Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Rhabdomere Formation in Late Pupal Stage of Drosophila melanogaster; Observation Using High-Pressure Freezing and Freeze-Substitution, and High-Voltage Electron Microscopy

초고압 동결장비와 초고압투과전자현미경을 이용한 초파리의 감간분체 형성과정의 구조분석

  • Mun, Ji-Young (Laboratory of Cell Engineering & 3-D Structure, Graduate School of Life Sciences and Biotechnology, Korea University) ;
  • Arii, Tatsuo (Section of Brain Structure, Center for Brain Experiment, National Institute for Physiological Sciences, Okazaki National Research Institutes) ;
  • Hama, Kiyoshi (Section of Brain Structure, Center for Brain Experiment, National Institute for Physiological Sciences, Okazaki National Research Institutes) ;
  • Han, Sung-Sik (Laboratory of Cell Engineering & 3-D Structure, Graduate School of Life Sciences and Biotechnology, Korea University)
  • 문지영 (고러대학교 생명과학대학 생명공학원 세포공학 및 생체3차구조 연구실) ;
  • ;
  • ;
  • 한성식 (고러대학교 생명과학대학 생명공학원 세포공학 및 생체3차구조 연구실)
  • Published : 2007.03.30
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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 rhabdomere could be observed in the cytoplasm of the photoreceptor rolls, which were visible during the profess 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.

감간분체 (Rhabdomere)가 왕성하게 형성되는 번데기시기의 Drosophila melanogaster 광수용체 세포를 고정효과가 가장 좋은 high-pressure freezing (HPF)와 freeze-substitution (FS) 기법으로 고정한 후, 초고압 전자현미경을 통하여 1000 KV 전압에서 관찰하였다. 후박절편(250nm)의 tilting image를 통하여 광수용체 세포에서 감간분체 형성의 세가지 형태가 관찰되었다. 첫째로, 감간분체의 육방정계 배열이 (hexagonal may) 서로 다른 각도로 존재하고 있음을 관찰할 수 있었다. 둘째, 광수용체 세포의 세포질 안에서 감간분체 형성의 중간 과정으로 보여지는 작은 조각의 감간분체를 관찰할 수 있었다. 셋째, 감각분체가 여러 개의 층으로 형성되어 있음을 관찰할 수 있었다. 또 광수용체 세포의 세포질에는 최소한 세 가지의 소낭(vesicle)이 존재하였고, 이들 소낭들은 번데기 시기의 감각분체 형성을 주도하는 것으로 확인되었다. 이 결과들에 의하여 소낭이 감각분체 형성과 감각분체의 부분들이 모여서 감각분체를 완성되는 과정에 관여함을 추정할 수 있었다.

Keywords

References

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