ABSTRACT
Although many factors have been identified to be involved in the development of the neuroectoderm during embryogenesis, it is still important to identify novel factors that convert undifferentiated embryonic cells into neuroectoderm. RuvB-like protein 2 (Ruvbl2) is known to regulate gene expression via chromatin remodeling by participating in multi-protein complexes, but its role during embryonic development is not well known. In this study, we established Ruvbl2-overexpressing mouse embryonic stem cells and examined their capacity to specifically differentiate into neuroectoderm and confirmed the specific expression of RUVBL2 in early embryonic neuroectoderm. Our results suggest that Ruvbl2 has a role in the differentiation of neuroectoderm during early embryogenesis.
Subject(s)
Cell Differentiation/genetics , DNA Helicases/metabolism , Mouse Embryonic Stem Cells/cytology , Mouse Embryonic Stem Cells/metabolism , Neural Plate/cytology , ATPases Associated with Diverse Cellular Activities , Animals , Biomarkers/metabolism , Cell Proliferation , DNA Helicases/genetics , Embryoid Bodies/cytology , Embryoid Bodies/metabolism , Fetus/metabolism , Gene Expression Profiling , Gene Expression Regulation, Developmental , Mice , Mice, Inbred C57BL , Organogenesis/genetics , Up-Regulation/geneticsABSTRACT
Spermatogonial stem cells (SSCs) can spontaneously dedifferentiate into embryonic stem cell (ESC)-like cells, which are designated as multipotent SSCs (mSSCs), without ectopic expression of reprogramming factors. Interestingly, SSCs express key pluripotency genes such as Oct4, Sox2, Klf4 and Myc. Therefore, molecular dissection of mSSC reprogramming may provide clues about novel endogenous reprogramming or pluripotency regulatory factors. Our comparative transcriptome analysis of mSSCs and induced pluripotent stem cells (iPSCs) suggests that they have similar pluripotency states but are reprogrammed via different transcriptional pathways. We identified 53 genes as putative pluripotency regulatory factors using an integrated systems biology approach. We demonstrated a selected candidate, Positive cofactor 4 (Pc4), can enhance the efficiency of somatic cell reprogramming by promoting and maintaining transcriptional activity of the key reprograming factors. These results suggest that Pc4 has an important role in inducing spontaneous somatic cell reprogramming via up-regulation of key pluripotency genes.
Subject(s)
Cellular Reprogramming/genetics , DNA-Binding Proteins/genetics , Gene Expression Profiling , Nuclear Proteins/genetics , Transcription Factors/genetics , Adult Stem Cells/cytology , Adult Stem Cells/metabolism , Animals , Blotting, Western , Cells, Cultured , Cluster Analysis , DNA-Binding Proteins/metabolism , Induced Pluripotent Stem Cells/cytology , Induced Pluripotent Stem Cells/metabolism , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/metabolism , Mice , Mice, Inbred C57BL , Mouse Embryonic Stem Cells/cytology , Mouse Embryonic Stem Cells/metabolism , Nuclear Proteins/metabolism , Octamer Transcription Factor-3/genetics , Octamer Transcription Factor-3/metabolism , Oligonucleotide Array Sequence Analysis , Proto-Oncogene Proteins c-myc/genetics , Proto-Oncogene Proteins c-myc/metabolism , Reverse Transcriptase Polymerase Chain Reaction , SOXB1 Transcription Factors/genetics , SOXB1 Transcription Factors/metabolism , Systems Biology/methods , Transcription Factors/metabolismABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Alpinia officinarum Hance (Zingiberaceae) is an annual plant. Its rhizome has long been used as an anti-inflammatory, an analgesic, a stomachic and a carminative in traditional medicine. OBJECTIVE: The aim of this study was to test the anti-inflammatory effects of Alpinia officinarum rhizomes on acute and chronic arthritis in SD rats. METHODS: Alpinia officinarum rhizomes were extracted by refluxing using 80% ethanol. The fractions were prepared by the fractionation of ethyl acetate (EtOAc), n-butanol, and water. This extract was administrated to rats by peroral injection. Acute arthritis was induced by a subcutaneous injection of carrageenan into the hind paw of SD rats. Chronic arthritis was stimulated by a subcutaneous injection of complete Freund's adjuvant (CFA) into the hind paw of SD rats. The paw volume was measured using a plethysmometer, thermal hyperalgesia was tested using a thermal plantar tester, hyperalgesia was evaluated by ankle flexion evoked vocalizations, and the expression of c-Fos in the brain hippocampus was measured with the avidin-biotin-peroxidase technique. Nitric oxide (NO) production was evaluated on nitrite by a Griess assay in lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells. RESULTS: An 80% ethanolic extract showed acute anti-inflammatory activity that it reduced the edema volume in carrageenan-stimulated arthritis and inhibited NO generation in LPS-induced RAW 264.7 cells. In addition, this extract showed chronic anti-rheumatic and analgesic activities by suppressing the swelling volume, by recovering the paw withdrawal latency, and by inhibiting the flexion scores in CFA-induced arthritis. Particularly, this medicine had potent meaningful effects on the second signal of the left hind paw in the form of an immunological reaction compared to its effects on the first signal in the right hind paw after the CFA treatment. This also shows an anti-psychiatric effect through control of the expression of the c-Fos protein of the brain hippocampus in CFA-stimulated arthritis. On the other hand, each fraction showed acute anti-inflammatory effects; the action of the EtOAc fraction may have resulted from the suppression of NO production. CONCLUSIONS: Alpinia officinarum rhizomes may be viable therapeutic or preventive candidates for the treatment of acute and chronic arthritis.
