FTO and ALKBH5 proteins are essential erasers of N6-adenosine methylation in RNA. We studied how levels of FTO and ALKBH5 proteins changed during mouse embryonic development, aging, cardiomyogenesis, and neuroectodermal differentiation. We observed that aging in male and female mice was associated with FTO up-regulation in mouse hearts, brains, lungs, and kidneys, while the ALKBH5 level remained stable. FTO and ALKBH5 proteins were up-regulated during experimentally induced cardiomyogenesis, but the level of ALKBH5 protein was not changed when neuroectodermal differentiation was induced. HDAC1 depletion in mouse ES cells caused FTO down-regulation. In these cells, mRNA, carrying information from genes that regulate histone signature, RNA processing, and cell differentiation, was characterized by a reduced level of N6-adenosine methylation in specific gene loci, primarily regulating cell differentiation into neuroectoderm. Together, when we compared both RNA demethylating proteins, the FTO protein level undergoes the most significant changes during cell differentiation and aging. Thus, we conclude that during aging and neuronal differentiation, m6A RNA demethylation is likely regulated by the FTO protein but not via the function of ALKBH5.

• MeSH
• Adenosine metabolism   MeSH
• AlkB Homolog 5, RNA Demethylase * genetics   metabolism   MeSH
• Cell Differentiation MeSH
• Embryonic Development MeSH
• Alpha-Ketoglutarate-Dependent Dioxygenase FTO * genetics   metabolism   MeSH
• Mice MeSH
• RNA metabolism   MeSH
• Aging genetics   MeSH
• Up-Regulation MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Adenosine MeSH
• AlkB Homolog 5, RNA Demethylase * MeSH
• FTO protein, mouse MeSH Browser
• Alpha-Ketoglutarate-Dependent Dioxygenase FTO * MeSH
• RNA MeSH

The active role of biomaterials in the regeneration of tissues and their ability to modulate the behavior of stem cells in terms of their differentiation is highly advantageous. Here, polypyrrole, as a representantive of electro-conducting materials, is found to modulate the behavior of embryonic stem cells. Concretely, the aqueous extracts of polypyrrole induce neurogenesis within embryonic bodies formed from embryonic stem cells. This finding ledto an effort to determine the physiological cascade which is responsible for this effect. The polypyrrole modulates signaling pathways of Akt and ERK kinase through their phosphorylation. These effects are related to the presence of low-molecular-weight compounds present in aqueous polypyrrole extracts, determined by mass spectroscopy. The results show that consequences related to the modulation of stem cell differentiation must also be taken into account when polypyrrole is considered as a biomaterial.

• Keywords
• biocompatibility, conducting polymer, neurogenesis, polypyrrole, stem cells,
• MeSH
• Cell Differentiation drug effects   genetics   MeSH
• Cell Line MeSH
• Embryoid Bodies cytology   drug effects   MeSH
• Gene Expression drug effects   MeSH
• Molecular Structure MeSH
• Mouse Embryonic Stem Cells cytology   drug effects   metabolism   MeSH
• Mice MeSH
• Neural Stem Cells cytology   drug effects   metabolism   MeSH
• Neurogenesis drug effects   genetics   MeSH
• Reverse Transcriptase Polymerase Chain Reaction MeSH
• Polymers chemistry   pharmacology   MeSH
• Pyrroles chemistry   pharmacology   MeSH
• PAX6 Transcription Factor genetics   MeSH
• Basic Helix-Loop-Helix Transcription Factors genetics   MeSH
• SOXB1 Transcription Factors genetics   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Ascl1 protein, mouse MeSH Browser
• Polymers MeSH
• polypyrrole MeSH Browser
• Pyrroles MeSH
• PAX6 Transcription Factor MeSH
• Basic Helix-Loop-Helix Transcription Factors MeSH
• SOXB1 Transcription Factors MeSH

Although histone acetylation is one of the most widely studied epigenetic modifications, there is still a lack of information regarding how the acetylome is regulated during brain development and pathophysiological processes. We demonstrate that the embryonic brain (E15) is characterized by an increase in H3K9 acetylation as well as decreases in the levels of HDAC1 and HDAC3. Moreover, experimental induction of H3K9 hyperacetylation led to the overexpression of NCAM in the embryonic cortex and depletion of Sox2 in the subventricular ependyma, which mimicked the differentiation processes. Inducing differentiation in HDAC1-deficient mouse ESCs resulted in early H3K9 deacetylation, Sox2 downregulation, and enhanced astrogliogenesis, whereas neuro-differentiation was almost suppressed. Neuro-differentiation of (wt) ESCs was characterized by H3K9 hyperacetylation that was associated with HDAC1 and HDAC3 depletion. Conversely, the hippocampi of schizophrenia-like animals showed H3K9 deacetylation that was regulated by an increase in both HDAC1 and HDAC3. The hippocampi of schizophrenia-like brains that were treated with the cannabinoid receptor-1 inverse antagonist AM251 expressed H3K9ac at the level observed in normal brains. Together, the results indicate that co-regulation of H3K9ac by HDAC1 and HDAC3 is important to both embryonic brain development and neuro-differentiation as well as the pathophysiology of a schizophrenia-like phenotype.

• Keywords
• H3K9 acetylation, HDACs, acetylome, mouse neurogenesis, schizophrenia,
• MeSH
• Acetylation MeSH
• Cannabinoid Receptor Antagonists pharmacology   MeSH
• Antipsychotic Agents pharmacology   MeSH
• Time Factors MeSH
• Epigenesis, Genetic MeSH
• Gestational Age MeSH
• Histone Deacetylase 1 antagonists & inhibitors   genetics   metabolism   MeSH
• Histone Deacetylases genetics   metabolism   MeSH
• Histones metabolism   MeSH
• Histone Deacetylase Inhibitors pharmacology   MeSH
• Methylazoxymethanol Acetate MeSH
• Disease Models, Animal MeSH
• Neural Cell Adhesion Molecules genetics   metabolism   MeSH
• Brain drug effects   embryology   enzymology   pathology   MeSH
• Mice, Inbred C57BL MeSH
• Neurogenesis * drug effects   MeSH
• Neurons drug effects   enzymology   pathology   MeSH
• Protein Processing, Post-Translational MeSH
• Rats, Sprague-Dawley MeSH
• Receptor, Cannabinoid, CB1 antagonists & inhibitors   metabolism   MeSH
• Schizophrenia chemically induced   drug therapy   enzymology   genetics   MeSH
• Signal Transduction MeSH
• SOXB1 Transcription Factors genetics   metabolism   MeSH
• Gene Expression Regulation, Developmental MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Cannabinoid Receptor Antagonists MeSH
• Antipsychotic Agents MeSH
• Cnr1 protein, rat MeSH Browser
• Hdac1 protein, mouse MeSH Browser
• Hdac1 protein, rat MeSH Browser
• Histone Deacetylase 1 MeSH
• Histone Deacetylases MeSH
• histone deacetylase 3 MeSH Browser
• Histones MeSH
• Histone Deacetylase Inhibitors MeSH
• Methylazoxymethanol Acetate MeSH
• Neural Cell Adhesion Molecules MeSH
• Receptor, Cannabinoid, CB1 MeSH
• Sox2 protein, mouse MeSH Browser
• SOXB1 Transcription Factors MeSH

The differentiation of pluripotent embryonic stem (ES) cells into various lineages in vitro represents an important tool for studying the mechanisms underlying mammalian embryogenesis. It is a key technique in studies evaluating the molecular mechanisms of cardiomyogenesis and heart development and also in embryotoxicology. Herein, modest modifications of the basic protocol for ES cell differentiation into cardiomyocytes were evaluated in order to increase the yield and differentiation status of developed cardiomyocytes. Primarily, the data show that ES cell cultivation in the form of non-adherent embryoid bodies (EBs) for 5 days compared to 8 days significantly improved cardiomyogenic differentiation. This is illustrated by the appearance of beating foci in the adherent EBs layer at earlier phases of differentiation from day 10 up to day 16 and by the significantly higher expression of genes characteristic of cardiomyogenic differentiation (sarcomeric alpha actinin, myosin heavy chain alpha and beta, myosin light chain 2 and 7, and transcriptional factor Nkx2.5) in EBs cultivated under non-adherent conditions for 5 days. The ratio of cardiomyocytes per other cells was also potentiated in EBs cultivated in non-adherent conditions for only 5 days followed by cultivation in adherent serum-free culture conditions. Nevertheless, the alteration in the percentage of beating foci among these two tested cultivation conditions vanished at later phases and also did not affect the total number of cardiomyocytes determined as myosin heavy chain positive cells at the end of the differentiation process on day 20. Thus, although these modifications of the conditions of ES cells differentiation may intensify cardiomyocyte differentiation, the final count of cardiomyocytes might not change. Thus, serum depletion was identified as a key factor that intensified cardiomyogenesis. Further, the treatment of EBs with N-acetylcysteine, a reactive oxygen species scavenger, did not affect the observed increase in cardiomyogenesis under serum depleted conditions. Interestingly, a mild induction of the ventricular-like phenotype of cardiomyocytes was observed in 5-day-old EBs compared to 8-day-old EBs. Overall, these findings bring crucial information on the mechanisms of ES cells differentiation into cardiomyocytes and on the establishment of efficient protocols for the cardiomyogenic differentiation of ES cells. Further, the importance of determining the absolute number of formed cardiomyocyte-like cells per seeded pluripotent cells in contrast to the simple quantification of the ratios of cells is highlighted.

• MeSH
• Acetylcysteine administration & dosage   MeSH
• Actinin genetics   MeSH
• Embryonic Stem Cells cytology   MeSH
• Homeobox Protein Nkx-2.5 genetics   MeSH
• Myocytes, Cardiac cytology   MeSH
• Culture Media, Serum-Free * MeSH
• Cells, Cultured MeSH
• Myosins genetics   MeSH
• Mice MeSH
• In Vitro Techniques MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Acetylcysteine MeSH
• Actinin MeSH
• Homeobox Protein Nkx-2.5 MeSH
• Culture Media, Serum-Free * MeSH
• Myosins MeSH
• Nkx2-5 protein, mouse MeSH Browser

Cajal bodies (CBs) are important compartments containing accumulated proteins that preferentially regulate RNA-related nuclear events, including splicing. Here, we studied the nuclear distribution pattern of CBs in neurogenesis. In adult brains, coilin was present at a high density, but CB formation was absent in the nuclei of the choroid plexus of the lateral ventricles. Cells of the adult hippocampus were characterized by a crescent-like morphology of coilin protein. We additionally observed a 70 kDa splice variant of coilin in adult mouse brains, which was different to embryonic brains and mouse pluripotent embryonic stem cells (mESCs), characterized by the 80 kDa standard variant of coilin. Here, we also showed that depletion of coilin is induced during neural differentiation and HDAC1 deficiency in mESCs caused coilin accumulation inside the fibrillarin-positive region of the nucleoli. A similar distribution pattern was observed in adult brain hippocampi, characterized by lower levels of both coilin and HDAC1. In summary, we observed that neural differentiation and HDAC1 deficiency lead to coilin depletion and coilin accumulation in body-like structures inside the nucleoli.

• Publication type
• Journal Article MeSH

Alkaline phosphatase is an enzyme commonly expressed in almost all living organisms. In humans and other mammals, determinations of the expression and activity of alkaline phosphatase have frequently been used for cell determination in developmental studies and/or within clinical trials. Alkaline phosphatase also seems to be one of the key markers in the identification of pluripotent embryonic stem as well as related cells. However, alkaline phosphatases exist in some isoenzymes and isoforms, which have tissue specific expressions and functions. Here, the role of alkaline phosphatase as a stem cell marker is discussed in detail. First, we briefly summarize contemporary knowledge of mammalian alkaline phosphatases in general. Second, we focus on the known facts of its role in and potential significance for the identification of stem cells.

• Publication type
• Journal Article MeSH
• Review MeSH