• 제목/요약/키워드: Ultrastructural change

검색결과 82건 처리시간 0.019초

고압전자현미경을 이용한 소뇌 평행섬유-조롱박세포간 신경연접의 3차원 재구성 (3-Dimensional Reconstruction of Parallel fiber-Purkinje Cell Synapses Using High-Voltage Electron Microscopy)

  • 이계주;권희석;강지선;유임주
    • Applied Microscopy
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    • 제35권1호
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    • pp.31-39
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    • 2005
  • 신경연접은 신경세포 사이의 신호전달을 위해 형성되는 미세구조로 다양한 생리적, 병리적 상태에 반응하여 형태적, 기능적 변화를 보인다. 현재까지 투과전자현미경을 이용한 신경연접 미세구조의 2차원적 연구들이 많은 유용한 정보를 제공하여 왔으나 신경연접 구성요소들을 보다 정확하게 분석하고 전신경연접부위와 후신경연접부위의 정확한 연결관계를 이해하기 위해서는 신경연접의 3차원 재구성이 요구된다. 고압전자현미경은 고해상도와 시료투과력의 증가로 인해 두꺼운 절편의 관찰이 가능하며 이를 통해 미세구조의 3차원적 특성을 규명하는 것이 용이하므로, 신경연접의 3차원 재구성에 고압전자현미경을 응용하는 것은 많은 수의 연속절편 제작과 오랜 기간의 영상처리가 요구되는 기존의 재구성 방법의 난점들을 극복할 수 있을 것으로 생각된다. 이에 본 연구에서는 고압전자현미경을 이용하여 흰쥐 소뇌 평행섬유와 조롱박세포 간 신경연접의 3차원 재구성을 시도하였다. 3차원 재구성에 앞서 염색방법과 절편 두께의 조절을 통해 고압전자현미경 하에서 신경연접의 적절한 관찰조건을 확립하고자 하였다. 관찰 결과, 절편의 두께가 증가하면 신경연접의 막, 소포와 같은 미세구조들의 겹침 현상이 나타나기 때문에 용이한 3차원 재구성을 위해서는 250 nm 두께의 절편을 제작하는 것이 적합한 것으로 판단되었다. 또한 절편제작 이전의 en bloc 염색 반응시간을 증가시키는 것이 절편제작 후 염색시간을 조절하는 것에 비해 contrast 증가에 더 효과적이었다. 이상의 결과로부터, 고압전자현미경을 이용하여 일련의 두꺼운 연속 절편을 촬영하고 3차원 재구성 프로그램을 이용하여 이미지들을 정렬하였으며 각각의 이미지에서 신경연접 막의 윤곽선을 그린 후 모든 윤곽선을 쌓아 올려 최종적으로 3차원 신경연접을 재구성하였다. 본 연구를 통하여 신경연접의 3차원 재구성에 있어 고압전자현미경의 적용 가능성을 검증하였고 관찰 조건을 확립하였다. 또한 고압전자현미경을 이용한 신경연접의 재구성은 많은 수의 연속절편 제작이 요구되는 기존의 방법에 비해 효율적이며 신경연접 연결형태에 관한 대규모의 정량 분석에 유용할 것으로 생각된다. 본 연구가 향후 고압전자현미경을 이용한 신경연접의 가소성 연구에 유용한 방법적 정보를 제공하기를 기대한다.

Telluric Acid가 흰쥐 간조직의 미세구조에 미치는 영향 (Ultrastructural Changes Induced by Telluric Acid in the Rat Liver)

  • 손석주;정영길;조승묵;백태경;최창도;최월봉
    • Applied Microscopy
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    • 제25권4호
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    • pp.83-103
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    • 1995
  • This experiment was carried out to investigate the effects of telluric acid on the histological and fine structural changes in the rat liver. Fischer 344 rats($150{\sim}200gm$) were used in this study as control and experimental groups. Telluric acid(5 mg/100 gm of body weight) suspensed in olive oil was given intraperitoneally to the animals of the experimental group and only olive oil to those of the control group. At the intervals of 3, 6 and 12 hours, 1, 2, 3, 5, 10, 20, 30 and 60 days after administration, the animals were sacrificed, and livers were obtained from the rats. For light microscopic examination of the liver, sections($5{\mu}m$) were stained with hematoxylineosin(H-E). For electron microscopic examination of the liver, sections were stained with uranyl acetate and lead citrate, finally examined with Zeiss EM 109 electron microscopes. The results obtained were as follows. 1. In the control group, round nucleus. well developed mitochondria, Golgi apparatus, rough endoplasmic reticulum(RER) and numerous glycogen particles were observed in the cytoplasm of the hepatocyte. In the cytoplasmic membranes of the hepatocyte, sinusoidal surface had numerous microvilli and cellular surface is combinated adjacent hepatocyte with desmosomes. The RER cisterns were dilated and zymogen granules were fewer than those of the dark cells. Kupffer cells with irregular nuclear membrane were observed. Fat storing cell and collagenous fiber bundle were observed in the Disse space. 2. Kupffer cell, inflammatory cells in the connective tissue of hepatic triad and lysosome were increased in the 3, 6, and 12 hour experimental group comparing with that of the control group. 3. In the 1 day experimental group, infiltration of inflammatory cells in interlobular connective tissue, dilatation of sinusoidal capillary and increasing of Kupffer cell were observed. Atropic change of hepatocyte and aggregation of glycogen particles in the cytoplasm of hepatocyte were observed. In this group, desmosome near bile canaliculi and collagenous fiber bundle in the Disse space were increased comparing with that of the 12 hours experimental group. In the 2 days experimental group, desmosome, lysosome, peroxisome and collagenous fiber bundle were increased comparing with that of the 1 day experimental group. Furthermore, lamellated bodies were also seen in the cytoplasm of the hepatocyte. 4. In 3 and 5 days experimental groups, transformations of hepatic cell cord and degeneration of the hepatocyte were markedly inclosed comparing with the all experimental groups. And damaged RER and mitochondria. collagenous fiber bundle were also inclosed comparing with that of the 2 days experimental group. Autophagosome and fat storing cells with large lipid droplets were also observed comparing with that of the 2 days experimental group. Tight junction and desmosome between the hepatocytes were separated. These degenerating changes were severe through the all experimental groups. 5. In the 10 and 20 days experimental groups, arrangement of hepatic cell cords and cell organelles of hepatocytes were similar to those of the control group. However, aggregation of glycogen particles, dilatation of sinusoidal capillary and infiltration of inflammatory cells remained. 6. In the 30 days experimental group, the tissue findings were similar to those of the control grout. But lamellated bodies in some hepatocytes and lysosome were remained in the cytoplasms of the Kupffer cells. In the 60 days experimental group, these all changes were recovered as the control group. In conclusion, telluric acid would directly induce the degenerative and necrotic changes on the hepatic tissue. However, these changes were perfectly recoverd in the 60 days experimental group as the control group.

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