A5103071327- Pluripotent Stem Cells Research
- Genomics and Chromatin Dynamics
- CRISPR and Genetic Engineering
- Reproductive Biology and Fertility
- Epigenetics and DNA Methylation
- RNA Research and Splicing
- Animal Genetics and Reproduction
- DNA Repair Mechanisms
- Renal and related cancers
- Prenatal Screening and Diagnostics
- Cancer-related molecular mechanisms research
- Mitochondrial Function and Pathology
- Biochemical Analysis and Sensing Techniques
- Skin Protection and Aging
- melanin and skin pigmentation
- Muscle Physiology and Disorders
- FOXO transcription factor regulation
- Lysosomal Storage Disorders Research
- Genetics and Neurodevelopmental Disorders
- Corneal Surgery and Treatments
- Cancer, Hypoxia, and Metabolism
- Retinal Development and Disorders
- Cervical Cancer and HPV Research
- ATP Synthase and ATPases Research
- Genetic Neurodegenerative Diseases
Yokohama City University
2023-2024
Keio University
2015-2023
Keio University Hospital
2017
National Institute on Aging
2015
The University of Tokyo
2004-2014
National Institutes of Health
2013
Ube Frontier University
2004
Errors in meiotic chromosome segregation are the leading cause of spontaneous abortions and birth defects. Almost all such aneuploidy derives from errors females, with increasing maternal age representing a major risk factor. It was recently reported that histones globally deacetylated mammalian oocytes during meiosis but not mitosis. In present study, inhibition histone deacetylation found to induce fertilized mouse oocytes, which resulted embryonic death utero at an early stage...
Upon fertilization, reprogramming of gene expression is required for embryo development. This step marked by DNA demethylation and changes in histone variant composition. However, little known about the molecular mechanisms causing these their impact on modifications. We examined global deposition replication-dependent H3.1 H3.2 variants replication-independent H3.3 after fertilization mice. showed that H3.3, a euchromatic marker activity, transiently disappears from maternal genome,...
Mouse embryonic stem cells (mESCs) have a remarkable capacity to maintain normal genome stability and karyotype in culture. We previously showed that infrequent bursts of Zscan4 expression (Z4 events) are important for the maintenance telomere length mESCs. However, molecular details Z4 events remain unclear. Here we show involve unexpected transcriptional derepression heterochromatin regions usually silent. During event, see rapid rerepression leading burst transcription coincides with...
The gene expression pattern of differentiated oocytes is reprogrammed into that totipotent preimplantation embryos before and/or after fertilization. To elucidate the mechanisms genome reprogramming, we investigated histone H3 lysine 79 dimethylation (H3K79me2) and trimethylation (H3K79me3) in via immunocytochemistry. In somatic cells oocytes, H3K79me2 was observed throughout genome, whereas H3K79me3 localized pericentromeric heterochromatin regions which there are no active genes. Because...
Histone H2A has several variants, and changes in chromatin composition associated with their replacement might involve structure remodeling. We examined the dynamics of canonical histone its three H2A.X, H2A.Z macroH2A, mouse during oogenesis pre-implantation development when genome remodeling occurs. Immunocytochemistry specific antibodies revealed that, although all variants were deposited nuclei full-grown oocytes, only H2A.X was abundant pronuclei one-cell embryos after fertilization,...
Efficient differentiation of human pluripotent stem cells (hPSCs) into neurons is paramount for disease modeling, drug screening, and cell transplantation therapy in regenerative medicine. In this manuscript, we report the capability five transcription factors (TFs) toward aim: NEUROG1, NEUROG2, NEUROG3, NEUROD1, NEUROD2. contrast to previous methods that have shortcomings their speed efficiency, a cocktail these TFs as synthetic mRNAs can differentiate hPSCs 7 days, judged by calcium...
The role of lipid metabolism in human pluripotent stem cells (hPSCs) is poorly understood. We have used large-scale targeted proteomics to demonstrate that undifferentiated hPSCs express different fatty acid (FA) biosynthesis-related enzymes, including ATP citrate lyase and FA synthase (FASN), than those expressed hPSC-derived cardiomyocytes (hPSC-CMs). Detailed profiling revealed inhibition FASN resulted significant reduction sphingolipids phosphatidylcholine (PC); moreover, we found PC was...
Abstract The development of multicellular organisms is accompanied by reprogramming the epigenome in specific cells, with most cell types becoming fixed after differentiation. Genome-wide DNA methylation occurs primordial germ cells and fertilized eggs during mammalian embryogenesis. 5-methylcytosine (5mC) content thus undergoes a marked decrease paternal pronucleus zygotes. This loss has been thought to be mediated an active demethylation mechanism independent replication required for...
Abstract The derivation of kidney tissues from human pluripotent stem cells (hPSCs) and its application for replacement therapy in end-stage renal disease have been widely discussed. Here we report that consecutive transfections two sets synthetic mRNAs encoding transcription factors can induce rapid efficient differentiation hPSCs into tissues, termed induced nephron-like organoids (iNephLOs). first set - FIGLA, PITX2, ASCL1 TFAP2C, differentiated SIX2 + SALL1 nephron progenitor with 92%...
Histone acetylation is an important epigenetic modification implicated in the regulation of chromatin structure and, subsequently, gene expression. Global histone deacetylation was reported mouse oocytes during meiosis but not mitosis. The this meiosis-specific has been elucidated. Here, we demonstrate that p34(cdc2) kinase activity and protein synthesis are responsible for activation deacetylases inhibition acetyltransferases (HATs), respectively, resulting H4 at lysine-12 (H4K12) oocyte...
Harnessing epigenetic regulation is crucial for the efficient and proper differentiation of pluripotent stem cells (PSCs) into desired cell types. Histone H3 lysine 27 trimethylation (H3K27me3) functions as a barrier against through suppression developmental gene expression in PSCs. Here, we have generated human PSC (hPSC) lines which genome-wide reduction H3K27me3 can be induced by ectopic catalytic domain histone demethylase JMJD3 (called JMJD3c). We found that transient, forced...
Abstract During spermatogenesis, meiosis is accompanied by a robust alteration in gene expression and chromatin status. However, it remains elusive how the meiotic transcriptional program established to ensure completion of prophase. Here, we identify protein complex that consists germ-cell-specific zinc-finger ZFP541 its interactor KCTD19 as key regulators mouse prophase progression. Our genetic study shows are co-expressed from pachytene onward play an essential role testis. Furthermore,...
Dry eye disease is the most prevalent pathological condition in aging eyes. One potential therapeutic strategy transplantation of lacrimal glands, generated vitro from pluripotent stem cells such as human embryonic cells, into patients. preceding requirements a method to differentiate gland epithelium cells. As first step for this approach, study aims identify set transcription factors whose overexpression can promote differentiation epithelium-like We performed microarray analyses glands...
Transcription factors (TFs) play a pivotal role in determining cell states, yet our understanding of the causative relationship between TFs and states is limited. Here, we systematically examine state changes human pluripotent embryonic stem cells (hESCs) by large-scale manipulation single TFs. We establish 2,135 hESC lines, representing three clones each 714 doxycycline (Dox)-inducible genes including 481 TFs, obtain 26,998 microscopic images 2,174 transcriptome datasets—RNA sequencing...
The genome of differentiated somatic nuclei is remodeled to a totipotent state when they are transplanted into enucleated oocytes. To clarify the mechanism this remodeling, we analyzed changes in composition core histone variants nuclear-transferred embryos, since recent evidence has revealed that chromatin structure can be as result variant replacement. We found donor cell-derived H3 H3.1, H3.2, and H3.3, well H2A H2A.Z, were rapidly eliminated from Accompanying removal, oocyte-stored H2A.X...
Direct generation of skeletal muscle cells from human pluripotent stem (hPSCs) would be beneficial for drug testing, discovery, and disease modelling in vitro. Here we show a rapid robust method to induce myogenic differentiation hPSCs by introducing mRNA encoding MYOD1 together with siRNA-mediated knockdown POU5F1 (also known as OCT4 or OCT3/4). This integration-free approach generates functional myotubes sarcomere-like structure fusion capacity several days. The silencing facilitates...
Although mouse oocytes progressively acquire meiotic competence during their growth in the ovaries, only half of full-grown can accomplish meiosis. Two types have been reported on basis chromatin configuration, surrounded-nucleolus (SN) type and non-surrounded-nucleolus (NSN) type. Therefore, collected from ovaries adult animals comprise a heterogeneous population; some are meiotically incompetent (NSN-type), competent (SN-type). In present study, we found that could be classified into two...
Genome-wide distribution of the majority H2A and H3 variants (H2A, H2AX, H2AZ, macroH2A, H3.1, H3.2 H3.3) was simultaneously investigated in mouse embryonic stem cells by chromatin immunoprecipitation sequencing. Around transcription start site, histone variant differed between genes possessing promoters high low CpG density, regardless their expression levels. In intergenic regions, regulatory elements were enriched H2A.Z H3.3, whereas repeat abundant respectively. Analysis combinations...
Mouse zinc finger and SCAN domain containing 4 (Zscan4) proteins, which are encoded by multiple copies of Zscan4 genes, expressed specifically in preimplantation embryos vivo embryonic stem (ES) cells vitro. However, the expression patterns mouse have been largely elusive. Here, we show that proteins adult ovaries testes. In ovaries, were detected germinal vesicle (GV) stage oocytes antral follicles, indicating genes activated during diplotene/dictyate meiotic prophase I. Remarkably, showed...
Here we describe a protocol to obtain highly pure cardiomyocytes and neurons from human induced pluripotent stem cells (hiPSCs) via metabolic selection processes. Compared conventional purification protocols, this approach is easier perform scale up more cost-efficient. The can be applied hiPSCs embryonic cells. For complete details on the use execution of protocol, please refer Tohyama et al. (2016) Tanosaki (2020).
Abstract The Zinc finger and SCAN domain containing 4 (ZSCAN4) protein, expressed transiently in pluripotent stem cells, gametes, early embryos, extends telomeres, enhances genome stability, improves karyotypes mouse embryonic (mES) cells. To gain insights into the mechanism of ZSCAN4 function, we identified genome-wide binding sites endogenous protein using ChIP-seq technology human ES where expression was induced by treating cells with retinoic acids or overexpressing DUX4. We revealed...
Transplantation of pancreatic β cells generated in vitro from pluripotent stem (hPSCs) such as embryonic (ESCs) or induced (iPSCs) has been proposed an alternative therapy for diabetes. Though many differentiation protocols have developed this purpose, lentivirus-mediated forced expression transcription factors (TF)—PDX1 and NKX6.1—has at the forefront its relatively fast straightforward approach. However, considering that will be used therapeutic purposes future, it is desirable to develop...
Mouse Zinc finger and SCAN domain containing 4 (Zscan4) is encoded in multiple copies of Zscan4 genes, which are expressed late two-cell stage preimplantation embryos 1–5% the embryonic stem (ES) cell population at a given time. Due to highly identical nucleotide sequences paralogs pseudogenes mouse genomic cluster, previous analyses have been done using exogenous transgenes under regulation Zscan4c promoter. In this manuscript, we generated knock-in ES lines lines, expression endogenous...