Takesue Sachiko, Onitake Kazuo, Keino Hiroomi, Takesue Yoshiki
Biological Institute, Faculty of Sciences, Nagoya University, Chikusa-ku, 464, Nagoya, Japan.
Institute for Developmental Research, Aichi Prefectural Colony, 480-03, Kasugai, Aichi, Japan.
Wilehm Roux Arch Dev Biol. 1983 May;192(3-4):113-119. doi: 10.1007/BF00848679.
Vitellin was purified from eggs of the silkworm,Bombyx mori, by a new method in which vitellin was extracted from isolated yolk granules. The purified vitellin had a molecular weight of 540,000. An antibody against purified vitellin was prepared in rabbits. It reacted with the hemolymph vitellogenin as well as with purified vitellin, but not with other proteins in the hemolymph or in the extract from yolk granules. The anti-vitellin IgG was used to immunocytochemically locate vitellin in theBombyx non-diapause egg during early developmental stages. In the egg, just after oviposition, vitellin was located in internal yolk granules and in small yolk granules of the periplasm. During the early developmental stages studied, vitellin was not metabolized uniformly throughout the egg. The vitellin of the internal yolk granules located at the posterior-dorsal part and of the small peripheral yolk granules was utilized in 16 h and 2 days, respectively, after oviposition. A thin, very vitellin-poor layer was located between the periplasm and the vitellin-rich interior in the newly laid egg. it was always in close contact with the periphery where blastoderm and germ-band cells developed.
采用一种从分离的卵黄颗粒中提取卵黄磷蛋白的新方法,从家蚕卵中纯化出了卵黄磷蛋白。纯化后的卵黄磷蛋白分子量为540,000。用纯化的卵黄磷蛋白对家兔进行免疫,制备了抗卵黄磷蛋白抗体。该抗体能与血淋巴中的卵黄原蛋白以及纯化的卵黄磷蛋白发生反应,但不与血淋巴或卵黄颗粒提取物中的其他蛋白质发生反应。利用抗卵黄磷蛋白IgG对处于早期发育阶段的家蚕非滞育卵中的卵黄磷蛋白进行免疫细胞化学定位。在卵刚产下后,卵黄磷蛋白位于内部卵黄颗粒和周质中的小卵黄颗粒中。在所研究的早期发育阶段,整个卵中的卵黄磷蛋白并非均匀地被代谢。位于后背部的内部卵黄颗粒和外周小卵黄颗粒中的卵黄磷蛋白分别在产卵后16小时和2天被利用。在新产下的卵中,周质和富含卵黄磷蛋白的内部之间有一层很薄且卵黄磷蛋白含量极低的层。它始终与胚盘和胚带细胞发育的周边紧密接触。