High-Throughput High-Content Functional Image Analysis of Neuronal Proteins Implicated in Parkinson’s Disease

Blaas, Eva; Kesteren, Ronald E. van

doi:10.1007/978-1-61779-111-6_16

Cited by 1 publication

(1 citation statement)

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“…Stained cells were analyzed by Cellomics Arrayscan high-content microscopy (Thermo Scientific, The Netherlands) at 10× magnification. From each well, 80 image fields were scanned and analyzed using the compartmental analysis bio application (v3.0), designed to quantify compartmentalized changes in fluorescence . End points measured were total number of cells and adipocytes, adipocyte cell size, and the number of fat droplets per cell.…”

Section: Methodsmentioning

confidence: 99%

Transcriptional and Epigenetic Mechanisms Underlying Enhanced in Vitro Adipocyte Differentiation by the Brominated Flame Retardant BDE-47

Kamstra

Hrubá

Blumberg

et al. 2014

Environ. Sci. Technol.

112

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Recent studies suggest that exposure to endocrine-disrupting compounds (EDCs) may play a role in the development of obesity. EDCs such as the flame retardant 2,2′,4,4′-tetrabrominated diphenyl ether (BDE-47) have been shown to enhance adipocyte differentiation in the murine 3T3-L1 model. The mechanisms by which EDCs direct preadipocytes to form adipocytes are poorly understood. Here, we examined transcriptional and epigenetic mechanisms underlying the induction of in vitro adipocyte differentiation by BDE-47. Quantitative high content microscopy revealed concentration-dependent enhanced adipocyte differentiation following exposure to BDE-47 or the antidiabetic drug troglitazone (TROG). BDE-47 modestly activated the key adipogenic transcription factor peroxisome proliferator-activated receptor gamma (PPARγ) in COS7 cells, transiently transfected with a GAL4 reporter construct. Increased gene expression was observed for Pparγ2, leptin (Lep), and glucose-6-phophatase catalytic subunit (G6pc) in differentiated 3T3-L1 cells after BDE-47 exposure compared to TROG. Methylation-sensitive high resolution melting (MS-HRM) revealed significant demethylation of three CpG sites in the Pparγ2 promoter after exposure to both BDE-47 and TROG in differentiated 3T3-L1 cells. This study shows the potential of BDE-47 to induce adipocyte differentiation through various mechanisms that include Pparγ2 gene induction and promoter demethylation accompanied by activation of PPARγ, and possible disruption of glucose homeostasis and IGF1 signaling.

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Section: Methodsmentioning

confidence: 99%