Diverse effects of essential (n?6 andn?3) fatty acids on cultured cells

1994 ◽  
Vol 15 (1-3) ◽  
pp. 31-50 ◽  
Author(s):  
Stephanos I. Grammatikos ◽  
Papasani V. Subbaiah ◽  
Thomas A. Victor ◽  
William M. Miller
Keyword(s):  
Fatty Acids ◽  
Cultured Cells

scholarly journals Phytohormones Regulate the Non-Redundant Response of ω-3 Fatty Acid Desaturases to Low Temperatures in Chorispora Bungeana

2021 ◽  
Author(s):  
Yulan Shi ◽  
Sizhong Yang ◽  
Xiule Yue ◽  
Zhixing Zhao ◽  
Lizhe An
Keyword(s):  
Fatty Acids ◽  
Abscisic Acid ◽  
Fatty Acid ◽  
Stress Response ◽  
Low Temperatures ◽  
Cultured Cells ◽  
Temperature Decrease ◽  
Fatty Acid Desaturases ◽  
Cold Environments ◽  
Chorispora Bungeana

Abstract To explore the contribution of ω-3 fatty acid desaturases (FADs) to cold stress response in a special cryophyte, Chorispora bungeana (C. bungeana), two plastidial ω-3 FAD genes (CbFAD7 and CbFAD8) were cloned and verified in a Arabidopsis fad7fad8 mutant, before being compared with the microsomal ω-3 FAD gene (CbFAD3) on expression profile. Though these genes were expressed in all tested tissues of C. bungeana, CbFAD7 and CbFAD8 have the highest expression in leaves, while CbFAD3 was mostly expressed in non-green tissues. Low temperatures (4, 0 and -4 ℃) resulted in significant increases in trienoic fatty acids (TAs, mainly C18:3), which were consistent with the non-redundant expression of CbFAD3 and CbFAD8 in suspension-cultured cells, and the coordination of CbFAD7 and CbFAD8 in leaves. Furthermore, the contribution of CbFAD8 increased as temperature decrease in the two tissues. Our data revealed that jasmonie acid and brassinosteroids participated in the cold-responsive expression of these genes in both tissues, and the pyhtohormone regulation in leaves was more complicated with the participation of abscisic acid and gibberellin. These results point to the hormone-regulated non-redundant contribution of ω-3 CbFADs to maintain appropriate level of TAs under low temperatures, which help C. bungeana survive in cold environments.

Diverse effects of essential (n-6 and n-3) fatty acids on cultured cells

1994 ◽  
pp. 31-50 ◽  
Author(s):  
Stephanos I. Grammatikos ◽  
Papasani V. Subbaiah ◽  
Thomas A. Victor ◽  
William M. Miller
Keyword(s):  
Fatty Acids ◽  
Cultured Cells

34 PRODUCTION OF TRANSGENIC CLONED BUFFALO EMBRYOS CONTAINING OVEREXPRESSED STEAROYL Co-A DESATURASE GENE FOLLOWING EFFICIENT TRANSFECTION

2017 ◽  
Vol 29 (1) ◽  
pp. 124
Author(s):  
T. Sharma ◽  
D. Dua ◽  
N. Saini ◽  
M. K. Singh ◽  
S. K. Singla ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Nuclear Transfer ◽  
Cultured Cells ◽  
Saturated Fatty Acids ◽  
Phospholipid Composition ◽  
Calcium Ionophore ◽  
Basal Medium ◽  
Developmental Competence ◽  
Fetal Fibroblast ◽  
Gene Level

Stearoyl-CoA desaturase (SCD) is a rate-limiting enzyme that catalyses the synthesis of monounsaturated fatty acids and polyunsaturated fatty acids from saturated fatty acids, which are components of triglycerides, wax esters, cholesteryl esters, and membrane phospholipids. Alterations in phospholipid composition have been implicated in a variety of diseases including obesity and the associated metabolic syndrome. SCD also magnifies the conjugated linoleic acid (CLA) content in milk; CLA is a natural fat element, having reputed therapeutic health values including anti-carcinogenic properties. In light of this fact, this study was designed to amplify the levels of SCD at the gene level. In order to achieve the enhanced expression of SCD gene, combination of techniques were used. The somatic cells (fetal fibroblast) culture were established from ear pinnae obtained from bovine fetus procured from the abattoir and were cultured in basal medium, comprising DMEM with 10% fetal bovine serum, 1X NEAA and 1X PS antibiotics. These isolated cultured cells were transfected with a gene construct carrying buSCD gene (pAcGFPN1-buSCD) as BLGP-buSCD-BLG3′UTR-CMVP-EGFP-SV40. The buffalo fetal fibroblast cells were transfected using 3 methods: Nucleofection, Fugene and Lipofection. The successful transfection, as confirmed by PCR and Southern hybridisation, proved Nucleofection to be more efficient in transfecting cells among the techniques used, which were further maintained and selected by Geneticin (G418). These selected transfected cells were then used for nuclear transfer. Somatic cell nuclear transfer (SCNT) has provided an efficient pathway for the production of transgenic animals. Buffalo cumulus–oocyte complexes (COCs) were collected from ovaries collected from abattoir and matured in TCM-199 supplemented with 10% FBS, 5 µg mL−1 FSH, and 1 µg mL−1 β-oestradiol for 21 h in CO2 incubator at 5% CO2 in air and 38.5°C temperature with >95% relative humidity. After 21 h, these COCs were denuded and subjected to zona removal. These zona-free oocytes were manually enucleated using microsurgical blades and 2 enucleated oocytes were fused with a transgenic cell using electro cell manipulator. Further, these reconstructed embryos were activated using calcium ionophore and cultured in IVC media thereafter for 8 days. The developmental competence rate as recorded on Day 8 was 53.26 ± 1.73%, 69.87 ± 6.24%, 62.99 ± 7.15%, 42.71 ± 5.05% and 28.00 ± 3.33% for 2-cell, 4-cell, 8–16 cell, morula, and blastocyst, respectively. When observed under fluorescence microscope, the embryos showed successful expression of GFP, which can be further used for animal production or further research analysis. In conclusion, amplified SCD at gene level will result in a boost to the dairy sector as well ameliorating human health due to its crucial role in anti-cancer, anti-diabetic, reduced cardio-vascular disease, and improved immune responses.

scholarly journals Unsaturated Fatty Acids Phosphorylate and Destabilize ABCA1 through a Phospholipase D2 Pathway

2005 ◽  
Vol 280 (43) ◽  
pp. 35896-35903 ◽  
Author(s):  
Yutong Wang ◽  
John F. Oram
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Unsaturated Fatty Acids ◽  
Cultured Cells ◽  
Saturated Fatty Acids ◽  
Density Lipoprotein ◽  
Serine Phosphorylation ◽  
Protein Levels ◽  
Increased Risk ◽  
Abca1 Protein

Abnormal high density lipoprotein (HDL) metabolism among patients with diabetes and insulin resistance may contribute to their increased risk of atherosclerosis. ATP-binding cassette transporter ABCA1 mediates the transport of cholesterol and phospholipids from cells to HDL apolipoproteins and thus modulates HDL levels and atherogenesis. Unsaturated fatty acids, which are elevated in diabetes, impair the ABCA1 pathway in cultured cells by destabilizing ABCA1 protein. Here we examined the cellular pathway that mediates the ABCA1 destabilizing effects of fatty acids. The long-chain acyl-CoA synthetase inhibitor triacsin C completely reversed fatty acid-induced ABCA1 destabilization, indicating that fatty acids need to be activated to their CoA derivatives to enhance ABCA1 degradation. Unsaturated but not saturated fatty acids stimulated phospholipase D (PLD) activity, the PLD inhibitor 1-butanol prevented the unsaturated fatty acid-induced reduction in ABCA1 levels, and the PLD2 activator mastoparan markedly reduced ABCA1 protein levels, implicating a role for PLD2 in the ABCA1 destabilizing effects of fatty acids. Unsaturated fatty acids and mastoparan increased phosphorylation of ABCA1 serines. PLD2 small interfering RNA abolished the ability of unsaturated fatty acids to inhibit lipid transport activity, to reduce protein levels, and to increase serine phosphorylation of ABCA1. The diacylglycerol analog oleoylacetylglycerol also reduced ABCA1 protein levels and increased its serine phosphorylation, suggesting that PLD2-generated diacylglycerols promote the destabilizing phosphorylation of ABCA1. These data provide evidence that intracellular unsaturated acyl-CoA derivatives destabilize ABCA1 by activating a PLD2 signaling pathway.

scholarly journals Distinct roles of specific fatty acids in cellular processes: implications for interpreting and reporting experiments

2012 ◽  
Vol 302 (1) ◽  
pp. E1-E3 ◽  
Author(s):  
Matthew J. Watt ◽  
Andrew J. Hoy ◽  
Deborah M. Muoio ◽  
Rosalind A. Coleman
Keyword(s):  
Fatty Acids ◽  
High Fat Diet ◽  
Cultured Cells ◽  
Cellular Function ◽  
Long Chain Fatty Acids ◽  
High Fat ◽  
Cellular Processes ◽  
Stress Effects ◽  
Long Chain ◽  
Chain Fatty Acids

Plasma contains a variety of long-chain fatty acids (FAs), such that about 35% are saturated and 65% are unsaturated. There are countless examples that show how different FAs impart specific and unique effects, or even opposing actions, on cellular function. Despite these differing effects, palmitate (C16:0) is regularly used to represent “FAs” in cell based experiments. Although palmitate can be useful to induce and study stress effects in cultured cells, these effects in isolation are not physiologically relevant to dietary manipulations, obesity, or the consequences of physiological concentrations of FAs. Hence, authors should avoid conclusions that generalize about “FAs” or “saturated FAs” or “high-fat diet” effects if only a single FA was used in the reported experiments.

scholarly journals Serum lipoprotein–derived fatty acids regulate hypoxia-inducible factor

2020 ◽  
Vol 295 (52) ◽  
pp. 18284-18300
Author(s):  
Wei Shao ◽  
Jiwon Hwang ◽  
Chune Liu ◽  
Debaditya Mukhopadhyay ◽  
Shan Zhao ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Transcription Factors ◽  
Target Genes ◽  
Cultured Cells ◽  
Cell Metabolism ◽  
Control Cell ◽  
Hypoxia Inducible Factor ◽  
Mitochondrial Respiratory Chain ◽  
Serum Lipoprotein ◽  
Downstream Target

Oxygen regulates hypoxia-inducible factor (HIF) transcription factors to control cell metabolism, erythrogenesis, and angiogenesis. Whereas much has been elucidated about how oxygen regulates HIF, whether lipids affect HIF activity is un-known. Here, using cultured cells and two animal models, we demonstrate that lipoprotein-derived fatty acids are an independent regulator of HIF. Decreasing extracellular lipid supply inhibited HIF prolyl hydroxylation, leading to accumulation of the HIFα subunit of these heterodimeric transcription factors comparable with hypoxia with activation of downstream target genes. The addition of fatty acids to culture medium suppressed this signal, which required an intact mitochondrial respiratory chain. Mechanistically, fatty acids and oxygen are distinct signals integrated to control HIF activity. Finally, we observed lipid signaling to HIF and changes in target gene expression in developing zebrafish and adult mice, and this pathway operates in cancer cells from a range of tissues. This study identifies fatty acids as a physiological modulator of HIF, defining a mechanism for lipoprotein regulation that functions in parallel to oxygen.

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