Kohtaro Shibahara's research while affiliated with Kyushu University and other places

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Publications (1)


FIGURE 1. Exencephaly in YB-1 / embryos. A, PCR genotyping of yolk sac DNA from nine E11. 5 embryos. M, size marker. Bands of 650 bp of wild-type (WT) and 865 bp of mutant (disrupted) are shown. Total embryo protein (E11.5) was isolated, and the amount of YB-1 protein was determined by Western blotting using a polyclonal YB-1 antibody (lower panel). B, YB-1 / and YB-1 / embryos at E11.5 to E14.5 stages of development. Exencephaly was observed in various embryonic stages of YB-1 null mice. Arrow indicates brain tissue outside of the calvarium in exencephalic embryos. C, YB-1 / and YB-1 / embryos at E10.5 stage of development. YB-1 / embryos show severe hemorrhage (bottom left panel) and anemia (bottom right panel) in comparison with wild-type embryos (top panels).
FIGURE 2. YB-1 expression in embryonic tissues. A, nonradioactive in situ hybridization of wild-type E13.5 mouse embryos with antisense or sense YB-1specific probes shows that YB-1 is expressed in the whole embryo region. Sense, negative control; H&E, hematoxylin and eosin staining. Scale bar 2 mm. B, in situ hybridization of wild-type E13.5 embryos showing expression of YB-1 in mouse organs. Left panel, scale bar 100 m. Right panel, scale bar 20 m. C, immunohistochemistry showing YB-1 expression in wild-type and YB-1 null E13.5 embryos. No staining was observed in the ganglion of the YB-1 / mouse. Scale bar 100 m.
FIGURE 4. Elevated EF-1 expression in YB-1 / embryos and decreased growth of YB-1 / MEFs. A, Western blot analysis of protein expression in E11.5 embryonic mouse tissues. Total protein derived from eight PCR-genotyped mouse embryo tissues (50 g of protein per lane) was immunoblotted using a specific antibody against YB-1, EF-1, p70 S6K, eIF4E, Akt, and PCNA. Elevated levels of EF-1 were observed in YB-1 / MEFs (lanes 3 and 5). Relative band intensity (%) is presented. B, establishment of MEFs. Western blot analysis of YB-1 and PCNA expression levels after establishment of immortalized, PCR-genotyped MEF clones (left panel) and immortalized YB-1 null MEF clones transfected with a pIRES (control) vector or pIRES-YB-1 plasmid (right panel). C, growth curves of YB-1 / , YB-1 / , and YB-1 / MEFs. One representative experiment is shown. Population doubling curves were determined using trypan blue exclusion. D, proliferation rate of MEFs as assessed by cell counts. YB-1 / (diamonds), YB-1 / (squares), and YB-1 / (triangles) were inoculated at 5 10 4 cells/ml. The cell numbers were determined at the time points indicated. Ectopic expression of wild-type YB-1 reversed the proliferation defect (right panel).
FIGURE 5. Actin expression in the brain of wild-type and mutant embryos. A, at low magnification, ␤ -actin was shown to be ubiquitously expressed in wild-type embryos; however, YB-1 Ϫ / Ϫ embryos demonstrated local reduc- 
FIGURE 6. Colony transformation activity was reduced in YB-1 / MEFs but could be rescued by re-expression of YB-1. A, three YB-1 / MEF cell lines (middle panel) demonstrated reduced transformation activity compared with wild-type MEFs (top panel), following 2% Giemsa staining. Introduction of recombinant YB-1 restored the transformation activity (bottom panel). B, three YB-1 / MEF cell lines demonstrated reduced colony forming activity compared with wild-type MEFs, following 2% crystal violet staining. Cells were assayed in triplicate.

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YB-1 is important for an early stage embryonic development - Neural tube formation and cell proliferation
  • Article
  • Full-text available

January 2007

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150 Reads

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110 Citations

Journal of Biological Chemistry

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Abbas Fotovati

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Takakazu Sasaguri

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The eukaryotic Y-box-binding protein-1 (YB-1) is involved in the transcriptional and translational control of many biological processes, including cell proliferation. In clinical studies, the cellular level of YB-1 closely correlates with tumor growth and prognosis. To understand the role of YB-1 in vivo, especially in the developmental process, we generated YB-1 knock-out mice, which are embryonic lethal and exhibit exencephaly associated with abnormal patterns of cell proliferation within the neuroepithelium. β-Actin expression and F-actin formation were reduced in the YB-1 null embryo and YB-1-/- mouse embryonic fibroblasts, suggesting that the neural tube defect is caused by abnormal cell morphology and actin assembly within the neuroepithelium. Fibroblasts derived from YB-1-/- embryos demonstrated reduced growth and cell density. A colony formation assay showed that YB-1-/- mouse embryonic fibroblasts failed to undergo morphological transformation and remained contact-inhibited in culture. These results demonstrate that YB-1 is involved in early mouse development, including neural tube closure and cell proliferation.

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Citations (1)


... To further confirm this, we have initiated an experiment to knock out the ybx1 gene from the zebrafish genome, which promises to provide insights into the functional relationship of Ybx1 with its counterpart YB-1 and germ cell-specific YB-2 in the tetrapods. The knockout of YB-1 in the mouse causes embryonic lethality due to its ubiquitous expression and universal functions in somatic cells [40]. In contrast, the disruption of YB-2 gene in the mouse genome leads to infertility without any effect on vitality [41]. ...

Reference:

Proteomic analysis of zebrafish Folliculogenesis identifies YB-1 (Ybx1/ybx1) as a potential gatekeeping molecule controlling early ovarian Folliculogenesis
YB-1 is important for an early stage embryonic development - Neural tube formation and cell proliferation

Journal of Biological Chemistry