miranda-Alves ◽  
Rodrigo Soares Fortunato ◽  
Denise P. Carvalho ◽  
...  

AbstractPlastics are ubiquitously present in our daily life and some components of plastics are endocrine-disrupting chemicals, such as bisphenol A and phthalates. Herein, we aimed to evaluate the effect of plastic endocrine disruptors on type 1 and type 2 deiodinase activities, enzymes responsible for the conversion of the pro-hormone T4 into the biologically active thyroid hormone T3, both in vitro and in vivo. Initially, we incubated rat liver type 1 deiodinase and brown adipose tissue type 2 deiodinase samples with 0.5 mM of the plasticizers, and the deiodinase activity was measured. Among them, only BPA was capable to inhibit both type 1 and type 2 deiodinases. Then, adult male Wistar rats were treated orally with bisphenol A (40 mg/kg b.w.) for 15 days and hepatic type 1 deiodinase and brown adipose tissue type 2 deiodinase activities and serum thyroid hormone concentrations were measured. In vivo bisphenol A treatment significantly reduced hepatic type 1 deiodinase activity but did not affect brown adipose tissue type 2 deiodinase activity. Serum T4 levels were higher in bisphenol A group, while T3 remained unchanged. T3/T4 ratio was decreased in rats treated with bisphenol A, reinforcing the idea that peripheral metabolism of thyroid hormone was affected by bisphenol A exposure. Therefore, our results suggest that bisphenol A can affect the metabolism of thyroid hormone thus disrupting thyroid signaling.


2002 ◽  
Vol 88 (08) ◽  
pp. 345-353 ◽  
Author(s):  
Erik Maquoi ◽  
Diego Demeulemeester ◽  
Berthe Van Hoef ◽  
Désiré Collen ◽  
H. Lijnen

SummaryA nutritionally induced obesity model was used to investigate the modulation of fibrinolytic and gelatinolytic activity during the development of adipose tissue.Five week old male mice were fed a standard fat diet (SFD, 13% kcal as fat) or a high fat diet (HFD, 42% kcal as fat) for up to 15 weeks. The HFD resulted in body weights of 31 ± 0.9 g, 38 ± 2.0 g and 47 ± 1.9 g at 5, 10 and 15 weeks, respectively; corresponding values for mice on the SFD were 26 ± 0.6 g, 31 ± 0.9 g and 31 ± 1.2 g (all p < 0.001). The weight of the isolated subcutaneous (SC) or gonadal (GON) fat after 15 weeks of HFD was 1,870 ± 180 mg or 1,470 ± 160 mg, as compared to 250 ± 58 mg or 350 ± 71 mg for the SFD (p < 0.001). The HFD induced marked time-dependent hyperglycemia and elevated levels of triglycerides and total cholesterol. The HFD diet also induced a marked hypertrophy of the adipocytes as compared to the SFD, e.g. diameter of 83 ± 3.0 µm versus 52 ± 4.2 µm for GON adipocytes at 15 weeks (p < 0.005). Plasma plasminogen activator inhibitor-1 (PAI-1) levels were higher in mice on the HFD as compared to the SFD; they were comparable in extracts of SC or GON adipose tissue, whereas at different time points tissue-type (t-PA) and urokinase-type (u-PA) plasminogen activator activity was somewhat lower in the adipose tissues of mice on HFD. Gelatinolytic activity (mainly MMP-2) was detected in SC but not in GON adipose tissue of mice on SFD, and decreased on the HFD. In situ zymography on cryosections did not reveal different fibrinolytic activities in SC or GON adipose tissues of the HFD as compared to the SFD groups, whereas significantly lower gelatinolytic and higher caseinolytic activities were detected in SC and GON tissues of mice on the HFD (p ≤ 0.05). The fibrillar collagen content was lower in adipose tissue of mice on HFD. Thus, in this model time-dependent development of adipose tissue appears to be associated with modulation of proteolytic activity.


Endocrinology ◽  
2011 ◽  
Vol 152 (5) ◽  
pp. 1789-1799 ◽  
Author(s):  
Shiho Fujisaka ◽  
Isao Usui ◽  
Yukiko Kanatani ◽  
Masashi Ikutani ◽  
Ichiro Takasaki ◽  
...  

Diet-induced obesity is reported to induce a phenotypic switch in adipose tissue macrophages from an antiinflammatory M2 state to a proinflammatory M1 state. Telmisartan, an angiotensin II type 1 receptor blocker and a peroxisome proliferator-activated receptor-γ agonist, reportedly has more beneficial effects on insulin sensitivity than other angiotensin II type 1 receptor blockers. In this study, we studied the effects of telmisartan on the adipose tissue macrophage phenotype in high-fat-fed mice. Telmisartan was administered for 5 wk to high-fat-fed C57BL/6 mice. Insulin sensitivity, macrophage infiltration, and the gene expressions of M1 and M2 markers in visceral adipose tissues were then examined. An insulin- or a glucose-tolerance test showed that telmisartan treatment improved insulin resistance, decreasing the body weight gain, visceral fat weight, and adipocyte size without affecting the amount of energy intake. Telmisartan reduced the mRNA expression of CD11c and TNF-α, M1 macrophage markers, and significantly increased the expressions of M2 markers, such as CD163, CD209, and macrophage galactose N-acetyl-galactosamine specific lectin (Mgl2), in a quantitative RT-PCR analysis. A flow cytometry analysis showed that telmisartan decreased the number of M1 macrophages in visceral adipose tissues. In conclusion, telmisartan improves insulin sensitivity and modulates adipose tissue macrophage polarization to an antiinflammatory M2 state in high-fat-fed mice.


2012 ◽  
Vol 44 (11) ◽  
pp. 797-803 ◽  
Author(s):  
D. Ignacio ◽  
R. Fortunato ◽  
R.A. Neto ◽  
D. da Silva Silvestre ◽  
M. Nigro ◽  
...  

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 212-212
Author(s):  
Marina Miquilini ◽  
Nicole R Hardy ◽  
Kellie Enger ◽  
Phyllis Dieter ◽  
Alejandro E Relling ◽  
...  

Abstract Estradiol administration increases mammary parenchyma growth and can increase the mass of the mammary fat pad (MFP) in calves. Estradiol can affect fatty acid metabolism, but it is unknown how estradiol affects the fatty acid profile of the MFP and if these effects are consistent across adipose stores. The objective of this investigation was to determine if fatty acid profiles were affected by estradiol administration and if this response is adipose tissue type specific. Holstein heifer calves were reared on a common diet and administered 12 daily injections prior to euthanasia at 82 days of age. Injections were either daily injections of corn oil (n = 4; CON), 9 injections of corn oil followed by 3 injections of estradiol (n = 4; SHORT), or 12 injections of estradiol (n = 4; LONG). Fatty acids were extracted from MFP and subcutaneous adipose tissues samples and analyzed using gas chromatography. Data were analyzed using a mixed model considering the effect of treatment, adipose tissue depot, their interaction, and the random effect of animal. Only significant treatment effects and treatment-adipose type interactions are presented here (Table 1). Estradiol administration (SHORT and LONG) increased total C18:1 fatty acids (P = 0.05), and tended to increase non-C18:2 t10 c12 CLAs, C18:3, and total omega-3 fatty acids (P ≤ 0.1) in the MFP and subcutaneous adipose tissues relative to CON. There was a treatment by tissue interaction for C18:1 trans-10 (P = 0.01), and a tendency for C18:1 trans-11 and C18:2 trans-10 cis-12 (P ≤ 0.1) indicating that estradiol’s effects on fatty acids are tissue specific. Together, these results indicate that the MFP responds differently to estradiol than subcutaneous adipose tissues and that these alterations are associated with different periods of induced mammary growth via estradiol.


Antioxidants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1090
Author(s):  
Ursula Abou-Rjeileh ◽  
G. Andres Contreras

Lipid mobilization in adipose tissues, which includes lipogenesis and lipolysis, is a paramount process in regulating systemic energy metabolism. Reactive oxygen and nitrogen species (ROS and RNS) are byproducts of cellular metabolism that exert signaling functions in several cellular processes, including lipolysis and lipogenesis. During lipolysis, the adipose tissue generates ROS and RNS and thus requires a robust antioxidant response to maintain tight regulation of redox signaling. This review will discuss the production of ROS and RNS within the adipose tissue, their role in regulating lipolysis and lipogenesis, and the implications of antioxidants on lipid mobilization.


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