scholarly journals Lipid rafts increase to facilitate ectoderm lineage specification of differentiating embryonic stem cells

2019 ◽  
Author(s):  
Chen Xu ◽  
Bo Cao ◽  
Ying-dong Huo ◽  
Gang Niu ◽  
Michael Q Zhang ◽  
...  
Keyword(s):  
Lipid Rafts ◽  
Early Stage ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Super Resolution ◽  
Lineage Specification ◽  
Genetic Ablation ◽  
Differentiated Cells ◽  
Extracellular Stimuli ◽  
Optical Reconstruction

AbstractLipid rafts are packed nanoscopic domains on plasma membrane and essential signalling platforms for transducing extracellular stimuli into cellular responses. Although depletion of raft component glycoshpingolipids causes abnormality particularly in ectoderm layer formation, it remains unclear whether rafts play a role in lineage determination, a critical but less-known stage in lineage commitment. Here, inducing mouse embryonic stem cell (mESC) differentiation with retinoic acid (RA), we observed lipid rafts increased since early stage, especially in ectoderm-like cells. Stochastic optical reconstruction microscopy characterized at super-resolution the distinct raft features in mESCs and the derived differentiated cells. Furthermore, RA-induced commitment of ectoderm-like cells was significantly diminished not only by genetic ablation of rafts but by applying inhibitor for glycosphingolipids or cholesterol at early differentiation stages. Meanwhile, raft inhibition delayed RA-induced pluripotency exit, an early step required for differentiation. Therefore, lipid rafts increase and facilitate ectoderm lineage specification as well as pluripotency exit during mESC differentiation.

scholarly journals Constitutive Expression of GATA4 Dramatically Increases the Cardiogenic Potential of D3 Mouse Embryonic Stem Cells

2016 ◽  
Vol 10 (1) ◽  
pp. 248-257 ◽  
Author(s):  
Lillian L. Laemmle ◽  
Justus B. Cohen ◽  
Joseph C. Glorioso
Keyword(s):  
Cell Line ◽  
Control Cell ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Constitutive Expression ◽  
Mouse Embryonic Stem Cell ◽  
Embryoid Bodies ◽  
Cardiomyocyte Differentiation ◽  
Differentiated Cells ◽  
Cardiac Gene

The transcription factor GATA binding protein 4 (GATA4) is a vital regulator of cardiac programming that acts by inducing the expression of many different genes involved in cardiomyogenesis. Here we generated a D3 mouse embryonic stem cell line that constitutively expresses high levels of GATA4 and show that these cells have dramatically increased cardiogenic potential compared to an eGFP-expressing control cell line. Embryoid bodies (EB) derived from the D3-GATA4 line displayed increased levels of cardiac gene expression and showed more abundant cardiomyocyte differentiation than control eGFP EB. These cells and two additional lines expressing lower levels of GATA4 provide a platform to screen previously untested cardiac genes and gene combinations for their ability to further increase the efficiency of cardiomyocyte differentiation beyond that achieved by transgenic GATA4 alone. Non-integrative delivery of identified gene combinations will aid in the production of differentiated cells for the treatment of ischemic cardiomyopathy.

Abstract 692: Class II HDACs Are Required During Early Stage Mouse Embryonic Stem Cell Differentiation in Response to Nitric Oxide

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Barbara Illi ◽  
Francesco Spallotta ◽  
Jessica Rosati ◽  
Stefania Mattiussi ◽  
Simona Nanni ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Early Stage ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Stem Cell Differentiation ◽  
Class Ii ◽  
Leukaemia Inhibitory Factor ◽  
Common Mechanism ◽  
Embryonic Stem Cell Differentiation ◽  
No Donor

INTRODUCTION & BACKGROUND: We have recently described that in mouse embryonic stem cells (ES) laminar shear stress provides chromatin programming signals. This effect is similar to that of nitric oxide (NO) which directly promotes cardiomyogenesis underlying the possible presence of a common mechanism of action the characterization of which has been the object of the present study. METHODS & RESULTS: ES were treated with the NO donor DETANO from 1 to 24 hours (h) in the presence of absence of leukaemia inhibitory factor (LIF). In LIF-deprived cells the NO donor determined a marked global reduction in histone H3 acetylation paralleled by the expression of a number of cardiovascular markers. In this condition the stemness-associated gene product Oct4 decreased rapidly at mRNA and protein level underlying the potential opening of a NO-dependent differentiation program. Further class-II histone deacetylase (HDAC) 4, 5, and 7, rapidly shuttled to nucleus upon LIF deprivation. This phenomenon was transient as HDAC molecules returned to the cytoplasm in about 6 h. Intriguingly, the exposure of ES to the NO donor allowed the nuclear retention of these enzymes still detectable at the 6 h time-point. Further, we found that NO treatment stimulated the formation of nuclear complexes including the class II HDACs 4, 5 and the protein phosphatase 2A. Remarkably, ES treatment with the class II selective inhibitor MC 1568 prevented ES differentiation in the presence of NO. CONCLUSIONS: These results suggest a potential role for class II HDACs in the NO-dependent early stage of ES commitment upon LIF removal.

Mechanistic insights into EGFR membrane clustering revealed by super-resolution imaging

Nanoscale ◽  
2015 ◽  
Vol 7 (6) ◽  
pp. 2511-2519 ◽  
Author(s):  
Jing Gao ◽  
Ye Wang ◽  
Mingjun Cai ◽  
Yangang Pan ◽  
Haijiao Xu ◽  
...  
Keyword(s):  
Distribution Pattern ◽  
Lipid Rafts ◽  
Essential Role ◽  
Super Resolution ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Polarized Cells ◽  
Resolution Imaging ◽  
Optical Reconstruction ◽  
Cluster Maintenance

We investigate the distribution of membrane EGFR by direct stochastic optical reconstruction microscopy (dSTORM). Our results illustrate the clustering distribution pattern of EGFR in polarized cells and uncover the essential role of lipid rafts in EGFR cluster maintenance.

scholarly journals H3K9me3-heterochromatin loss at protein-coding genes enables developmental lineage specification

Science ◽  
2019 ◽  
Vol 363 (6424) ◽  
pp. 294-297 ◽  
Author(s):  
Dario Nicetto ◽  
Greg Donahue ◽  
Tanya Jain ◽  
Tao Peng ◽  
Simone Sidoli ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Cell Number ◽  
Specific Gene ◽  
Cell Fates ◽  
Lineage Specification ◽  
Protein Coding ◽  
Protein Coding Genes ◽  
Differentiated Cells ◽  
Cell Type Specific ◽  
Embryologic Development

Gene silencing by chromatin compaction is integral to establishing and maintaining cell fates. Trimethylated histone 3 lysine 9 (H3K9me3)–marked heterochromatin is reduced in embryonic stem cells compared to differentiated cells. However, the establishment and dynamics of closed regions of chromatin at protein-coding genes, in embryologic development, remain elusive. We developed an antibody-independent method to isolate and map compacted heterochromatin from low–cell number samples. We discovered high levels of compacted heterochromatin, H3K9me3-decorated, at protein-coding genes in early, uncommitted cells at the germ-layer stage, undergoing profound rearrangements and reduction upon differentiation, concomitant with cell type–specific gene expression. Perturbation of the three H3K9me3-related methyltransferases revealed a pivotal role for H3K9me3 heterochromatin during lineage commitment at the onset of organogenesis and for lineage fidelity maintenance.

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