Studies on Developmental Biology of Cultured Antheraea yamamai Embryo

方法：采用器官控制结扎法和胚胎培养法，从胚胎早期到产卵后100天，分阶段观察了1龄幼虫对滞育解除剂、咪唑类衍生物和20-羟基蜕皮激素的反应。与上述研究分开，作为对天蚕育种的基础研究贡献，还研究了天蚕和柞蚕蛋白质的生化种特异性。结果表明：1）结扎实验结果证实了RF和MF之间的拮抗作用以及RF咪唑衍生物的可断裂性对滞育的调节作用。另一方面，鉴于蜕皮激素对天蚕早期幼虫生长的抑制作用，推测胸腺也参与了滞育过程。蜕皮激素对早期蚤状体1龄幼虫的抑制作用不明显， ...更多信息 对75~100日龄幼虫有较强的毒力。根据浸渍法的蜕皮激素的皮肤渗透性，这在处理昆虫激素方面成为问题，作为随访的结果，在浸渍后随着时间的推移观察到其摄取体积的增加。2）免疫学鉴定了Antheraea arylphorin（M.W.83000）和雌性特异性储存蛋白（M.W.75000）。我们发现了一个36 kDa的山脉曲霉肽和34 kDa的佩尼曲霉特异性肽。在杂种的血淋巴中检测到这两种蛋白。我们在脂肪体中发现了一种新的女性特异性蛋白质，并将其暂命名为24K肽（24K）。通过凝胶渗透色谱法测定天然24K蛋白的分子量为43000。血淋巴和卵巢中未检测到~（24）K。因此，24K不应被归类于卵黄蛋白原。对24 K蛋白的氨基酸组成和N端氨基酸序列进行分析，发现24 K蛋白与30 K的小卵黄蛋白原和蝗虫21 kDa雌性特异蛋白没有同源性。采用柱层析法对柞蚕卵黄蛋白进行了纯化。柞蚕卵黄蛋白原由一个大亚基（M.W.21000）组成，而卵黄蛋白由大亚基（M.W.17000）和小亚基（M.W.45000）组成，表明蛋白质裂解发生在内化到卵泡细胞中。山脉圆线虫卵黄蛋白原除卵黄蛋白外，还含有大亚基（分子量185000）。山麦按蚊卵黄蛋白与柞蚕卵黄蛋白之间存在免疫学联系。少

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 … More 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