| Project/Area Number |
62570234
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hygiene
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| Research Institution | The Jikei University School of Medicine |
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Principal Investigator |
MATSUMOTO Nobuo The Jikei University School of Medicine, 医学部, 教授 (30009998)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIBA Shigeo The Jikei University School of Medicine (90056549)
小野澤 照夫 東京慈恵会医科大学, 医学部, 助手 (80152544)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | acridine orange / fluorescence microscope / vital staining / digital malformation / マウス胚 / 発生毒性 / 生体染色法 / 器官形成期胎芽 / アクリジン・オレンジ(AO) |
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| Research Abstract |
In this study, normal and abnormal growth of post-implantation embryos was investigated by means of vital staining with acridine orange using fluorescence microscope. Embryos from mother mice injected with AO, emitted intense secondary fluorescence five minutes after injection and still did emit weak secondary one 24 hrs. after. But 48 hrs. after, embryos did illuminate only auto-fluorescence without the secondary one.
Pregnant mice were administered with Mitomycin C and Caffeine simultaneously on 9th day of gestation. This treatment has been well known to cause digital malformation of the embryo at a high incidence. Then, on the successive 10th and 11th day of gestation, one hr. after AO injection into mother mice, embryos were taken out of uterine horns and investigated by fluorescence microscope.
From the time course investigation, we confirmed that the secondary fluorescence existed longer in damaged tissue cells of embryos by treatment with Mitomycin C and caffeine than in control ones, as living embryonic cell can excrete AO while damaged cells can not.
There were many granule-like cell masses with intense yellowish-green fluorescence diffusely on the whole body of embryo and those specifically existed with high density at the mesenchymal tissue of limb buds without formation of digital rays.
From the results, above mentioned, normal and abnormal embryonic growth can be investigated in detail by color and intensity of fluorescence using fluorescence microscopy through AO vital staining.
Further investigation in combination with whole embryo culture is expected to bring us a fruitful result about normal and abnormal embryonic growth.
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