• 제목/요약/키워드: Lead acetate

검색결과 216건 처리시간 0.028초

방사선이 흰쥐 갑상샘 소포곁세포에 미치는 영향에 대한 미세구조적 연구 (Electron Microscopic Study on the Parafollicular Cells of the Thyroid Gland of the Head and Neck-Irradiated Rats)

  • 김용식;양남길;안의태;고정식;박경호;김진국
    • Applied Microscopy
    • /
    • 제22권2호
    • /
    • pp.1-14
    • /
    • 1992
  • This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

  • PDF

이질아메바(Entamoeba histolytica)의 미세구조 및 효소활성에 관한 전자현미경적 연구 (Electron-microscopic studies on fine structure and enzyme activity in the axenic and conventional strains of Entamoeba histolytica)

  • 용태순;정평림;이근태
    • Parasites, Hosts and Diseases
    • /
    • 제23권2호
    • /
    • pp.269-284
    • /
    • 1985
  • The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.

  • PDF

Methylene Blue의 방사선방어결과 2. Methylene Blue가 $\gamma$선에 조사한 흰줘의 간 및 심장조직에 미치는 전자현미경적 연구 (Radioprotective Effect of Methylene Blue: 2. Electron Microscopy of the Effect of Methylene Blue on the Liver and Heart of Rats following Gamma-Irradiation)

  • Sang Yul Nam;Seung Han Chang
    • 한국동물학회지
    • /
    • 제12권4호
    • /
    • pp.114-122
    • /
    • 1969
  • 成體 albino 흰쥐를 實驗群과 對照群으로 나누어 對照群에는 0.9% 生理食鹽水를, 實驗群에는 methylene blue(38mg/kg, pH 7.4)를 各各 腹腔에 注射한 後 約 30 分後에 總線量 360 rads의 $^{60}Co-\gamma$ 線을 一時全身照射하였다. 照射後 64時間區 와 212時間區로 나누어서 肝 및 心臟組織을 觀察하였다. 照射直後 肝 및 心臟組織을 6% glutaraldehyde (pH 7.4)와 1% osmium tetroxide (pH 7.4)로 冷室에서 二重固定하였으며 脫水한 後 Epon 812로 胞埋하여 MT-2 Porter Blum ultramicrotome 으로 超薄切斷하여 切片을 만들었다. 이를 uranyl acetate 와 lead citrate 로 二重染色한후 Hi9tachi HU-11 E型 電子顯微鏡으로 觀察하였다. 64時間區의 methylene blue 處理群과 對照群에서 肝組織은 顯著한 組織學的 變化의 差를 나타냈다. 다시 말하면 methylene blue 處理群에 比하여 對照群에서는 mitochondria 의 膨大, cristae 의 切斷, endoplasmic reticulum 의 破壞를 觀察할 수 있었으며 또한 endoplasmic reticulum 에 glycogen 粒子의 蓄積을 觀察할수 있었다. 한편 methylene blue 處理群에 比하여 對照群의 212時間區에서도 같은 變化樣相을 나타냈으나 endoplasmic reticulum 에 많은 vacuole 이 形成되었음을 觀察할 수가 있었다. 그러나 methylene blue 處理群은 正當群($\gamma$ 線의 照射와 methylene blue를 處理하지 않음)에 比하여 若干의 差가 觀察되었을 뿐 顯著한 組織學的變化는 나타나지 않았다. 心臟組織에서는 兩群 卽 實驗群과 正當群사이에 顯著한 差는 別로 觀察할 수 없었으나 methylene blue 群에 比하여 對照群에 若干의 變化가 觀察되었다. 다시말하면 methylene blue 處理群에 比하여 64時間區의 對照群에서는 mitochondria의 若干의 膨大, cristae 에 若干의 切斷을 觀察할수 있었고 212時間區의 對照群에서는 sarcoplasmic reticulum에서 若干의 液胞와 glycogen 粒子의 若干의 增加를 觀察할수 있었다. 이로 미루어 보아 methylene blue 가 放射線에 對하여 防禦 果가 있는 것으로 思料된다.

  • PDF

X-선 조사를 받은 흰쥐 망막의 색소상피세포와 기저복합층의 미세구조 (Fine Structure on the Pigment Epithelial Cell and the Bruch's Membrane of the Rat Retina after X-Irradiation)

  • 고정식;신기호;안의태;양남길;박경호;김진국
    • Applied Microscopy
    • /
    • 제23권2호
    • /
    • pp.11-26
    • /
    • 1993
  • This experiment was performed to study the morphological responses of the pigment epithelial cell and the Bruch's membrane of the retina of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80cm, and the. dose rate was 200 rads/min. The experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Under anesthesia, 1% glutaraldehyde-1% paraformaldehyde solution(0.1M Millonig's phosphate buffer, pH 7.3) was perfused through the left ventricle and ascending aorta. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.3) and 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH7.3), and embedded in araldite mixture. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. The morphological changes of the pigment epithelial cells were not pronounced after exposure to 3,000 rads of X-ray. But on the 6th hour after exposure to 6,000 rads of X-ray, bulging nuclear membrane protruding into the cytoplasm and nuclear chromatin clumped into numerous masses along the nuclear membrane were observed. At the 2nd and 6th day post-irradiation, partial cytolysis or necrosis were seen. 2. The thickness of the Bruch's membrane of the experimental groups were increased in the time and dose range covered by this study, and splitting or diffusing basal laminae of the choriocapillary layer were observed frequently in the experimental group. Above results suggest that large amount(6,000 rads) of head irradiation induce direct hazardous effects on the pigment epitherial cells and Bruch's membrane of the retina of the rat, but pigment epithelial cells are more radioresistant than Bruch's membrane.

  • PDF

Neuroprotective effects of antioxidant constituents isolated from Opuntia ficus-indica var. saboten Makino

  • K.J. Jung;Lee, E.H.;Kim, H.J.;Lee, J.Y.;Y.S. Song;Lee, Y.H.;J. Cho;Park, M.;Park, H.
    • 한국응용약물학회:학술대회논문집
    • /
    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
    • /
    • pp.63-63
    • /
    • 2003
  • Opuntia ficus-indicavar. saboten Makino (Cactaceae) is a tropical or subtropical plant that has been widely used as folk medicine for the treatment of diabetes, asthma, burn, edema and gastritis. The purposes of the present study were to identify antioxidant constituents from fruits and stems of the plant cultivated in Cheju island, Korea, and examine their in vitro neuroprotective activities. Using a chromatographic fractionation method, ten chemical constituents were isolated from ethyl acetate extracts. By means of chemical and spectroscopic methods, those were identified as eight flavonoids such as kaempherol (a), quercetin (b), kaempferol 3-methyl ether (c), quercetin 3-methyl ether (d), narcissin (e), dihydrokaernpferol (f), dihydroquercetin (g) and erioclictyol (h), and two terpenoids such as 3-oxo-${\alpha}$-ionol-${\beta}$-d-glucopyranoside (i) and roseoside (j). Among the isolated compounds, comrounds c~e and h~j were those reported for the first titre from the plant. Compounds b, d and g showed DPPH free radical scavenging activities with IC$\sub$50/ values of 28, 19 and 31, ${\mu}$M respectively. Compounds d and g also inhibited iron-dependent lipid peroxidation with IC$\sub$50/ values of 2.4 and 3.5 ${\mu}$M. In a primary rat cortical neuronal cell culture system, compounds b, d and g inhibited xanthine/xanthine oxidase-induced (IC$\sub$50/ values of 18.2, 2.1 and 54.6 ${\mu}$M) and H$_2$O$_2$-induced (IC$\sub$50/ values of 13.6, 1.9 and 25.7 ${\mu}$M) cytotoxicities. In addition, compounds d and g inhibited NMDA-induced excitotoxicity by 21 and 33%, and only compound d inhibited growth factor withdrawal-induced apoptosis by 31% at a tested concentration of 3 ${\mu}$M. The results suggest that the antioxidant constituents with in vitroneuroprotective activities may serve as lead chemicals for the development of neuroprotective agent.

  • PDF

수산식품 중 말라카이트그린 및 류코말라카이트그린의 분석 (Analysis of malachite green and leuco-malachite green in sea food)

  • 최동미;홍순근;임무혁;정지윤;장문익;박건상;홍무기;우건조
    • 분석과학
    • /
    • 제19권2호
    • /
    • pp.142-148
    • /
    • 2006
  • 고속액체크로마토그라피를 이용하여 어류 중 말라카이트그린 및 류코말라카이트그린의 최적 분석조건을 확립하고 국내유통 수산식품을 분석하였다. 균질화한 시료를 아세토니트릴로 추출하고 디클로로메탄에 분산시켜 농축 후 메탄올에 용해하여 분석하였다. 이동상으로는 아세토니트릴과 아세테이트 완충용액의 혼합용액을 사용하였으며, 검출파장은 620 nm이었다. 류코말라카이트그린을 말라카이트그린과 동시에 분석하기 위하여 산화납(IV) 컬럼을 분석컬럼 뒤에 연결하여 류코말라카이트그린을 말라카이트그린으로 산화하였다. 상관계수($r^2$)는 말라카이트그린이 0.9989, 류코말라카이트그린이 0.9995이었으며, 검출한계는 신호대 잡음비 3 이상에서 말라카이트그린과 류코말라카이트그린의 합으로서 0.005 mg/kg이었다. 회수율은 84~98%(CV는 3~16%)로 만족할 만한 수준이었다. 분석결과 잉어 및 붕어 서료에서 류코말라카이트그린이 검출되었다.

Zinc 대사와 관련된 Paneth 세포활성의 변화에 관한 조직화학적 연구 (Do Paneth Cells Regulate the Zinc Body Burden?)

  • 조승묵;김성준;박승국;강태천;원무호
    • Applied Microscopy
    • /
    • 제30권4호
    • /
    • pp.357-365
    • /
    • 2000
  • 포유류 장생 상피세포의 하나인 파네스세포(Paneth cell)는 여러 가지 금속류를 함유하고 있으며, 생체가 과잉의 이들 금속류에 노출되거나 부족할 때에 장 내강을 통해 제거하거나 흡수함으로써 금속류의 항상성에 기여한다고 알려져 있다. 이번 연구는 이러한 연구보고에 근거하여 금속류중에서 zinc를 실험적으로 과량투여한 후 파네스세포내 zinc의 분포에 어떤 변화가 초래되는 지를 광학 및 전자현미경적 autometallography(AMG)로 관찰함으로써 파네스세포의 zinc와 관련된 세포생물학적 기능을 규명하고자 하였다. Wistar 랫드에 체중 당 20mg의 zinc chloride를 생리식염수 5ml에 녹여 복강주사한 후 2시간에 이르러 0.1 M phosphate buffer(PB)에 녹인 0.5% sodium sulphide-3% glutaraldehyde 흔합액으로 관류고정하였다. 회장(ileum)의 일부를 Dry Ice나 $CO_2$ gas로 얼린 후 cryostat을 이용하여 $20{\mu}m$ 두께의 조직절편을 만들어 및 단계의 AMG법을 시행하였다. 이와는 별도로 EM용으로 선택된 회장절편은 일련의 전자현미경 표본제작과정을 거쳐, 100nm두께의 thin section을 만들어 uranyl-lead 이중염 색 추 전자현미경으로 관찰하였다. Zinc를 투여한 동물에서 관찰된 소견은 생리식염수만을 투여한 대조군의 것과 비교할 때 큰 차이를 보였다. 우선 대조군에서는 파네스세포의 꼭대기(apex)부위에서 낱알모양을 띤 AMG양성반응 구조물(AMG grain)들이 분비과립과 사이토졸(cytosol)에서 관찰되었고, 세포사이공간에서도 적은 양의 grain이 분포하고 있었다. 반면 zinc를 투여한 랫드의 파네스세포에서는 AMG 입자가 이상의 부위에서 훨씬 많은 양의 grain이 분포해 있었으며, 특히 분비과립에서는 가장 높은 농도로 관찰되었다. 더욱이 대조군에서 관찰되지 많았던 장샘의 분비관(Lieberkuhn crypt)및 고유판 혈관의 내에서도 많은 grain이 관찰되었다. 이상의 결과를 요약하면 과량의 zinc에 노출된 생체는 혈관을 경유하여, 파네스세포의 분비과립을 매개로 하여, 장관의 내강쪽으로 과잉의 zinc 를 피내고 있는 모습을 보인다. 따라서 연구자는 파네스세포가 생체내 zinc의 양을 조절하는데 중요한 역할이 있을 것으로 본다.

  • PDF

교모세포종 세포주 U-87에서 세포내 PKC 농도와 종양침습성과의 상관 관계 (The Relationship between Intracellular Protein Kinase C Concentration and Invasiveness in U-87 Malignant Glioma Cells)

  • 지철;조경근;이경진;박성찬;조정기;강준기;최창락
    • Journal of Korean Neurosurgical Society
    • /
    • 제30권3호
    • /
    • pp.263-271
    • /
    • 2001
  • 교모세포종은 비교적 흔한 원발성 뇌종양이며 생물학적 특성상 빠른 성장률을 보이는 것 외에 침습성이 강하여 종양과 인접한 부분을 파괴 시킬 뿐 아니라 직접접촉하지 않는 부분의 파괴도 일어나게 되어 그 결과 치료 예후가 매우 불량한 것으로 되어 있다. 이러한 불량한 예후를 개선 시키기 위해서는 이들 종양의 침습에 대한 기전의 정확한 이해가 필요하며 이를 이용한 새로운 치료방법이 요구된다할 것이다. Protein kinase C(PKC)는 세포내 신호전달체제 과정에서 매우 중요한 역할을 하는 효소로 세포막 수용체 신호를 핵으로 전달하는 역할을 하며 세포내 여러 생물학적 작용이 알려져 있다. 본 실험은 종양침습과 연관하여 세포내 PKC가 어떠한 작용을 하는지에 대해서 악성교종 세포를 대상으로 하여 알아보고자 하였다. 따라서 PKC가 종양침습에 중요한 역할을 할 것이라는 가설을 세웠고 이 가설을 증명하기 위해 세포내 PKC농도를 길항제 및 촉진제를 이용하며 높고 낮게 조절함으로써 그에 따른 침습성의 변화를 살펴보았다. 방법으로는 교모세포종 세포주인 U-87 세포를 약제로 처리한 후 인위적으로 조절된 세포내의 PKC에 대해 효소의 활성도를 측정하였고 침습성은 matrigel artificial basement membrane assay 및 tumor spheroid fetal rat brain aggregate(FRBA) confrontation assay를 이용하여 측정하였다. 결과로 PKC의 길항제인 tamoxifen과 hypericin으로 처치한 세포는 PKC의 활성과 침습도가 모두 감소하였으며 이는 약제농도에 비례하여 나타났다. 반면 PKC 자극제인 TPA로 처치된 세포는 증가된 PKC 활성도나 침습도을 보이지 않았다. 이러한 결과를 종합해 보았을 때 PKC는 종양세포의 침습성에 중요한 역할을 함을 알 수 있었으며 PKC의 길항제는 종양 치료에 유용한 화학 요법 제가 될 수 있을 것으로 사료된다.

  • PDF

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
    • /
    • 제25권4호
    • /
    • pp.83-103
    • /
    • 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.

  • PDF

총담관결찰후 집토끼 Paneth세포의 변화에 대한 면역전자현미경적 연구 (Immunoelectron Microscopic Study on the Paneth Cell of Rabbit after the Common Bile Duct Ligation)

  • 박경호;조휘동;양남길;안의태;고정식;김진국
    • Applied Microscopy
    • /
    • 제24권2호
    • /
    • pp.78-92
    • /
    • 1994
  • Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

  • PDF