カイコ胚細胞ライソソームのアクリジン・オレンジ取込能の休眠状態による差異
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Several biochemical, physiological, and morphological differences between the diapausing and the developing eggs have been described in the Bombyx silkworm (Chino, 1957a, b, 1958, 1960 and 1961; Okada, 1967 and 1970; Takami, 1957 and 1959). In addition to those differences, the embryos in different conditions of the diapause showed different aspects under a fluorescent microscope when they were vitally stained with acridine orange. Diapause eggs, non-diapause eggs, and artificially evoked non-diapause eggs of two different reces of the Bombyx mori were tested. The embryos were dissected out of the egg shells and were immersed in 0.01% solution of acridine orange in a physiological saline (Takami, 1957) for 10 minutes at a room temperature. They were washed with three changes of the saline and squashed on a slide glass. Leitz fluorescence outfit with the Ortholux microscope was used for the observation with an excitor filter UG 1. Round cytoplasmic particles with flaming orange fluorescence were observed in embryo cells. These were two types of the particles, large (2-3μ in diameter) and small (0.8-1μ in diameter). Pre-diapause embryos and post-diapause embryos, the diapause of which had been evoked by a cold treatment, included many large particles and small number of the small ones, and developing embryo cells were loaded with the small particles exclusively. None of the cytoplasmic particles were observed in diapausing embryo cells to emit flaming orange fluorescence. Chilling the diapausing egg at 5℃ for 2 weeks a few particles in each cell of the embryo recovered their stainability with acridine orange. Although the cold treatment for as long as a month induced much larger number of particles to stain, it took further 2 months to get to the maximum in the number of the particles to stain with acridine orange. The embryo did not terminate the diapause until it had been chilled for more than 3 months at 5℃. Correlation between the pattern of acridine orange-stainable particles in the cells of an embryo and the intensity of the diapause of the embryo seems so close that the latter probably be estimated by investigating the former. Those particles showing flaming orange fluorescence in the silkworm embryo cells vitally stained with acridine orange were identified as lysosomes by comparing them with particles exhibiting acid phosphatase activity, for their shape, size, number and topographical distribution in the cytoplasm. Besides, those particles resembled lysosomes as observed under an electron microscope (Okada, 1970) in their size and time of appearance with the small ones. This may be further supported by that cytoplasmic inclusions which stain vitally with acridine orange have been established to be lysosomes in rat muscle cells (Canonico et al., 1969), liver cells (Zelenin, 1966), brain cells and kidney cells (Koenig, 1963a and b). The lysosomes, i. e., acid phosphatase-containing particles, were cytochemically found to exist even in the diapausing embryo cells, although they did not stain with acridine orange in those embryo cells. The lack of stainability of the lysosomes with the dye during the diapause seems to suggest that properties of lysosomal mambrane might change during the process of the diapause.
- 社団法人日本動物学会の論文
- 1970-07-15
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