| Project/Area Number |
07507007
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Embryonic/Neonatal medicine
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| Research Institution | Shimane Medical University |
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Principal Investigator |
OTANI Hiroki Shimane Medical University, Department of Anatomy, Professor, 医学部, 教授 (20160533)
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| Co-Investigator(Kenkyū-buntansha) |
TATEWAKI Reiko Shimane Medical University, Department of Biology, Instructor, 医学部, 助手 (10112129)
NAORA Hiroyuki Shimane Women's College, Department of Food Science, Instructor, 家政科, 助手 (70222156)
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)
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| Project Period (FY) |
1995 – 1997
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| Keywords | exo utero development / gene introduction / liposome / electroporation / antisense oligonucleotide / histogenesis |
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| Research Abstract |
Exo utero development system and techniques of microinjection into fetuses have been improved. To increase efficiency of the uptake of exogenous genes injected into mouse embyonic organs, optimal conditions of gene carriers and electroporation have been established, and usefulness of antisense oligonucleotides has been estimated. To demonstrate the potential of exo utero system, several biologically active substances were injected into the specific fetal organs, and their effects were analyzed. The exo utero experimental systems to analyze the histogenesis of several organs have been established.
1. Improvement of exo utero system : Reduction of manipulation time for each procedure, modification of incision of the uterine wall, anesthesia and resuscitation, all together have significantly improved the survival rate after the manipulation and efficiency of microinjection.
2. Optimization of conditions for gene introduction : Among several liposomes, the ones with the highest efficiency of
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reporter gene incorporation under different conditions were determined. Conditions of electroporation with the BTX system were evaluated and the flat type electrode showed an excellent handling. Conditions have been established for good diffusion of injected solution into the cerebral parenchyme with minimum damage to the tissue.
3. Introduction of antisense oligonucleotides : Antisense oligonucleotides for Tag1 and Mab21 was injected into the fetal cerebral ventricle at single-dose or double-dose with an interval, and the protein expression levels decreased significantly.
4. Application of exo utero system into the functional analysis of regulatory molecules in the histogenesis of the organs : LIF was injected into the cerebral ventricle and showed specific effects on the cerebral cortex including increase in the neuroblasts and fibers in the specific cortical layrs. The effects of ACTH on the fetal adrenal were similarly evidenced by the established exo utero experimental system. The exo utero experimental systems for analysis of pathogenesis of hydrocephaly and excencephaly, and development of the inner ear and joints have also been established. Less
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