Rhabdomere formation in late pupal stage of Drosophila melanogaster; Observation using high-pressure freezing and freeze-substitutiion, and high-voltage electron microscopy
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The late pupal stage of Drosophila melanogaster occurs immediately before the completion of retinal development, during which the rhabdomere rapidly forms. In this period, the photoreceptor cells were fixed and dehydrated using a high-pressure freezer (HPF) and freeze substitution (FS) technique, which is the most effective in preserving the cell structures, and observed using high-voltage electron microscopy (HVEM) at 1000 KV. The rhabdomere was classified structurally into three types of formation patterns using stereo-tiling image of thick sections. Initially, hexagonal arrays of rhabdomere existed in different angles. In addition, small pieces of rhabdomeres could be observed in the cytoplasm of the photoreceptor cells, which were visible during the process of rhabdomere formation. In addition, multiple layers of rhabdomere strings were observed. We observed there are at least three types of vesicles related to rhabdomere formation in photoreceptor cells. In addition, it was found that these vesicles initiate the formation of the rhabdomeres during the pupal stage. Collectively, these data suggest that rhabdomeres were mainly formed through vesicles, and that parts of the rhabdomere formed first and then gathered and formed rhabdomeres in the late pupal stage.
Abstract
The late pupal stage of Drosophila melanogaster occurs immediately before the completion of retinal development, during which the rhabdomere rapidly forms. In this period, the photoreceptor cells were fixed and dehydrated using a high-pressure freezer (HPF) and freeze substitution (FS) technique, which is the most effective in preserving the cell structures, and observed using high-voltage electron microscopy (HVEM) at 1000 KV. The rhabdomere was classified structurally into three types of formation patterns using stereo-tiling image of thick sections. Initially, hexagonal arrays of rhabdomere existed in different angles. In addition, small pieces of rhabdomeres could be observed in the cytoplasm of the photoreceptor cells, which were visible during the process of rhabdomere formation. In addition, multiple layers of rhabdomere strings were observed. We observed there are at least three types of vesicles related to rhabdomere formation in photoreceptor cells. In addition, it was found that these vesicles initiate the formation of the rhabdomeres during the pupal stage. Collectively, these data suggest that rhabdomeres were mainly formed through vesicles, and that parts of the rhabdomere formed first and then gathered and formed rhabdomeres in the late pupal stage.
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- A, "Figure LegendsFig 1. The retinal cell in the late pupal stage of Drosophila melanigaster using high-pressure freezing and freeze-substitution method. A total of 60 images were taken by tilting a thick-sectioned ribbon by 2. Images presented in this figure were 6 apart. Size bar = 1000 nm Fig 2. More information can be obtained by observing a thick section from a part of the rhabdomere formation in progress. A large number of the vesicles are observed in parts of rhabdomere formation and some of them are fused into the rhabdomere being formed . There was an intermediate rhabdomere structure fused into cell membrane in cytoplasm . Two direction of row was observed in rhabdomere stereo-image. Vesicles fused into rhabdomere were observed 3-dimensionally. Size bar = 500 nmFig 4. Variable types of vesicle", double-membra(us), pp.3~