Insect eggs store many lipid droplets as an energy source for embryonic development. We previously reported that lipid droplets are incorporated into embryos in three steps in the silkworm, Bombyx mori. The midgut plays important roles in lipid incorporation during the second and third steps, whereas the manner of lipid incorporation during the first step is still unknown. In this study, we focused on how lipids were incorporated into the embryo in the first step, compared with the mechanisms used in the second step, by means of transmission electron microscopy using the high-pressure freezing and freeze substitution method. At the beginning of the first step (blastoderm formation stage), some lipid droplets were observed in each cell of the embryonic tissues. Lipid droplets were seen to be derived from the oocyte peripheral cytoplasm by superficial cleavage. At the end of the first step (late appendage formation stage), some lipid droplets were attached to the elongated rough endoplasmic reticulum (rER). It seemed that formation of the lipid droplets occurred in embryonic cells at the end of the first step, because the rER is the site of biogenesis of lipid droplets. The incorporation of lipid droplets in the first step may be subdivided into two stages: the blastoderm formation stage and the subsequent stage before blastokinesis.
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