ABSTRACT: During mammalian spermatogenesis the mouse VASA homolog (MVH), a germ cell-specific DEAD-box type RNA binding protein, localizes in a germline-specific RNA granule termed the chromatoid body (CB). Genetic analyses have revealed that MVH is essential to progress through spermatogenesis, although the molecular mechanisms of its function remain elusive. We found that the acetyltransferase HAT1 and its cofactor, p46, are specifically co-localized with MVH in the CB and acetylate MVH at Lys-405, leading to inactivation of its RNA binding activity. Notably, the acetylation is developmentally regulated, paralleling the temporally regulated co-localization of HAT1 and p46 in the CB. We have identified 858 mRNAs as MVH targets, a large proportion of which correspond to previously identified translationally arrested genes. Importantly, eIF4B mRNA, a target of MVH, is selectively released from MVH-RNP when MVH is acetylated, paralleling an increase in eIF4B protein. These findings reveal a novel signaling pathway that links acetylation to RNA processing in the control of spermatogenesis. IMVH target mRNA was purified from MVH immuno precipitated complex, followed by RNA purification and identified by DNA chip. In this study, we identified more than 800 MVH target mRNA in testis.
Project description:Differences of metabolism-related gene expression profiles in human ESCs and ESC-derived purified cardiomyocytes were analyzed and successfully identified. Human ESCs and ESC-derived purified cardiomyocytes were used for this experiment.
Project description:We newly identified skeletal muscle differentiation-associated miRNAs by comparing miRNA expression profile between C2C12 cell and Wnt4-overexpressing C2C12 cell. miR-487b, miR-3963 and miR-6412 are significantly down-regulated in differentiating C2C12 cells, and transfection of their mimics resulted in reduced expression of myogenic differentiation markers including Troponin T, myosin heavy chain fast and slow type. Single analysis for each condition (proliferating C2C12 cells, differentiating C2C12 cells, proliferating Wnt4-overexpressing C2C12 subline cells
Project description:Comprehensive gene expression analysis in BM-resident stromal cells was performed for an overview of BM environmental change caused by total body irradiation (TBI). Total RNA samples collected from BM-resident stromal cells with or without TBI were subjected to high sensitivity DNA microarray assays Three-condition experiment: Unirradiated, 1 day after TBI and 3 days after TBI. Bone marrow stromal cells were obtained from C57BL/6 mice (n = 6) either non-irradiated or after 9.5 Gy irradiation at indicated times.
Project description:The change of mRNA expression in murine immortalized podocyte were analyzed after miR-26a silencing. These results provide a basical information of molecular pathology in podocyte biology. Mouse podocytes immortalized by temperature sensitive SV40 were used. Podocyte cultures grown at 33 °C were trypsinized and then cultured with RPMI-1640 without antibiotics in 24-well plates at 60–70% confluence for 2 days. On day 3, an anti-miR negative control (40 pmol) or the miR-26a miRNA inhibitor (40 pmol) was transfected to podocytes. The cells were analyzed after culturing for 24 hour.
Project description:Transcriptional profiling of left ventricular tissues of Dahl rat with or without treatment of chaetocin Three-condition experiment, Control vs. failing heart, failing heart vs. treatment with chaetocin. 3 samples mixture per each group
Project description:We previously demonstrated that hematopoietic stem cell (HSC)-like cells are robustly expanded from mouse embryonic stem (ES) cells by enforced expression of Lhx2, a LIM-homeobox domain (LIM-HD) transcription factor. Here we established an ES cell line which conditionally expressed Lhx2 by Tet-On system. The ES cells were differentiated into HSC-like cells by Lhx2 expression. Lhx2-regulated genes were identified by comparing the HSC-like cells with those cultured in the absence of Lhx2 expression. Mouse ES with inducible Lhx2 were differentiated on OP9 stromal cells into hematopoietic lineage. On day 5 of the differentiation induction,Lhx2 expression was started by the addition of doxycycline (dox) and the cells were cultured on OP9 stromal cells in the presence of IL-6 and SCF. On day 20, HSC-like cells were harvested and re-seeded onto OP9 stromal cells in the absence of Lhx2 expression by dox-removal for 3 days. these cells were compared with the original HSC-like cells.
Project description:Recent studies have demonstrated that micro (mi)RNA molecules can be detected in the circulation and can serve as potential biomarkers of various diseases. This study used microarray analysis to identify aberrantly expressed circulating miRNAs in patients with type 1 autoimmune hepatitis (AIH) compared with healthy controls. Patients with well-documented and untreated AIH were selected from the National Hospital Organization (NHO)-AIH-liver-network database. They underwent blood sampling and liver biopsy with inflammation grading and fibrosis staging before receiving treatment. To further confirm the microarray data, circulating expression levels of miR-21 and miR-122 were quantified by real-time quantitative polymerase chain reaction in 46 AIH patients, 40 patients with chronic hepatitis C (CHC), and 15 healthy controls. Consistent with the microarray data, serum levels of miR-21 were significantly elevated in AIH patients compared with CHC patients and healthy controls. miR-21 and miR-122 serum levels correlated with alanine aminotransferase levels. Circulating levels of miR-21 and miR-122 were significantly reduced in AIH patients with liver cirrhosis, and were inversely correlated with increased stages of fibrosis. By contrast, levels of circulating miR-21 showed a significant correlation with the histological grades of inflammation in AIH. We postulate that aberrantly expressed serum miRNAs are potential biomarkers of AIH and could be implicated in AIH pathogenesis. Alternations of miR-21 and miR-122 serum levels could reflect their putative roles in the mediation of inflammatory processes in AIH. Case-control study, steroid treatment
Project description:We examined the metastasis-related miRNAs induced by SDF-1/CXCR4 system, using oral cancer cells. Consequently, we identified 4 kinds of upregulated-miRNAs in B88-SDF-1. The metastasis-related miRNAs induced by SDF-1/CXCR4 system in oral cancer are largely unknown. Thus, we examined the metastasis-related miRNAs induced by SDF-1/CXCR4 system, using four types of oral cancer cells; mock cells vs forced-expression of SDF-1cells and parental cells vs parental cells treated by SDF-1.
Project description:To identify molecular biomarkers that are useful for diagnosis and its targeting treatment, we analysed expression profile of synovial sarcoma tissue. In the present study, we studied gene expression profiles comparing 11 cases of synovial sarcoma.