scholarly journals Embryonic cardiospecific knockout of α-E-catenin gene leads to alteration of energy metabolism in adult heart

2017 ◽  
Vol 23 (2) ◽  
pp. 65-69
Author(s):  
V. Balatskyy ◽  
L. Macewicz ◽  
O. Piven
Keyword(s):  
Lipid Metabolism ◽  
Energy Metabolism ◽  
Transgenic Mice ◽  
Lipid Droplets ◽  
Metabolic Disorders ◽  
Conditional Knockout ◽  
Heart Hypertrophy ◽  
Oil Red O Staining ◽  
Oil Red O

Previously we have shown that the α-E-catenin knockout in the embryonic heart leads to hypertrophy in adult and activation of canonical Wntsignaling. Heart hypertrophy is also accompanied by metabolic disorders, but role of the α-E-catenin in these processes is not known. Aim of our work is to study the effect of α-E-catenin deletion on the lipid metabolism in the heart. Methods. In our experiment we have used α-Е-catenin conditional knockout and αMHC-Cre transgenic mice. We have utilized histological (Oil Red O staining) and molecular biological (Western blot) methods. Results. α-Е-catenin deletion leads to accumulation of lipid droplets in myocardium, and to violation of expression and phosphorylation of key regulators of lipid metabolism (Ampk, Pparα, Acc, Hsl). Conclusions. Ous results suggest that α-Е-catenin deletion leads to inhibition of lipid metabolism in the heart.

scholarly journals Type III Collagen is Required for Adipogenesis and Actin Stress Fibre Formation in 3T3-L1 Preadipocytes

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 156
Author(s):  
Mohammad Al Hasan ◽  
Patricia E. Martin ◽  
Xinhua Shu ◽  
Steven Patterson ◽  
Chris Bartholomew
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Type Iii Collagen ◽  
Type Iii ◽  
Actin Stress Fibre ◽  
Matrix Gene ◽  
Gene Transcripts ◽  
Polymerase Chain ◽  
Oil Red O Staining ◽  
Oil Red O

GPR56 is required for the adipogenesis of preadipocytes, and the role of one of its ligands, type III collagen (ColIII), was investigated here. ColIII expression was examined by reverse transcription quantitative polymerase chain reaction, immunoblotting and immunostaining, and its function investigated by knockdown and genome editing in 3T3-L1 cells. Adipogenesis was assessed by oil red O staining of neutral cell lipids and production of established marker and regulator proteins. siRNA-mediated knockdown significantly reduced Col3a1 transcripts, ColIII protein and lipid accumulation in 3T3-L1 differentiating cells. Col3a1−/− 3T3-L1 genome-edited cell lines abolished adipogenesis, demonstrated by a dramatic reduction in adipogenic moderators: Pparγ2 (88%) and C/ebpα (96%) as well as markers aP2 (93%) and oil red O staining (80%). Col3a1−/− 3T3-L1 cells displayed reduced cell adhesion, sustained active β-catenin and deregulation of fibronectin (Fn) and collagen (Col4a1, Col6a1) extracellular matrix gene transcripts. Col3a1−/− 3T3-L1 cells also had dramatically reduced actin stress fibres. We conclude that ColIII is required for 3T3-L1 preadipocyte adipogenesis as well as the formation of actin stress fibres. The phenotype of Col3a1−/− 3T3-L1 cells is very similar to that of Gpr56−/− 3T3-L1 cells, suggesting a functional relationship between ColIII and Gpr56 in preadipocytes.

193 Single Cell RNA-sequencing Reveals a Role of Lipid Metabolism in Muscle Satellite Cells

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 104-105
Author(s):  
Shihuan Kuang ◽  
Feng Yue ◽  
Stephanie Oprescu
Keyword(s):  
Gene Expression ◽  
Lipid Metabolism ◽  
Cell Fate ◽  
Satellite Cell ◽  
Satellite Cells ◽  
Lipid Droplets ◽  
Self Renewal ◽  
Droplet Dynamics ◽  
Single Cell Rna Sequencing

Abstract Single Cell RNA-sequencing (scRNA-seq) is a powerful technique to deconvolute gene expression of various subset of cells intermingled within a complex tissue, such as the skeletal muscle. We first used scRNA-seq to understand dynamics of cell populations and their gene expression during muscle regeneration in murine limb muscles. This leads to the identification of a subset of satellite cells (the resident stem cells of skeletal muscles) with immune gene signatures in regenerating muscles. Next, we used scRNA-seq to examine gene expression dynamics of satellite cells at various status: quiescence, activation, proliferation, differentiation and self-renewal. This analysis uncovers stage-dependent changes in expression of genes related to lipid metabolism. Further analyses lead to the discovery of previously unappreciated dynamics of lipid droplets in satellite cells; and demonstrate that the abundance of the lipid droplets in newly divided satellite daughter cells is linked to cell fate segregation into differentiation versus self-renewal. Perturbation of lipid droplet dynamics through blocking lipolysis disrupts cell fate homeostasis and impairs muscle regeneration. Finally, we show that lipid metabolism regulates the function of satellite cells through two mechanisms. On one hand, lipid metabolism functions as an energy source through fatty acid oxidation (FAO), and blockage of FAO reduces energy production that is critical for satellite cell function. On the other hand, lipid metabolism generates bioactive molecules that influence signaling transduction and gene expression. In this scenario, lipid metabolism and FAO regulate the intracellular levels of acetyl-coA and selective acetylation of PAX7, a pivotal transcriptional factor underlying function of satellite cells. These results together reveal for the first time a critical role of lipid metabolism and lipid droplet dynamics in muscle satellite cell fate determination and regenerative function; and underscore a potential role of dietary fatty acids in satellite cell-dependent muscle development, growth and regeneration.

scholarly journals ANTIHYPERGLYCEMIC ACTIVITY OF TINOCRISPOSIDE BY STIMULATING 3T3-L1 ADIPOCYTE CELL DIFFERENTIATION

2018 ◽  
Vol 11 (11) ◽  
pp. 494
Author(s):  
Adek Zamrud Adnan ◽  
Muhammad Taher ◽  
Annisa Fauzana ◽  
Tika Afriani ◽  
Dewi Imelda Roesma ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Lipid Droplets ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Antihyperglycemic Activity ◽  
Significant Difference ◽  
Adipocyte Cell ◽  
Oil Red O Staining ◽  
Oil Red O

Objective: The aim of this study is to investigate the antihyperglycemic activity of tinocrisposide by stimulating 3T3-L1 adipocyte cell differentiation. Tinocrisposide is a furanoditerpene glycoside that was isolated from Tinospora crispa (Menispermaceae).Methods: Adipocyte cell differentiation activity of tinocrisposide in interval concentrations of 50, 25, 12.5, and 6.25 μg/ml has been investigated on 3T3-L1 cell line using insulin of 1 μg/ml as a positive and Dulbecco’s modified Eagle media (DMEM) as a negative control group. The effect of tinocrisposide was quantified with oil red O staining method by measuring an absorbance of lipid solution in isopropanol at a wavelength (λ) of 520 nm.Results: Tinocrisposide in the concentrations of 50, 25, 12.5, and 6.25 μg/ml insulin of 1 μg/ml and DMEM groups showed absorbance value of 0.7669, 0.7253, 0.6563, 0.6481, 0.954, and 0.2653, respectively. It was found that there was a significant difference statistically in lipid droplets accumulation among all groups (p<0.05) and tinocrisposide at a concentration of 50 μg/ml showed the highest lipid droplets accumulation in 3T3-L1 adipocyte cells.Conclusion: From the study, it could be concluded that tinocrisposide was able to stimulate the differentiation of adipocyte cell and had antihyperglycemic activity.

scholarly journals Exon 9-deleted CETP inhibits full length-CETP synthesis and promotes cellular triglyceride storage

2020 ◽  
Vol 61 (3) ◽  
pp. 422-431 ◽  
Author(s):  
Lahoucine Izem ◽  
Yan Liu ◽  
Richard E. Morton
Keyword(s):  
Lipid Metabolism ◽  
Lipid Droplets ◽  
Full Length ◽  
Intracellular Lipid ◽  
Transfer Protein ◽  
Lipid Phenotype ◽  
Exon 9 ◽  
And Storage

Cholesteryl ester transfer protein (CETP) exists as full-length (FL) and exon 9 (E9)-deleted isoforms. The function of E9-deleted CETP is poorly understood. Here, we investigated the role of E9-deleted CETP in regulating the secretion of FL-CETP by cells and explored its possible role in intracellular lipid metabolism. CETP overexpression in cells that naturally express CETP confirmed that E9-deleted CETP is not secreted, and showed that cellular FL- and E9-deleted CETP form an isolatable complex. Coexpression of CETP isoforms lowered cellular levels of both proteins and impaired FL-CETP secretion. These effects were due to reduced synthesis of both isoforms; however, the predominate consequence of FL- and E9-deleted CETP coexpression is impaired FL-CETP synthesis. We reported previously that reducing both CETP isoforms or overexpressing FL-CETP impairs cellular triglyceride (TG) storage. To investigate this further, E9-deleted CETP was expressed in SW872 cells that naturally synthesize CETP and in mouse 3T3-L1 cells that do not. E9-deleted CETP overexpression stimulated SW872 triglyceride synthesis and increased stored TG 2-fold. Expression of E9-deleted CETP in mouse 3T3-L1 cells produced a similar lipid phenotype. In vitro, FL-CETP promotes the transfer of TG from ER-enriched membranes to lipid droplets. E9-deleted CETP also promoted this transfer, although less effectively, and it inhibited the transfer driven by FL-CETP. We conclude that FL- and E9-deleted CETP isoforms interact to mutually decrease their intracellular levels and impair FL-CETP secretion by reducing CETP biosynthesis. E9-deleted CETP, like FL-CETP, alters cellular TG metabolism and storage but in a contrary manner.

1 PATHWAYS AND CELLULAR FUNCTIONS INFLUENCED BY INSULIN TREATMENT DURING OOCYTE MATURATION – A TRANSCRIPTOME STUDY OF IN VITRO-PRODUCED BOVINE DAY 8 BLASTOCYSTS

2015 ◽  
Vol 27 (1) ◽  
pp. 93 ◽  
Author(s):  
D. Laskowski ◽  
Y. Sjunnesson ◽  
R. Båge ◽  
M. A. Sirard ◽  
H. Gustafsson ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Cycle ◽  
Lipid Metabolism ◽  
Energy Metabolism ◽  
Metabolic Disorders ◽  
Insulin Treatment ◽  
Oxidative Stress Response ◽  
Cellular Growth ◽  
Cellular Functions

Insulin as a key metabolic hormone has crucial functions in metabolic regulation in all mammals. Deviation of its physiological concentration occurs in metabolic disorders as obesity and diabetes in humans or negative energy balance and overfeeding in the cow. As these metabolic disorders are strongly correlated with reproductive disturbances, we investigated the effect of insulin during oocyte maturation on gene expression of bovine Day 8 blastocysts (BC8) by transcriptome analysis. Abattoir-derived oocytes (n = 882) were divided into 3 groups and in vitro matured for 22 h by adding insulin (H: High 10 µg mL–1; L: Low 0.1 µg mL–1 and Z: Zero, control). This was followed by standard in vitro production (IVP) and evaluation of developmental rates up to blastocyst stage. BC8 (n = 120) were pooled in groups of 10 and total RNA was extracted by parallel gDNA and total RNA-extraction (AllPrepDNA/RNA micro kit, cat no. 80284, Qiagen®, Valencia, CA, USA) for analyses of the transcriptome. All samples (4 biological replicates/group) resulted in RIN-values >7.5. RNA amplification, cDNA synthesis, purification, and labelling were performed and 825 ng of Cy3- and Cy5-labelled linearly amplified aRNA was hybridized on the Agilent-manufactured EmbryoGENE-slides in a 2-colour dye swap design. An empirical Bayes moderated t-test was applied to search for the differentially expressed transcripts (DET) between control and insulin-treated groups, using the ‘limma’ package in R (www.r-project.org). The DET were defined as having a 1.5-fold change difference between treatment and control and P < 0.05. Pathways and molecular functions influenced by insulin treatment were analysed by using Ingenuity Pathway Analysis (IPA; Ingenuity® Systems, www.ingenuity.com). As a global pattern, insulin treatment induced an up-regulation of genes. In total, 202 DET in the H and 142 DET in the L group were found where 104 DET were common in both insulin groups. Fifteen selected candidate genes chosen for qPCR validation and 12 (80%) showed similar expression patterns as the microarray data. DET relevant for following cellular functions were found in H: Cell Cycle, Cellular Compromise, Lipid Metabolism, Molecular Transport, Small Molecule Biochemistry respective L: Cell Morphology, Cellular Growth and Proliferation, Cell Cycle, Carbohydrate Metabolism and Cellular Assembly and Organization. The top canonical pathways influenced were Epithelial Adherens Junction Signalling and Remodelling, Germ Cell Sertoli Cell Junction Signalling and NRF2-mediated Oxidative Stress Response. Correlatively, blastocyst rates on Day 8 were significantly lower in H and L v. Z (P < 0.05). The transcriptome data could explain the mechanisms behind the impaired development, as genes involved in cellular growth and energy metabolism in Day 8 blastocysts were affected. The fact that transcripts related to NRF2-mediated oxidative stress response and lipid metabolism are up-regulated suggests that insulin induces dysregulation of cellular functions and energy metabolism leading to impaired embryo developmental potential.Funded by FORMAS.

scholarly journals Acylated Ghrelin and The Regulation of Lipid Metabolism in The Intestine

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
N. Auclair ◽  
N. Patey ◽  
L. Melbouci ◽  
Y. Ou ◽  
L. Magri-Tomaz ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Lipid Droplets ◽  
Physiological Saline ◽  
Fatty Acid Uptake ◽  
Intimate Relationship ◽  
Acid Uptake ◽  
Acylated Ghrelin ◽  
Adipose Tissues ◽  
Syrian Golden Hamsters

AbstractAcylated ghrelin (AG) is a gastrointestinal (GI) peptide mainly secreted by the stomach that promotes cytosolic lipid droplets (CLD) hypertrophy in adipose tissues and liver. However, the role of AG in the regulation of lipid metabolism in the intestine remains unexplored. This study aimed at determining whether AG influences CLD production and chylomicron (CM) secretion in the intestine. The effects of AG and oleic acid on CLD accumulation and CM secretion were first investigated in cultured Caco-2/15 enterocytes. Intestinal lipid metabolism was also studied in Syrian Golden Hamsters submitted to conventional (CD) or Western (WD) diets for 8 weeks and continuously administered with AG or physiological saline for the ultimate 2 weeks. In cultured Caco-2/15 enterocytes, CLD accumulation influenced CM secretion while AG reduced fatty acid uptake. In WD hamsters, continuous AG treatment amplified chylomicron output while reducing postprandial CLD accumulation in the intestine. The present study supports the intimate relationship between CLD accumulation and CM secretion in the intestine and it underlines the importance of further characterizing the mechanisms through which AG exerts its effects on lipid metabolism in the intestine.

scholarly journals Korean Traditional Fermented Soybean Paste (Doenjang) Regulate Renin-Angiotensin System (RAS) in 3T3-L1 Adipocytes

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 791-791
Author(s):  
Hayoung Woo ◽  
Jung Eun Park ◽  
Youn-Soo Cha
Keyword(s):  
Blood Pressure ◽  
Lipid Accumulation ◽  
Lipid Droplets ◽  
Renin Angiotensin System ◽  
Angiotensin System ◽  
Renin Angiotensin ◽  
Fermented Soybean ◽  
Soybean Paste ◽  
Oil Red O Staining ◽  
Oil Red O

Abstract Objectives Doenjang, the Korean traditional fermented soybean paste, contains much salt. There is a concern that cardiovascular disease may occur due to such high salinity. Nevertheless, previous studies have demonstrated functional properties of doenjang anti-obesity and anti-cancer effects. Furthermore, in our recent studies, we showed that the anti-hypertensive effect of doenjang through renin-angiotensin system (RAS) regulation. Doenjang regulated the RAS to improve lipid metabolism in adipose tissue, which had a positive effect on blood pressure control. Therefore, we expected to find the exact mechanism of action or target point of doenjang in adipocyte using 3T3-L1 cells. Methods In this study, 3T3-L1 cells were treated with doenjang and RAS blockers, Losartan (10−4 M), and Captopril (10−4 M), were treated as positive control which suppresses AT1R and ACE, respectively. Non-cytotoxic concentrations of samples were selected as per MTT assay and added with induction media, harvested after 4 days for RNA extraction. Lipid droplets were detected by Oil Red O staining. Results Doenjang downregulated mRNA levels of peroxisome proliferator-activated receptor-γ (Pparg), RAS related genes such as angiotensinogen (Agt), Renin (Ren), and aldosterone-releasing factors (P &lt; 0.05). Especially, angiotensin convert enzyme (Ace) and angiotensin II receptor 2 (Agtr2) levels were decreased by doenjang treatment. Doenjang reduced the lipid accumulation, which was confirmed from the Oil Red O staining of lipid droplets. As a result, it is revealed that doenjang not only inhibits lipid accumulation in adipocytes but also may inhibit ACE in 3T3-L1 adipocytes through a mechanism similar to the effect of Captopril. Conclusions These data are consistent with our animal study. It have been shown to regulate blood pressure through lipid improvement and ACE inhibition despite high salt content in doenjang. Funding Sources This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MEST) (No. 2018R1A2B6006477).

Secreted-Osteopontin Contributes to Brown Adipogenesis In Vitro via a CD44-Dependent Pathway

2019 ◽  
Vol 51 (11) ◽  
pp. 741-748
Author(s):  
Mengxi Wang ◽  
Yaoyao Guo ◽  
Yumeng Zhou ◽  
Wanwan Yuan ◽  
Huixia Li ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Biological Activities ◽  
Tumor Formation ◽  
Brown Adipocyte ◽  
Brown Adipocytes ◽  
Protein Levels ◽  
Infected Cells ◽  
Oil Red O Staining ◽  
Oil Red O

AbstractOsteopontin (OPN), a secreted glycoprotein, is involved in various pathophysiological processes including immune response, inflammation, tumor formation, and metabolism. OPN exists in 2 forms, secreted-OPN (sOPN) and intracellular-OPN (iOPN). While they might have different biological activities, it remains largely unknown whether sOPN and iOPN induce the differentiation of brown adipocytes. To test this possibility, 3T3-L1 cells were induced by DMI induction with or without recombinant human OPN (rhOPN, 10, 50, 100, 200 μM), respectively. Meanwhile, another batch of 3T3-L1 cells were infected with Ad-GFP-ap2-OPN and followed by DMI differentiation. Subsequently, the infected cells were treated with either anti-CD44 antibody or immunoglobulin G (Ig G). Accumulation of lipid droplets was visualized by Oil red O staining and protein levels were assayed by western blotting analysis. The results showed that sOPN and not rhOPN, notably increased the accumulation of lipid droplets and the expression of brown adipocyte-related genes. Moreover, neutralization of CD44 partially abrogated the effects induced by sOPN. These data demonstrate that sOPN and not rhOPN has the capacity to induce the differentiation of white preadipocytes into brown adipocytes through a CD44-dependent mechanism. The findings might provide a potential target for sOPN to combat obesity.

scholarly journals Role of Esrrg in the fibrate-mediated regulation of lipid metabolism genes in human ApoA-I transgenic mice

2009 ◽  
Vol 10 (3) ◽  
pp. 165-179 ◽  
Author(s):  
D Sanoudou ◽  
A Duka ◽  
K Drosatos ◽  
K C Hayes ◽  
V I Zannis
Keyword(s):  
Lipid Metabolism ◽  
Transgenic Mice ◽  
Lipid Metabolism Genes

scholarly journals LIPID DROPLET SIGNALING IN METABOLIC HEALTH AND AGING

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. 456-456
Author(s):  
Charles Najt
Keyword(s):  
Fatty Acid ◽  
Energy Metabolism ◽  
Lipid Droplets ◽  
Metabolic Dysfunction ◽  
Critical Pathways ◽  
Cellular Energy ◽  
Cellular Energy Metabolism ◽  
Eicosanoid Production ◽  
Key Players

Abstract Lipid droplets (LDs) are neutral lipid rich organelles involved in lipid storage, fatty acid trafficking, and signaling. Emerging evidence from our laboratory and others suggests that the specific LD resident proteins couple/uncouple cells and tissues from inflammation and metabolic dysfunction. However, the mechanism by which LD proteins influences these critical pathways remains unknown. We will present data delving into the role of LD proteins Perilipin (PLIN) 2 and 5 in balancing cellular energy metabolism, mitochondrial function, and inflammation. Data will be presented defining novel mechanisms through which PLIN2 orchestrates eicosanoid production as a means to promote inflammation. We will contrast these findings to PLIN5, which uncouples LD accumulation from metabolic dysfunction and inflammation, in part due to its promotion of SIRT1 signaling. Overall, these studies will highlight a crucial role of LD metabolism and signaling in regulating cellular energy homeostatic processes known to be key players in governing healthspan.

Sign in / Sign up

Export Citation Format

Share Document