scholarly journals Embryonal carcinoma cells transfected with ZP3 genes differentially glycosylate similar polypeptides and secrete active mouse sperm receptor.

1991 ◽  
Vol 115 (3) ◽  
pp. 655-664 ◽  
Author(s):  
R A Kinloch ◽  
S Mortillo ◽  
C L Stewart ◽  
P M Wassarman
Keyword(s):  
Acrosome Reaction ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Transfected Cells ◽  
Mouse Sperm ◽  
Large Numbers ◽  
Ec Cells ◽  
Sperm Receptor

Mouse and hamster sperm receptors, called mZP3 (approximately 83,000 Mr) and hZP3 (approximately 56,000 Mr), respectively, are glycoproteins located in the ovulated egg zona pellucida. Certain of the glycoprotein O-linked oligosaccharides are essential for sperm receptor activity. Here, we transfected mouse embryonal carcinoma (EC) cells with mZP3 and hZP3 genes placed under control of a constitutive promoter. Transfected cells synthesized and secreted large amounts of the glycoproteins, called EC-mZP3 and EC-hZP3. Although the primary structures of mZP3 and hZP3 polypeptides (44,000 Mr) are very similar to one another, EC-mZP3 (approximately 83,000 Mr) and EC-hZP3 (approximately 49,000 Mr) were glycosylated to very different extents, such that they resembled their egg counterparts. Like egg mZP3, EC-mZP3 inhibited binding of sperm to ovulated eggs and induced sperm to acrosome-react in vitro. In addition, large numbers of sperm bound to aggregates of mZP3-transfected EC cells in vitro. On the other hand, unlike egg hZP3, EC-hZP3 did not exhibit either sperm receptor or acrosome reaction-inducing activity, and sperm failed to bind to aggregates of hZP3-transfected EC cells. Thus, transfected EC cells not only express sperm receptor genes, but also discriminate between very similar polypeptides with respect to glycosylation and, in the case of mZP3, add specific oligosaccharides essential for biological activity. In addition, the results demonstrate that EC cells can serve as a source for large amounts of functional mouse sperm receptor.

scholarly journals SNF2beta-BRG1 is essential for the viability of F9 murine embryonal carcinoma cells.

1997 ◽  
Vol 17 (10) ◽  
pp. 5976-5986 ◽  
Author(s):  
C Sumi-Ichinose ◽  
H Ichinose ◽  
D Metzger ◽  
P Chambon
Keyword(s):  
Transcriptional Activation ◽  
Embryonal Carcinoma ◽  
Regulatory Elements ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Nucleosome Remodeling ◽  
Transfected Cells ◽  
Activation Of Transcription ◽  
Ec Cells ◽  
Mutant Cells

The yeast and animal SNF-SWI and related multiprotein complexes are thought to play an important role in processes, such as transcription factor binding to regulatory elements, which require nucleosome remodeling in order to relieve the repressing effect of packaging DNA in chromatin. There are two mammalian homologs of the yeast SNF2-SWI2 subunit protein, SNF2alpha-brm and SNF2beta-BRG1, and overexpression of either one of them has been shown to enhance transcriptional activation by glucocorticoid, estrogen, and retinoic acid (RA) receptors in transiently transfected cells. We have investigated here the function of SNF2beta-BRG1 in the RA receptor-retinoid X receptor-mediated transduction of the retinoid signal in F9 embryonal carcinoma (EC) cells which differentiate into endodermal-like cells upon RA treatment. The two SNF2beta-BRG1 alleles have been targeted by homologous recombination and subsequently disrupted by using a conditional Cre recombinase. We show that F9 EC cells inactivated on both SNF2beta alleles are not viable and that heterozygous mutant cells are affected in proliferation but not in RA-induced differentiation. Thus, in F9 EC cells, SNF2beta-BRG1 appears to play an essential role in basal processes involved in cell proliferation, in addition to its putative role in the activation of transcription mediated by nuclear receptors.

scholarly journals Apoptosis of Human Embryonal Carcinoma Cells with in vitro Differentiation.

1996 ◽  
Vol 21 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Taketo Yamada ◽  
Nao Suzuki ◽  
Nobuyoshi Hiraoka ◽  
Kentaroh Matsuoka ◽  
Sachiko Fukushima ◽  
...  
Keyword(s):  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
In Vitro Differentiation

scholarly journals Demethylation of somatic and testis-specific histone H2A and H2B genes in F9 embryonal carcinoma cells

1993 ◽  
Vol 13 (9) ◽  
pp. 5538-5548
Author(s):  
Y C Choi ◽  
C B Chae
Keyword(s):  
Embryonal Carcinoma ◽  
Cpg Island ◽  
Cpg Islands ◽  
High Density ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Cpg Dinucleotides ◽  
F9 Embryonal Carcinoma Cells ◽  
Methylation Patterns

In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became significantly inefficient at a high number of integrated copies and a high density of methylated CpG dinucleotides. In contrast, three sites in the somatic histone cluster, of which two sites are located in the long terminal repeat of an endogenous retrovirus-like sequence, were efficiently demethylated even at a high copy number and a high density of methylated CpG dinucleotides. These results suggest two possible mechanisms for demethylation in F9 cells and methylation of CpG islands of the TH2A and TH2B genes at the postblastula stage during embryogenesis.

scholarly journals Demethylation of somatic and testis-specific histone H2A and H2B genes in F9 embryonal carcinoma cells.

1993 ◽  
Vol 13 (9) ◽  
pp. 5538-5548 ◽  
Author(s):  
Y C Choi ◽  
C B Chae
Keyword(s):  
Embryonal Carcinoma ◽  
Cpg Island ◽  
Cpg Islands ◽  
High Density ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Cpg Dinucleotides ◽  
F9 Embryonal Carcinoma Cells ◽  
Methylation Patterns

In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became significantly inefficient at a high number of integrated copies and a high density of methylated CpG dinucleotides. In contrast, three sites in the somatic histone cluster, of which two sites are located in the long terminal repeat of an endogenous retrovirus-like sequence, were efficiently demethylated even at a high copy number and a high density of methylated CpG dinucleotides. These results suggest two possible mechanisms for demethylation in F9 cells and methylation of CpG islands of the TH2A and TH2B genes at the postblastula stage during embryogenesis.

scholarly journals Ectopic activation of WNT signaling in human embryonal carcinoma cells and its effects in short- and long-term in vitro culture

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yaser Atlasi ◽  
Rebecca T. van Dorsten ◽  
Andrea Sacchetti ◽  
Rosalie Joosten ◽  
J. Wolter Oosterhuis ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
In Vitro Culture ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Ectopic Activation ◽  
Short And Long Term

Sperm Performance Better on Diamond than on Polystyrene

2013 ◽  
Vol 1511 ◽  
Author(s):  
Andrei P. Sommer ◽  
Dan Zhu ◽  
Friedrich Gagsteiger ◽  
Hans-Jörg Fecht
Keyword(s):  
Embryonal Carcinoma ◽  
Petri Dish ◽  
Carcinoma Cells ◽  
Oxygen Species ◽  
Embryonal Carcinoma Cells ◽  
Vitro Fertilization ◽  
Polystyrene Petri Dish ◽  
Petri Dishes ◽  
New Generation

ABSTRACTRecently we postulated that polystyrene Petri dishes become soft when in contact with an aqueous milieu. Specifically, we assumed that the effect is restricted to a superficial nanolayer, a condition presumably favoring the establishment of a stable nanolayer of reactive oxygen species (ROS) at the liquid/solid-interface. Cells are known to be hypersensitive to ROS. Previously we used P19 mouse embryonal carcinoma cells and systematically analyzed their capability to climb different substrates placed vertically into a Petri dish. The worst and best performance was found on polystyrene (Petri dish material) and nanocrystalline diamond, respectively. Polystyrene Petri dishes are today standard in laboratories conducting in vitro fertilization (IVF). Here we proceed and extend the investigation to human spermatozoa and show that their performance (vitality) on polystyrene Petri dishes is low compared to that on diamond Petri dishes. This work may propel further research and inspire the development of a new generation of cell-friendly Petri dishes.

scholarly journals Multiple CArG boxes in the human cardiac actin gene promoter required for expression in embryonic cardiac muscle cells developing in vitro from embryonal carcinoma cells.

1991 ◽  
Vol 11 (9) ◽  
pp. 4796-4803 ◽  
Author(s):  
G Pari ◽  
K Jardine ◽  
M W McBurney
Keyword(s):  
Cardiac Muscle ◽  
Embryonal Carcinoma ◽  
Core Promoter ◽  
Carcinoma Cells ◽  
Regulatory Sequences ◽  
Embryonal Carcinoma Cells ◽  
Cardiac Muscle Cells ◽  
Cardiac Actin ◽  
Actin Promoter

Chimeric genes composed of the human cardiac actin promoter driving the Escherichia coli lacZ reporter gene were constructed, transfected, and stably integrated into genomes of P19 embryonal carcinoma cells. The transfected constructs were expressed actively in cardiac myocytes formed following dimethyl sulfoxide (DMSO)-induced cell differentiation but poorly in undifferentiated cultures and in cultures treated with retinoic acid to develop into derivatives of the neuroectoderm. A number of deletions of the promoter were constructed and tested. Three regions required for efficient expression in P19-derived cardiac muscle were identified, each containing sequences referred to as CArG boxes (CC[AT-rich]6GG). This analysis indicated that regulatory sequences important for expression in cardiac muscle were present upstream of the core promoter identified previously by transient assays in skeletal myoblasts. Expression of the cardiac actin promoter was enhanced 10-fold in undifferentiated P19 cells in the presence of the myoD protein. The promoter regions important for expression in P19-derived cardiocytes were similar to those important for myoD-induced enhancement, a result we interpret to be consistent with the idea that cardiac muscle might contain a myoD-like activity.

Mechanism of acetylcholine-induced calcium signaling during neuronal differentiation of P19 embryonal carcinoma cells in vitro

Cell Calcium ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 107-121 ◽  
Author(s):  
Rodrigo R. Resende ◽  
Katia N. Gomes ◽  
Avishek Adhikari ◽  
Luiz R.G. Britto ◽  
Henning Ulrich
Keyword(s):  
Calcium Signaling ◽  
Neuronal Differentiation ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
P19 Embryonal Carcinoma Cells

Specific high-affinity binding of 125I-labeled mouse interferon to interferon resistant embryonal carcinoma cells in vitro

Virology ◽  
1981 ◽  
Vol 114 (2) ◽  
pp. 585-588 ◽  
Author(s):  
Michel Aguet ◽  
Ion Gresser ◽  
Ara G. Hovanessian ◽  
Marie-Thérèse Bandu ◽  
Brigitte Blanchard ◽  
...  
Keyword(s):  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Affinity Binding ◽  
Embryonal Carcinoma Cells ◽  
High Affinity Binding ◽  
High Affinity
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