Comparative proteome analysis of 3T3-L1 adipocyte differentiation using iTRAQ-coupled 2D LC-MS/MS

Feng, Yifei; Zhang, Huoming; Yang, Yanwen; Hu, Huaidong; Sze, Siu Kwan; Wang, Meng; Qian, Jingru; Ren, Hong; Yang, Baolin; Luo, Mingying; Wu, Xiaoqiong; Zhu, Wu; Cai, Wenju; Tong, Jianbin

doi:10.1002/jcb.23223

Cited by 42 publications

(36 citation statements)

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“…The mouse 3T3-L1 pre-adipocytes, which differentiate into mature adipocytes upon hormonal stimulation, were used as the in vitro model of adipogenesis (Green and Meuth, 1974;Mandrup and Lane, 1997;Ye et al, 2011). It is well-known that adipogenic differentiation is controlled by a set of adipogenic transcriptional factors (Farmer, 2006).…”

Section: Resultsmentioning

confidence: 99%

“…The mouse cell line 3T3-L1 was used because its pre-adipocytic state is a well-established in vitro culture model for adipocyte differentiation and its differentiated state is a widely-used in vitro model of mature adipocytes (Green and Kehinde, 1975;Rosen and Spiegelman, 2000;Ye et al, 2011). medium at 37°C, the non-adherent cells were removed by changing the medium, and the rat ADSCs were continuing maintained in basal medium till confluence.…”

Section: Introductionmentioning

confidence: 99%

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Apelin inhibits adipogenesis and lipolysis through distinct molecular pathways

Than

Cheng

Foh

et al. 2012

Molecular and Cellular Endocrinology

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Apelin inhibits adipogenesis and lipolysis through distinct molecular pathways

Than

Cheng

Foh

et al. 2012

Molecular and Cellular Endocrinology

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“…Diet-induced obesity has been shown to differentially regulate protein expression in rat liver, 20 human cerebrospinal fluid, 21 and adipose tissue in culture. 22 Proteomics analysis has identified secretory factors associated with adipogenesis in human adipose stromal vascular fraction cells and differentiated adipocytes. 23 Although proteomic methods have been used to characterize drug transporters and receptors at the BBB, 11,[24][25][26] the changes of BBB proteomes after DIO have not been shown previously.…”

Section: Discussionmentioning

confidence: 99%

Diet-Induced Obesity Suppresses Expression of Many Proteins at the Blood–Brain Barrier

Ouyang

Hsuchou

Kastin

et al. 2013

J Cereb Blood Flow Metab

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The blood-brain barrier (BBB) is a regulatory interface between the central nervous system and the rest of the body. However, BBB changes in obesity and metabolic syndrome have not been fully elucidated. We hypothesized that obesity reduces energy metabolism in the cerebral microvessels composing the BBB, reflected by downregulation of protein expression and function. We performed comparative proteomic analyses in enriched microvessels from the cerebral cortex of mice 2 months after ingestion of a high-fat diet or regular rodent chow. In mice with diet-induced obesity (DIO), there was downregulation of 47 proteins in the cerebral microvessels, including cytoskeletal proteins, chaperons, enzymes, transport-related proteins, and regulators for transcriptional and translational activities. Only two proteins, involved in messenger RNA (mRNA) transport and processing, were upregulated. The changes of these proteins were further validated by quantitative polymerase chain reaction (qPCR), western blotting, and immunofluorescent staining of freshly isolated microvessels, in samples obtained from different batches of mice. The predominant downregulation suggests that DIO suppresses metabolic activity of BBB microvessels. The finding of a hypometabolic state of the BBB in mice at the chronic stage of DIO is unexpected and unprecedented; it may provide novel mechanistic insight into how obesity influences CNS function via regulatory changes of the BBB. Keywords: blood-brain barrier; cerebral microvessels; high-fat diet; mass spectrometry; obesity; proteomics INTRODUCTION Obesity has become one of the most critical problems in human health and is associated with many complications of metabolic diseases. 1,2 Proteomic methods have been used to investigate human and mouse obesity, but most reports have focused on changes in adipose tissue. [3][4][5][6] Recent results suggest that the blood-brain barrier (BBB) is activated in obesity; for example, mice with adult-onset obesity show resistance to leptin transport across the BBB, region-specific astrogliosis, and upregulation of astrocytic leptin receptors. 7,8 The BBB is a regulatory interface between the central nervous system (CNS) and the peripheral circulation. It is a complex threedimensional structure composed of specialized endothelial cells that are reinforced by pericytes, astrocytic endfeet, and extracellular matrix. The tight junctions between the endothelia and the continuous basement membrane prevent diffusion of large non-lipophilic molecules between the CNS parenchyma (brain and spinal cord) and the circulation. The function of the BBB is further reinforced by a high mitochondrial content that ensures sufficient energy, an abundance of enzymes, and metabolically active astrocytes. The functions of the BBB in obesity, however, remain unclear. We predicted that BBB homeostasis is disturbed by metabolic changes in obesity.Although normal transport and secretory functions of the BBB are essential for CNS function, proteomic analyses have shown that BBB activity is ch...

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“…Adipocyte differentiation of 3T3-L1 cells follows the previously established protocol (24,25). Specifically, 2 days after the cells became confluent, adipocyte differentiation was induced (defined as day 0) by incubation with DMEM and 10% FBS, 5 g/ml insulin, 1 M dexamethasone, and 0.5 mM 3-isobutyl-1-methylxanthine.…”

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confidence: 99%