Synthesis of LipidGreen2 and its application in lipid and fatty liver imaging

Chun, Hang-Suk; Jeon, Jong Hyun; Pagire, Haushabhau S.; Lee, Jae Hong; Chung, Hee-Chung; Park, Myoung Joo; So, Juhoon; Ryu, Jae-Ho; Kim, Cheol‐Hee; Ahn, Jin Hee; Bae, Myung Ae

doi:10.1039/c3mb70022d

Cited by 24 publications

(23 citation statements)

References 26 publications

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“…Zebrafish is a vertebrate model that has been widely used to study lipid biology including lipid metabolism and the genes regulating lipid processing [ 29 ]. We have recently synthesized LipidGreen2 that selectively stains fat deposits in live zebrafish [ 30 , 31 ], which may support the relevance of use of this zebrafish-based LipidGreen2 staining in evaluating the tanshinone IIA’s lipid-reducing effect. To see its efficacy in vivo, we next investigated whether tanshinone IIA inhibits lipid accumulation in zebrafish by LipidGreen2 staining.…”

Section: Resultsmentioning

confidence: 99%

Anti-Adipogenic Effects on 3T3-L1 Cells and Zebrafish by Tanshinone IIA

Park

Obiang-Obounou

Lee

et al. 2017

IJMS

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Tanshinone IIA is a diterpene quinone isolated from the roots of Salvia miltiorrhiza bunge that has traditionally been used in China for the treatment of cardiovascular and cerebrovascular disorders. Although there is recent evidence showing that tanshinone IIA has an anti-obesity effect, its underlying mechanism of anti-obesity effect is poorly understood. Here, we investigated the effect of tanshinone IIA on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Notably, tanshinone IIA at 10 μM concentration greatly reduced lipid accumulation and triglyceride (TG) contents during 3T3-L1 preadipocyte differentiation, suggesting its anti-adipogenic effect. On mechanistic levels, tanshinone IIA reduced the expression levels of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and perilipin A but also the phosphorylation levels of signal transducer and activator of transcription-3/5 (STAT-3/5) in differentiating 3T3-L1 cells. In addition, tanshinone IIA strongly inhibited leptin and resistin mRNA expression in differentiating 3T3-L1 cells. Importantly, the tanshinone IIA’s lipid-reducing effect was also seen in zebrafish. In sum, these findings demonstrate that tanshinone IIA has anti-adipogenic effects on 3T3-L1 cells and zebrafish, and its anti-adipogenic effect on 3T3-L1 cells is largely attributable to the reduced expression and/or phosphorylation levels of C/EBP-α, PPAR-γ, FAS, perilipin A, and STAT-3/5.

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Section: Resultsmentioning

confidence: 99%

Anti-Adipogenic Effects on 3T3-L1 Cells and Zebrafish by Tanshinone IIA

Park

Obiang-Obounou

Lee

et al. 2017

IJMS

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“…Furthermore, Nile red and BODIPY were stained the lysosome-related organelles rather than the major C. elegans fat storage compartment. In the previous studies on LipidGreen1 and its derivatives, LipidGreen1 was capable of a brighter and less non-specific staining for the detection of neutral lipids and fat deposits compared to the commercially available Nile red and BODIPY ® 493/503 (Lee et al 2011 ; Chun et al 2013 ). Therefore, we attempted to examine a possibility for the more reliable detection of bacterial PHB, one of the promising biodegradable plastics, using the lipophilic fluorescent LipidGreen1.…”

Section: Discussionmentioning

confidence: 99%

“…LipidGreen1 is a new small fluorescence probe with an indolin-3-one skeleton, which successfully stained lipid droplets in 3T3-L1 and HepG2 cells and fat deposits in zebrafish (Chun et al 2013 ; Lee et al 2011 ). LipidGreen1 could be used to detecting bacterial polyesters including PHB.…”

Section: Introductionmentioning

confidence: 99%

A lipophilic fluorescent LipidGreen1-based quantification method for high-throughput screening analysis of intracellular poly-3-hydroxybutyrate

Choi

Yang³

et al. 2015

AMB Expr

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Poly-3-hydroxybutyrate (PHB), the most abundant type of polyhydroxyalkanoates (PHA) is synthesized inside a variety of microorganisms as a primary candidate for industrial PHB production. Lipophilic dyes such as Nile red and BODIPY have been used to quantify intracellular PHB, but their uses have often been limited in terms of sensitivity and accuracy. In this study, a newly developed lipophilic fluorescent dye LipidGreen1 was used to quantify intracellular PHB. LipidGreen1 stained viable colonies by adding the dye into the medium which enabled the effective selection of PHB-positive cells. Furthermore, the fluorescence intensity of LipidGreen1 maintained its fluorescence intensity much longer than that of Nile red. The fluorescence intensities of intracellular PHB stained by LipidGreen1 accurately agreed with PHB contents measured by gas chromatography. In addition, internalization of LipidGreen1 in Escherichia coli cell was not necessary to obtain quantitative measurements. PHB-synthase mutants were differentiated by fluorescence intensities with a good correlation to increased levels of PHB production. These results show that LipidGreen1 is sensitive and accurate in high-throughput screening of newly isolated and genetically modified bacteria with enhanced PHB production.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-015-0131-6) contains supplementary material, which is available to authorized users.

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“…Larvae were fasted for 12 h before staining in order to empty the digestive tract, as the presence of food in the gut may give a high background signal, thereby making it difficult to visualize adipocytes. The protocol for LipidGreen staining was performed as described in Chun et al, 2013 [55]. LipidGreen solution with a final concentration of 1 µM was prepared in E3 medium.…”

Section: Methodsmentioning

confidence: 99%

Altered Adipogenesis in Zebrafish Larvae Following High Fat Diet and Chemical Exposure Is Visualised by Stimulated Raman Scattering Microscopy

Broeder

Moester

Kamstra

et al. 2017

IJMS

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Early life stage exposure to environmental chemicals may play a role in obesity by altering adipogenesis; however, robust in vivo methods to quantify these effects are lacking. The goal of this study was to analyze the effects of developmental exposure to chemicals on adipogenesis in the zebrafish (Danio rerio). We used label-free Stimulated Raman Scattering (SRS) microscopy for the first time to image zebrafish adipogenesis at 15 days post fertilization (dpf) and compared standard feed conditions (StF) to a high fat diet (HFD) or high glucose diet (HGD). We also exposed zebrafish embryos to a non-toxic concentration of tributyltin (TBT, 1 nM) or Tris(1,3-dichloroisopropyl)phosphate (TDCiPP, 0.5 µM) from 0–6 dpf and reared larvae to 15 dpf under StF. Potential molecular mechanisms of altered adipogenesis were examined by qPCR. Diet-dependent modulation of adipogenesis was observed, with HFD resulting in a threefold increase in larvae with adipocytes, compared to StF and HGD. Developmental exposure to TBT but not TDCiPP significantly increased adipocyte differentiation. The expression of adipogenic genes such as pparda, lxr and lepa was altered in response to HFD or chemicals. This study shows that SRS microscopy can be successfully applied to zebrafish to visualize and quantify adipogenesis, and is a powerful approach for identifying obesogenic chemicals in vivo.

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Synthesis of LipidGreen2 and its application in lipid and fatty liver imaging

Cited by 24 publications

References 26 publications

Anti-Adipogenic Effects on 3T3-L1 Cells and Zebrafish by Tanshinone IIA

Anti-Adipogenic Effects on 3T3-L1 Cells and Zebrafish by Tanshinone IIA

A lipophilic fluorescent LipidGreen1-based quantification method for high-throughput screening analysis of intracellular poly-3-hydroxybutyrate

Altered Adipogenesis in Zebrafish Larvae Following High Fat Diet and Chemical Exposure Is Visualised by Stimulated Raman Scattering Microscopy

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