| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1992: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1991: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
Planarian ! It is an astonishing animal. It can regenerate to a complete body from a lump of only 10^5 cells. We have tried to elucidate the molecular mechanism of the planarian regeneration and clarified the following facts.
1 : Melatonin, which inhibits the fissioning of planarian, inhivited the planarian regeneration. Its' inhibition was specific for melatonin and reversible. Serotonin, N-acetylserotonin and 6-hydroxymelatonin did not inhibit the regeneration. 2 : Melatonin inhibited the regeneration of eye, head and also tail. 3 : By the radio-immunoassary using anti-melatonin antibody, the melatonin content in the head part was higher than that in tail part. 4 : By the Polymerase Chain Reaction (PCR) method, the N-acetyltransferese gene was identified. 5 : Similar proteins (with Mw 65,000 and 62,000) to the frog rhodopsin were identified in the part of head (including eyes) and also auricles. 6 : A heat stable inhibitory-factor for the planarian head-regeneration was isolated from
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the planarian homogenate. the fractionation by the ammonium sulfate precipitation, by the solubilization with detergent and by the DEAE Sephadex column chromatography were usefull in the purification. though this factor (with the Mw larger than 10,000) inhibited the regeneration of eye and head, it did not that of the tail. 7 : On the assumption of that the decapitation accompanies a great stress like the heat treatment, the heat shock genes, HSP90 and HSP60, were analyzed in the planarian. Nucleotide sequence for the genes of HSP90 and HSP70 were partially determined. A transient (18-20 hrs after the decapitation) expression of HSP90 mRNA was detected. 8 : Several monoclonal antibodies for the organs, such as testis and gland cells, were obtained. 9 : The culture of planarian cells was unsuccessful because of its fragile cells membrane. The culture of A375 cells at the interface of the culture medium and fluorocarbon (FC-70) was carried out. The trypsintreatment in the recovery of cells was unnecessary and the cells with intact membrane proteins were obtained. Less
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