• 제목/요약/키워드: immunoelectron microscopy

검색결과 29건 처리시간 0.021초

Immunoelectron Microscopic Localization and Analysis of Herpes simplex Virus Type 1 Antigens

  • Chung, Charles C.;Lee, Hyung-Hoan;Cho, Myung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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    • pp.714-720
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    • 2000
  • Antigens of Herpes simplex virus type 1 (HSV-1) strain F were immunoblotted to identify the most immunodominant one, and the localization of this antigen was then studied using immunoelectron microscopy. The 67.8 kDa antigen appeared to be the most immunodominant one in a mouse model, and it showed randomly scattered and partially clustered distribution on the surface of the virion. The localization study was performed using immunogold with polyclonal anti-HSV-1 sera produced from BALB/c mice, and immunofluorescence demonstrated that the viral products in the HSV-2 infected Vero cells were distributed throughout the infected host cell, however, mainly on the surface of the host membrane.

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한국재래산양 췌장내분비세포의 면역전자현미경적 연구 (Immunoelectron Microscopic Study on the Endocrine Pancreas of the Native Korean Goat)

  • 이재현;교본선춘;견태관;삼촌성;이형식
    • Applied Microscopy
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    • 제26권1호
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    • pp.67-77
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    • 1996
  • 한국재래산양의 췌장내분비세포에 대해 몇종의 항혈청을 이용하여 면역전자현미경적으로 관찰하여 다음과 같은 결론을 얻었다. 췌도에서는 glucagon (A), insulin (B), somatostatin (D) 및 pancreatic polypeptides (PP-I과 PP-II) 등 5종의 세포를 동정하였다. 이 중 A, B 및 D세포의 정태학적 특징은 다른포유동물의 그것과 유사하였고 D세포는 serotonin 면역성이 인정되었다. PP세포는 과립의 형태학적 특징으로 보아 두가지 형태가 인정되었으며, 제I형은 원형인 동질성의 과립 ($220{\sim}440nm$)을 가지며, 과립내용물과 한계막 사이에는 얇은 halo를 보였으나 제II형은 과립이 다형태성을 보이며 ($240{\sim}440nm$ major axis, $150{\sim}200nm$ minor axis) 과립내용물과 한계막 사이는 거의 밀착되어 있었다. 이상의 결과로 한국재래산양의 췌도에는 A, B, D, PP-I 및 PP-II 등 5형의 세포로 구성되어 있으며, 이 중 PP-I세포는 다른 포유류의 PP세포에, PP-II세포는 enterochromaffin cell에 상당할 것으로 생각되었다.

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계배 골격근 근원섬유형성에 관한 면역전자현미경적 연구 (Immunogold Electron Microscopy on Skeletal Myofibrilogenesis of Chick Embryo)

  • 하재정;김정옥
    • 한국동물학회지
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    • 제34권1호
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    • pp.31-43
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    • 1991
  • In the present study, the localization of myosin and the pathway of myofibrilogenesis of skeletal muscle cells in culture were investigated by immunocytochemical methods including immunoelectron microscopy and hybridoma formation. There were manly free ribosomes in the cytosol of the myoblast. At 48 hr of the culture, the myofilaments started to form and at 72 hr began to assemble to form myoabrils. At this time the Z band appeared, but the sarcomeres did not link together. Aker 96 hr of culture, adjacent sarcomeres linked together and shotved the typical striation of the skeletal muscle fiber. Myosin was synthesized in free ribosomes at 24 hr of culture and localized at many myofilaments at 48 hr and assembled mvoabrils at 72 hr and at 120 hr of the culture, myosin was localized at thick filaments of the A band.

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Monoclonal Antibody와 Immunogold 표지법에 의한 Cryptosporidium의 Sporozoites와 Merozoites의 공통항원의 구조적 위치 결정 (Ultrastructural Localization of a Common Antigen of Sporozoites and Merozoites of Cryptosporidium by Immunogold Labeling Technique Using a Monoclonal Antibody)

  • Cho, Myung-Hwan
    • 한국미생물·생명공학회지
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    • 제17권5호
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    • pp.499-503
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    • 1989
  • Cryptosporidium이 인체 내에서 복잡한 life cycle를 거치면서 각 stage마다 만들어지는 단백질 항원의 관계성에 관한 보고가 없어 Cryptosporldiosis에 대한 치료제 개발에 어려움이 있었다. 본 연구는 단일군 항체와 Immunogold labeling 기술을 이용하여 주요 extracellular stages인 sporozoites와 merozoites의 antigenic relatedness를 살펴보았다. BALB/C 쥐로부터 merozoites에 대한 단일군 항체(Jo3)를 분리하였으며 IgG3형이었다. 정제된 sporozoites를 SDS-PAGE로 분리한 후 Western blot을 이용하여 Jo3를 반응시킨 결과 3,500Daltons 크기의 sporozoites 항원을 인식하였다. Jo3를 Cryptosporidium에 감염된 tissue section에 반응시킨 후 Immunoelectron microscopy를 이용하여 3.5-kDa 항원의 위치와 sporozoites와 merozoites가 똑같은 단백질 항원을 만드는가를 추적해 본 결과 3.5-kDa 단백질 항원이 두 stages에서 공동으로 합성되는 것으로 나타났으며 이 항원은 표면과 세포질 내에 위치하고 있었다.

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흰쥐 눈물샘의 Prolactin 존재에 관한 면역전자현미경적 연구 (Immunoelectron Microscopic Localization of Prolactin in Rat Exolacrimal gland)

  • 박경호
    • Applied Microscopy
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    • 제23권1호
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    • pp.25-34
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    • 1993
  • Prolactin has been reported to be present in the tear film of humans and prolactin-like immunoreactivity has been detected by immunofluorescence in acinar cells of the lacrimal glands of humans and rats. The present study was aimed at clarifying the intracellular distribution of the prolactin-like immunoreactivity, using the electron microscope immunogold technique. The lacrimal gland acinar cells have two types of secretory granules: 1) Secretory granules containing flocculent materials irregularly shaped and are often coalesced. 2) Secretory granules are fairly round and contain homogenous materials of a moderate electron density. The density of the granular content varies even within a single cell. We found prolactin-like reactivity in secretory granules, some smaller cytosolic vesicles, Golgi cisternae and nuclei in acinar cells from intact glands of rat. Our present results are consistent with the conclusion that prolactin is present in lacrimal cells. The presence of prolactin reactivity in the nucleus suggests that prolactin may be a regulatory factor modulating gene expression.

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랫드 궁둥신경에서 말이집틈새의 형성과 Neurofascin 발현 연관에 대한 면역세포화학적연구 (Relationship between the formation of Schmidt-Lantermann incisure and the localization of neurofascin in rat sciatic nerve)

  • 최혜영;조익현;이종환;남상섭;장병준
    • 대한수의학회지
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    • 제49권4호
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    • pp.273-278
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    • 2009
  • Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the formation of Schmidt-Lantermann incisure in the sciatic nerve, the localization of neurofascin was studied with electron microscopy, immuno-fluorescence and immuno-electron microscopy. In the electron microscopy, the first formation of Schmidt-Lantermann incisure was checked at postnatal day 6 and the complete form of incisures traversing the whole myelin sheath began to be observed at postnatal day 8. In the immunofluorescence, neurofascin immunoreactive Schmidt-Lantermann incisures were first checked at postnatal day 6 and dramatically increased with aging by postnatal day 56. In the immunoelectron microscopy, neurofascin immunoreactive gold particles at the incisure forming sites were first observed at postnatal day 6 and the number of gold particles was increased as the animal was getting old by postnatal day 56. According to the present study, neurofascin is likely to have some relationships with Schmidt-Lantermann incisure formation.

계배 배양 출근세포에 있어서의 근원섬유 형성과정 :면역 전자현미경적 연구 (Myofibrillogenesis in Embryonic Chick Pectoral Muscle Cultures: An Immunoelectron Microscopic Study)

  • 하재청;김한도김동수
    • 한국동물학회지
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    • 제35권2호
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    • pp.149-160
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    • 1992
  • Using the protein A-gold complex, the mvoabrillogenesis and actin localization of cultured myoblast were invastisated. In the superstructural changes of mvogenic cell during differentiation, pectoral myoblasts contained large nucleus and numerous ribosomes but no myofibrils during the first 24 hr of cultures. Mvoblast initiated to differentiate at 3-day of culture contained the primitive myofibrillar structure. At 96 hr of culture, the mvofibrillar structure showed reletively discernable Z band but pools defined A, H and M bands. The feature of sarcomeric structure showed more defined form at cultur 5 day. In the aspect of actin localization, actin wvas diffusely detected throughout the cytoplasm of myogenic cell and nucleus during the proliferating stage. At 72 hr of culture, with the appearantc oi primitive mvofibrils, gold particles were observed in surrounding of myofibrils but still presented in overall of cytoplasm, especially in the surface and lumen of endoplasmic reticulum. With the gradual increase of culture time, local distribution of actin was readily detected within cytoplasm. In the 5-day specimen of cultures, gold particles precisely indicate the sites of actin localifation within the sarcomere. These results indicate the time of onset of myofibrill appearance and the biosynthetic and incorporation pathway of actin molecules into sarcomeric structure during myofibrillogenesis. Thus, in the present study, the first mvoabrillar structure was detected at culture 3 day, and the initiation of assembly into a typical sarcmeric structure was observed at culture 5 day. It seems, however, that the course of events on myofibrillogenesis of cultured myoblasts can be changed with great dependence of culture conditions including the number and groluth rate of mononucleated mvoblasts after seeding although the fundamental process shows identical appearances.

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세포 비병원성 소 설사병 바이러스의 이화학적 성상 조사 (Biophysical characteristics of a noncytopathic bovine viral diarrhea virus)

  • 권창희;Castro E Anthony;우희종
    • 대한수의학회지
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    • 제32권1호
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    • pp.77-82
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    • 1992
  • 세포 비병원성 소 설사병 바이러스가 감염된 우태아 폐세포를 이용 바이러스를 순수분리하여 이화학적 성상을 검사하였던 바 다음의 결과를 얻었다. 바이러스의 비중은 $1.090{\sim}1,114gm/cm^3$로 검출되었으며 $1,098gm/cm^3$에서 최대 감염가를 나타내었다. 면역 전자현미경을 이용한 형태학적 분석결과 30~80nm의 바이러스성 입자를 관찰할 수 있었으며 유전인자의 추출 분석결과 병원성 바이러스와의 전기영동상 유전자 규모에 있어서 차이는 인정되지 않았다. 그러나 비세포 병원성 소 설사병 바이러스는 병원성 바이러스에 비하여 세포외 배출이 적었으며 세포내에서 보다 많이 존재하는 것으로 관찰되었다.

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Influence of calcium ion on host cell invasion and intracellular replication by Toxoplasma gondii

  • Song, Hyun-Ouk;Ahn, Myoung-Hee;Ryu, Jae-Sook;Min, Duk-Young;Joo, Kyoung-Hwan;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • 제42권4호
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    • pp.185-193
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    • 2004
  • Toxoplasma gondii is an obligate intracellular protozoan parasite, which invades a wide range of hosts including humans. The exact mechanisms involved in its invasion are not fully understood. This study focused on the roles of $Ca^{2+}$ in host cell invasion and in T. gondii replication. We examined the invasion and replication of T. gondii pretreated with several calcium modulators, the conoid extrusion of tachyzoites. Calmodulin localization in T. gondii were observed using the immunogold method, and $Ca^{2+}$ levels in tachyzoites by confocal microscopy. In light microscopic observation, tachyzoites co-treated with A23187 and EGTA showed that host cell invasion and intracellular replication were decreased. The invasion of tachyzoites was slightly inhibited by the $Ca^{2+}$ channel blockers, bepridil and verapamil, and by the calmodulin antagonist, calmidazolium. We observed that calcium saline containing A23187 induced the extrusion of tachyzoite conoid. By immunoelectron microscopy, gold particles bound to anti-calmodulin or anti-actin mAb, were found to be localized on the anterior portion of tachyzoites. Remarkably reduced intracellular $Ca^{2+}$ was observed in tachyzoites treated with BAPTA/AM by confocal microscopy. These results suggest that host cell invasion and the intracellular replication of T. gondii tachyzoites are inhibited by the calcium ionophore, A23187, and by the extracellular calcium chelator, EGTA.

완두 자엽에서 소포체 말단의 팽창에 의한 단백과립 발달 (Terminal Dilation and Transformation of the Protein-filled ER to Form Protein Bodies in Pea (Pisum sativum L. var, exzellenz) Cotyledons)

  • 정병갑
    • Applied Microscopy
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    • 제29권4호
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    • pp.499-509
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    • 1999
  • 완두 종자에 축적되는 저장물질은 주로 전분과 단백질로서 이러한 저장물질 때문에 고정이나 전자현미경 관찰시료를 제작하기가 쉽지 않다. 따라서 자엽을 얇게 절편을 만들고 효소를 사용하여 단일세포로 분리한 다음 고정하여 관찰하였다. 완두의 저장단백질이 축적되는 단백질 저장 액포는 종자발달의 이른 시기에 기존의 액포를 둘러싸고 발달하게 되므로서 액포는 수축되고 단백질 저장 액포는 점점 발달하여 그 가장자리에 단백질 덩어리가 축적되게 된다. 이와는 별도로 종자발달의 이른 시기에 조면소포체의 내강에 전자밀도가 높은 단백질이 축적되기 시작하여 늦은 시기에 이 소포체의 끝이 부풀어서 구형의 단백과립으로 발달하였다. 완두종자의 저장단백질은 주로 vicilin과 legumin으로서 단백과립에 대한 면역세포화학적 방법으로 확인한 결과 vicilin은 세포질에 발달된 작은 단백과립과 단백질 저장액포의 가장자리에 축적된 단백질 덩어리에 모두 반응하였으나 legumin은 세포질의 단백과립에만 반응하였다. 또한 소포체에 존재하는 단백질인 Bip은 단백질 저장액포에 축적된 단백질 덩어리의 안쪽 가장자리에만 반응하였다. 이는 단백질이 활발하게 축적되고있는 시기에 특징적으로 작용하는 Bip의 기능과 관련되는 것으로 사료된다.

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