Using RNA-Seq to Identify Reference Genes of the Transition from Brown to White Adipose Tissue in Goats

Brown adipose tissues have unique non-shivering thermogenesis functions, can be found in newborn ruminate animals, and then are gradually replaced by white adipose tissues in adulthood. For the purpose of exploring the intrinsic mechanism underlying the conversion process from brown (BAT) to white adipose tissue (WAT), it is necessary to utilize Quantitative PCR (qPCR) to study gene expression profiling. In this study, we identified reference genes that were consistently expressed during the transformation from goat BAT to WAT using RNA-seq data. Then, twelve genes were evaluated as candidate reference genes for qPCR in goat perirenal adipose tissue using three tools (geNorm, Normfinder, and BestKeeper). In addition, the selected reference genes were used to normalize the gene expression of PGC-1α and GPAT4. It was found that traditional reference genes, such as GAPDH, RPLP0, HPRT1, and PPIA were not suitable for target gene normalization. In contrast, CTNNB, PFDN5, and EIF3M, selected from RNA sequencing data, showed the least variation and were recommended as the best reference genes during the transformation from BAT to WAT.

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Anti-Obesity Effects of Grateloupia elliptica, a Red Seaweed, in Mice with High-Fat Diet-Induced Obesity via Suppression of Adipogenic Factors in White Adipose Tissue and Increased Thermogenic Factors in Brown Adipose Tissue

Nutrients ◽

10.3390/nu12020308 ◽

2020 ◽

Vol 12 (2) ◽

pp. 308 ◽

Cited By ~ 4

Author(s):

Hyo-Geun Lee ◽

Yu An Lu ◽

Xining Li ◽

Ji-Min Hyun ◽

Hyun-Soo Kim ◽

...

Keyword(s):

Adipose Tissue ◽

Brown Adipose Tissue ◽

White Adipose Tissue ◽

High Fat Diet ◽

Red Seaweed ◽

High Fat ◽

Adipose Tissues ◽

Ppar Γ ◽

Brown Adipose ◽

Treatment Of Obesity

Obesity is a serious metabolic syndrome characterized by high levels of cholesterol, lipids in the blood, and intracellular fat accumulation in adipose tissues. It is known that the suppression of adipogenic protein expression is an effective approach for the treatment of obesity, and regulates fatty acid storage and transportation in adipose tissues. The 60% ethanol extract of Grateloupia elliptica (GEE), a red seaweed from Jeju Island in Korea, was shown to exert anti-adipogenic activity in 3T3-L1 cells and in mice with high-fat diet (HFD)-induced obesity. GEE inhibited intracellular lipid accumulation in 3T3-L1 cells, and significantly reduced expression of adipogenic proteins. In vivo experiments indicated a significant reduction in body weight, as well as white adipose tissue (WAT) weight, including fatty liver, serum triglycerides, total cholesterol, and leptin contents. The expression of the adipogenic proteins, SREBP-1 and PPAR-γ, was significantly decreased by GEE, and the expression of the metabolic regulator protein was increased in WAT. The potential of GEE was shown in WAT, with the downregulation of PPAR-γ and C/EBP-α mRNA; in contrast, in brown adipose tissue (BAT), the thermogenic proteins were increased. Collectively, these research findings suggest the potential of GEE as an effective candidate for the treatment of obesity-related issues via functional foods or pharmaceutical agents.

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Novel Aspects of White Adipose Tissue Browning by Thyroid Hormones

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/a-1020-5354 ◽

2019 ◽

Vol 128 (06/07) ◽

pp. 446-449 ◽

Cited By ~ 4

Author(s):

Kerstin Krause

Keyword(s):

Adipose Tissue ◽

Thyroid Hormones ◽

White Adipose Tissue ◽

Brown Adipocytes ◽

Adipose Tissues ◽

Brown Adipose ◽

Sympathetic Nervous ◽

Tissue Browning ◽

Thermogenic Capacity ◽

Thermogenic Response

AbstractThyroid hormones are essential for the full thermogenic capacity of brown adipose tissue. The thermogenic response of brown adipocytes to thyroid hormones is resulting from the synergistic interaction of thyroid hormones with the sympathetic nervous system. In recent years, evidence has been provided that thyroid hormones also induce the browning of white adipose tissues. This review will provide a brief overview about the recent findings regarding the effects of thyroid hormones on adipose tissue thermogenesis including central and peripheral regulation of white adipose tissue browning.

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Cold-induced lipid dynamics and transcriptional programs in white adipose tissue

BMC Biology ◽

10.1186/s12915-019-0693-x ◽

2019 ◽

Vol 17 (1) ◽

Cited By ~ 9

Author(s):

Ziye Xu ◽

Wenjing You ◽

Yanbing Zhou ◽

Wentao Chen ◽

Yizhen Wang ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Fatty Acid ◽

White Adipose Tissue ◽

Cold Exposure ◽

Fatty Acid Elongation ◽

Rna Seq ◽

Short Term ◽

Lipid Dynamics ◽

Expression Of Genes

Abstract Background In mammals, cold exposure induces browning of white adipose tissue (WAT) and alters WAT gene expression and lipid metabolism to boost adaptive thermogenesis and maintain body temperature. Understanding the lipidomic and transcriptomic profiles of WAT upon cold exposure provides insights into the adaptive changes associated with this process. Results Here, we applied mass spectrometry and RNA sequencing (RNA-seq) to provide a comprehensive resource for describing the lipidomic or transcriptome profiles in cold-induced inguinal WAT (iWAT). We showed that short-term (3-day) cold exposure induces browning of iWAT, increases energy expenditure, and results in loss of body weight and fat mass. Lipidomic analysis shows that short-term cold exposure leads to dramatic changes of the overall composition of lipid classes WAT. Notably, cold exposure induces significant changes in the acyl-chain composition of triacylglycerols (TAGs), as well as the levels of glycerophospholipids and sphingolipids in iWAT. RNA-seq and qPCR analysis suggests that short-term cold exposure alters the expression of genes and pathways involved in fatty acid elongation, and the synthesis of TAGs, sphingolipids, and glycerophospholipids. Furthermore, the cold-induced lipid dynamics and gene expression pathways in iWAT are contrary to those previously observed in metabolic syndrome, neurodegenerative disorders, and aging, suggesting beneficial effects of cold-induced WAT browning on health and lifespan. Conclusion We described the significant alterations in the composition of glyphospholipids, glycerolipids, and sphingolipids and expression of genes involved in thermogenesis, fatty acid elongation, and fatty acid metabolism during the response of iWAT to short-term cold exposure. We also found that some changes in the levels of specific lipid species happening after cold treatment of iWAT are negatively correlated to metabolic diseases, including obesity and T2D.

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Effects of high-carbohydrate diets on lipogenesis in rat interscapular brown adipose tissue

Biochemical Journal ◽

10.1042/bj2060667 ◽

1982 ◽

Vol 206 (3) ◽

pp. 667-669 ◽

Cited By ~ 2

Author(s):

P. John Weaire ◽

Tazeen F. Kanagasabai

Keyword(s):

Adipose Tissue ◽

Brown Adipose Tissue ◽

White Adipose Tissue ◽

White Bread ◽

Lipogenic Enzymes ◽

Adipose Tissues ◽

Interscapular Brown Adipose Tissue ◽

High Carbohydrate ◽

Brown Adipose

Cycloplasmic preparations from brown and white adipose tissues were assayed for three lipogenic enzymes throughout a programme of starvation followed by refeeding on either a normal or a white-bread diet. In the brown adipose tissue of rats fed on a white-bread diet the three enzymes were elevated to levels significantly higher than those in white adipose tissue.

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Photoperiodic regulation of gene expression in brown and white adipose tissue of Siberian hamsters (Phodopus sungorus)

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.2002.282.1.r114 ◽

2002 ◽

Vol 282 (1) ◽

pp. R114-R121 ◽

Cited By ~ 23

Author(s):

Gregory E. Demas ◽

Robert R. Bowers ◽

Timothy J. Bartness ◽

Thomas W. Gettys

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

White Adipose Tissue ◽

Uncoupling Protein ◽

Regulation Of Gene Expression ◽

Ribonuclease Protection Assay ◽

Peroxisome Proliferator ◽

Uncoupling Protein 1 ◽

Siberian Hamsters ◽

Brown Adipose

Siberian hamsters exhibit seasonal fluctuations in white adipose tissue (WAT) mass, with peaks in long “summerlike” days (LDs) and nadirs in short “winterlike” days (SDs). These responses can be mimicked in the laboratory after transfer from LDs to SDs. The purpose of the present study was to test whether changes in WAT and brown adipose tissue (BAT) gene expression that are mediated by the sympathetic nervous system in other obesity models are also associated with seasonal adiposity changes in Siberian hamsters. SDs decreased WAT mass and leptin mRNA, increased WAT β3-adrenoceptor mRNA, and induced retroperitoneal WAT uncoupling protein-1 mRNA (the latter measured by RT-PCR, others measured by ribonuclease protection assay) while increasing BAT uncoupling protein-1 and peroxisome proliferator-activated receptor-γ coactivator-1 mRNAs. These effects were not due to SD-induced gonadal regression and largely occurred before the usual SD-induced decreases in food intake. Thus the SD-induced decreased adiposity of Siberian hamsters may be due to a coordinated suite of WAT and BAT gene transcription changes ultimately increasing lipid mobilization and utilization.

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Adipose tissues of MPC1± mice display altered lipid metabolism-related enzyme expression levels

PeerJ ◽

10.7717/peerj.5799 ◽

2018 ◽

Vol 6 ◽

pp. e5799 ◽

Cited By ~ 4

Author(s):

Shiying Zou ◽

Liye Zhu ◽

Kunlun Huang ◽

Haoshu Luo ◽

Wentao Xu ◽

...

Keyword(s):

Adipose Tissue ◽

Acid Oxidation ◽

Rna Seq ◽

Adipose Tissues ◽

Energy Deficiency ◽

Mitochondrial Pyruvate Carrier ◽

Brown Adipose ◽

Adipocyte Volume ◽

Related Enzyme ◽

Altered Lipid

Mitochondrial pyruvate carrier 1 (MPC1) is a component of the MPC1/MPC2 heterodimer that facilitates the transport of pyruvate into mitochondria. Pyruvate plays a central role in carbohydrate, fatty, and amino acid catabolism. The present study examined epididymal white adipose tissue (eWAT) and intrascapular brown adipose tissue (iBAT) from MPC1± mice following 24 weeks of feeding, which indicated low energy accumulation as evidenced by low body and eWAT weight and adipocyte volume. To characterize molecular changes in energy metabolism, we analyzed the transcriptomes of the adipose tissues using RNA-Sequencing (RNA-Seq). The results showed that the fatty acid oxidation pathway was activated and several genes involved in this pathway were upregulated. Furthermore, qPCR and western blotting indicated that numerous genes and proteins that participate in lipolysis were also upregulated. Based on these findings, we propose that the energy deficiency caused by reduced MPC1 activity can be alleviated by activating the lipolytic pathway.

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Increased expression level of ANGPTL8 in white adipose tissue under acute and chronic cold treatment

Lipids in Health and Disease ◽

10.1186/s12944-021-01547-0 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Hossein Arefanian ◽

Irina Al-Khairi ◽

Nermeen Abu Khalaf ◽

Preethi Cherian ◽

Sina Kavalakatt ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

White Adipose Tissue ◽

Cold Exposure ◽

Uncoupling Protein ◽

Cold Treatment ◽

Control Group ◽

Initial Increase ◽

Mrna Levels ◽

Brown Adipose

Abstract Background Angiopoietin-like proteins (ANGPTL), primarily 3, 4, and 8, play a major role in maintaining energy homeostasis by regulating triglyceride metabolism. This study evaluated the level of ANGPTL3, 4, and 8 in the liver, brown adipose tissue (BAT), and subcutaneous white adipose tissue (SAT) of mice maintained under acute and chronic cold conditions. Methods C57BL/6J mice were exposed to cold temperature (4 °C) for 10 days with food provided ad libitum. Animal tissues were harvested at Day 0 (Control group, n = 5) and Days 1, 3, 5, and 10 (cold treatment groups, n = 10 per group). The expression levels of various genes were measured in the liver, SAT, and BAT. ANGPTL3, 4, and 8 expressions were measured in the liver. ANGPTL4, 8, and genes involved in browning and lipid metabolism [uncoupling protein 1 (UCP1), lipoprotein lipase (LPL), and adipose triglyceride lipase (ATGL)] were measured in SAT and BAT. Western blotting (WB) analysis and immunohistochemistry (IHC) were performed to confirm ANGPTL8 expression in these tissues. Results The expressions of ANGPTL3 and 8 mRNA were significantly reduced in mouse liver tissues after cold treatment (P < 0.05); however, the expression of ANGPTL4 was not significantly altered. In BAT, ANGPTL8 expression was unchanged after cold treatment, whereas ANGPTL4 expression was significantly reduced (P < 0.05). ANGPTL4 levels were also significantly reduced in SAT, whereas ANGPTL8 gene expression exhibited over a 5-fold increase. Similarly, UCP1 gene expression was also significantly increased in SAT. The mRNA levels of LPL and ATGL showed an initial increase followed by a gradual decrease with an increase in the days of cold exposure. ANGPTL8 protein overexpression was further confirmed by WB and IHC. Conclusions This study shows that exposure to acute and chronic cold treatment results in the differential expression of ANGPTL proteins in the liver and adipose tissues (SAT and BAT). The results show a significant reduction in ANGPTL4 in BAT, which is linked to improved thermogenesis in response to acute cold exposure. ANGPTL8 was activated under acute and chronic cold conditions in SAT, suggesting that it is involved in regulating lipolysis and enhancing SAT browning.

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On the characteristics of mitochondrial monoamine oxidase in pancreas and adipose tissues from genetically obese mice

Canadian Journal of Biochemistry ◽

10.1139/o79-024 ◽

1979 ◽

Vol 57 (3) ◽

pp. 197-200 ◽

Cited By ~ 11

Author(s):

J. H. Tong ◽

A. D'Iorio ◽

C. Kandaswami

Keyword(s):

Adipose Tissue ◽

Monoamine Oxidase ◽

White Adipose Tissue ◽

Specific Activity ◽

Obese Mice ◽

Adipose Tissues ◽

Brown Adipose ◽

Mitochondrial Monoamine Oxidase ◽

Mao Activity ◽

Isolated Adipocytes

The substrate specificity of mitochondrial monoamine oxidase (MAO) in pancreatic and adipose tissues of obese mice and their lean counterparts was determined. The pancreatic MAO of obese mice had a greater specific activity than that of the lean mice. The white adipose tissue MAO was found to be more active than the brown adipose MAO in both groups of mice. While there was no appreciable difference in the MAO activities of brown adipose tissues between obese and lean mice, the enzyme from the white adipose tissue of obese mice had a higher specific activity than that of the lean mice. The higher MAO activity in white adipose tissue was observed when tyramine or serotonin was employed as substrate but not with benzylamine. Examination of mitochondrial MAO from epididymal adipocytes revealed marked differences in the properties of the enzyme between whole adipose tissue and isolated adipocytes. The inhibition characteristics of MAO from these tissues were studied with the specific inhibitors clorgyline and deprenyl.

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Adipocyte deletion of HuR induces spontaneous cardiac hypertrophy and fibrosis

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00957.2020 ◽

2021 ◽

Author(s):

Adrienne R. Guarnieri ◽

Sarah R. Anthony ◽

Anamarie Gozdiff ◽

Lisa C. Green ◽

Salma M. Fleifil ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Cardiac Fibrosis ◽

Serum Levels ◽

Tissue Homeostasis ◽

Cardiometabolic Health ◽

Specific Gene ◽

Rna Seq ◽

Tissue Specific ◽

Brown Adipose

Adipose tissue homeostasis plays a central role in cardiovascular physiology, and the presence of thermogenically active brown adipose tissue (BAT) has recently been associated with cardiometabolic health. We have previously shown that adipose tissue-specific deletion of HuR (Adipo-HuR-/-) reduces BAT-mediated adaptive thermogenesis, and the goal of this work was to identify the cardiovascular impacts of Adipo-HuR-/-. We found that Adipo-HuR-/- mice exhibit a hypercontractile phenotype that is accompanied by increased left ventricle wall thickness and hypertrophic gene expression. Furthermore, hearts from Adipo-HuR-/- display increased fibrosis via picrosirius red staining and periostin expression. To identify underlying mechanisms, we applied both RNA-seq and weighted gene co-expression network analysis (WGCNA) across both cardiac and adipose tissue to define HuR-dependent changes in gene expression as well as significant relationships between adipose tissue gene expression and cardiac fibrosis. RNA-seq results demonstrated a significant increase in pro-inflammatory gene expression in both cardiac and subcutaneous white adipose tissue (scWAT) from Adipo-HuR-/- mice that is accompanied by an increase in serum levels of both TNF-ᵯC; and IL-6. In addition to inflammation-related genes, WGCNA identified a significant enrichment in extracellular vesicle-mediated transport and exosome-associated genes in scWAT whose expression most significantly associated with degree of cardiac fibrosis observed in Adipo-HuR-/- mice, implicating these processes as a likely adipose-to-cardiac paracrine mechanism. These results are significant in that they demonstrate the spontaneous onset of cardiovascular pathology in an adipose tissue-specific gene deletion model and contribute to our understanding of how disruptions in adipose tissue homeostasis may mediate cardiovascular disease.

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Acute cold-induced suppression of ob (obese) gene expression in white adipose tissue of mice: mediation by the sympathetic system

Biochemical Journal ◽

10.1042/bj3110729 ◽

1995 ◽

Vol 311 (3) ◽

pp. 729-733 ◽

Cited By ~ 199

Author(s):

P Trayhurn ◽

J S Duncan ◽

D V Rayner

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Lipoprotein Lipase ◽

White Adipose Tissue ◽

Critical Role ◽

Sympathetic System ◽

Obese Gene ◽

Ob Gene ◽

Brown Adipose ◽

White Fat

The effect of acute exposure to cold on the expression of the ob (obese) gene, which encodes a protein that plays a critical role in the regulation of energy balance and body weight, has been examined in epididymal white adipose tissue of mice. Overnight (18 h) exposure of mice to a temperature of 4 degrees C led to the disappearance of ob mRNA in epididymal white fat, and subsequent studies showed that a cold-induced loss of ob mRNA could occur in as little as 2-4 h of exposure to 4 degrees C. When mice exposed to cold for 18 h were returned to the warm (24 degrees C), there was a rapid stimulation of the expression of the ob gene, the mRNA returning within 2.5 h. Administration of noradrenaline led to a reduction in the level of ob mRNA in mice maintained in the warm, while isoprenaline resulted in the disappearance of the mRNA; these changes in ob mRNA were paralleled by similar changes in lipoprotein lipase mRNA. In contrast to white fat, the level of lipoprotein lipase mRNA in brown adipose tissue was increased by noradrenaline and isoprenaline. It is concluded that there is a cold-induced suppression of ob gene expression in white adipose tissue of mice and that this is mediated primarily by the sympathetic system. The profound effect of cold on ob gene expression indicates that the ob system relates to energy expenditure, as well as to satiety.

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