1993 Fiscal Year Final Research Report Summary
Studies on Developmental Biology of Cultured Antheraea yamamai Embryo
| Project/Area Number |
04454064
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
蚕糸学
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| Research Institution | Shinshu University |
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Principal Investigator |
KOBAYASHI Masaru Faculty of Textile Science and Technology, Shinshu University, 繊維学部, 助手 (10021164)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEI Ryuzo Faculty of Textile Science and Technology, Shinshu University, 繊維学部, 教授 (80021161)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Pharate 1st-instar diapause / Body ligation / Insect hormone / Radioimmunoassay / Allied species / Female specificpritein / Vitellogenin / Vitellin |
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| Research Abstract |
Methodologically, using organ-controlling ligature and cultured embryo, pharate 1st-instar larvae from early embryo to 100 day after ovipoition were examined by stage of growth for their responses to diapause breaking agent, imidazole derivatives and 20-hydroxyecdysone. Separately from the above examination, and as a basic study contribution to the breeding of Antheraea yamamai, allied species of Antheraea was also examined for biochemical species-specificity in proteins of A.yamamai and A.pernyi. The following results were obtained ;
1) The results of ligature experiment confirmed the regulability of diapause by the antagonism between RF and MF and the breakability of RF imidazole derivatives. On the other hand, in view of the supressive effect of ecdysteroid on the growth of early pharate 1st-instar larvae of Antheraea yamamai, thoracic gland was also suggested to be involved i diapause. No such suppressive effect of ecdysteroid as that on early pharate 1st-instar larvae proved not to
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be had on the larvae from 75 day to 100 day. In terms of that skin permeability of ecdysone according to the dipping method, which becomes problematic with respect to the handling of insect hormone, an increase in its uptake volume was observed with the course of time following dipping, as a result of the follow-up.
2) Antheraea arylphorin (M.W.83000) and female specific storage protein (M.W.75000) were identified immunologicaly. We found a 36 kDa A.yamamai peptide and 34 kDa A.pernyi-specific peptide. The two proteins were detected in the hemolymph of hybrids. We discovered a new female specific protein in the fat bodies and called the protein 24K peptide (24K) tentatively. The molecular weight of the native 24K protein was determined 43000 by gel permiation chromatography. 24K was not detected in the hemolymph and ovaries. We conclude that 24K should not be classified in vitellogenin. We analyzed amino acid composition and N-terminal amino acid sequence of 24K.There was no homology among 24K Bombyx LSP and 30K Manduca microvitellogenin, and Locusta 21kDa female specific protein. We purified A.pernyi vitellin by coloumn chromatographies. A.pernyi vitellogenin composes of one large subunit (M.W.21000) but vitellin composes of large (M.W.17000) and small (M.W.45000) subunit, suggesting that protein cleavage occurred at internalization into follicle cells. A.yamamai vitellogenin composes of large subunit (M.W.185000) as well as vitellin. There is immunological relationship between A.yamamai vitellin and A.pernyi one. Less
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