| Co-Investigator(Kenkyū-buntansha) |
HASHIMOTO Ryuju Shimane Medical University, Department of Anatomy, Instructor, 医学部, 助手 (90252907)
HATTA Toshihisa Shimane Medical University, Department of Anatomy, Instructor, 医学部, 助手 (20238025)
OTANI Hiroki Shimane Medical University, Department of Anatomy, Associate Professor, 医学部, 助教授 (20160533)
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| Research Abstract |
1. Development of serial culture system for mouse whole embryos : (1)Culture system from one cell stage has been improved to culture embryos, including those of closed colonies such as ICR and NOD mice, which were previously unable to be successfully cultured, to the blastocyst stage stably at a high frequency. (2)Culture system for postimplantation period has been devised. Among conditions tried, a system, in which placental cells were used as feeder cells, embryos were separated from feeder cells by Millicell with pored membrane, and high and changing concentrations of fetal bovine serum was supplemented to the medium, gave the higher frequency of embryos developed to the primitive streak stage. (3)The "exo utero"procedure has been improved to observe and manipulate embryos on day 11 of gestation onward and to allow a significantly higher rate of survival of manipulated embryos than reported.
2. Applications of culture systems for embryo manipulations. (1)Several transgenic mouse line
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s with developmental abnormalities, which were created by manipulation of preimplantation stage embryos, were analyzed genetically, morphologically, and developmentally. In a line with hemifacial microsomia, insertional mutation locus (Hfm : hemifacial microsomia-associated locus)due to integration of the transgene to the genome has been mapped and analyzed. Regions conserved among multiple mammlian species have been confirmed at the mutation locus. (2)Using mRNA injection and other methods on Xenopus laevis embryos, developmental events during preorganogenetic period were analyzed. Interaction between growth factors and transcriptional factors was elucidated in body axis determination, mesoderm induction, induction of muscle and nerve tissues. (3)To analyze the role of cell adhesion molecules in the histogenesis of the forebrain cortex at the later gestational period, RGD peptide was injected into the cerebral ventricle by exo utero procedure. As a result, proliferation and migration of neuroblasts were inhibited, suggesting the involvement of cell adhesion molecules. Less
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