• The Korean Journal of Physiology and Pharmacology
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• 제8권2호
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• pp.83-88
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• 2004

The purpose of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Large unilamellar vesicles (OPGTL) were prepared with total lipids extracted from cultured Porphyromonas gingivalis outer membranes (OPG). The anthroyloxy probes were located at a graded series of depths inside a membrane, depending on its substitution position (n) in the aliphatic chain. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine effects of chlorhexidine digluconate on differential rotational mobility, while changing the probes' substitution position (n) in the membrane phospholipids aliphatic chain. Magnitude of the rotational mobility of the intact six membrane components differed depending on the substitution position in the descending order of 16-(9-anthroyloxy)palmitic acid (16-AP), 12, 9, 6, 3 and 2-(9-anthroyloxy)stearic acid (12-AS, 9-AS, 6-AS, 3-AS and 2-AS). Chlorhexidine digluconate increased in a dose-dependent manner the rate of rotational mobility of hydrocarbon interior of the OPGTL prepared with total lipids extracted from cultured OPG, but decreased the mobility of membrane interface of the OPGTL. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

• Applied Microscopy
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• 제37권2호
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• pp.65-72
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• 2007

참붕어의 난자형성과정과 수정란 난막의 미세구조를 광학현미경과 전자현미경으로 관찰한 결과는 다음과 같다. 참붕어에서 난원세포의 세포질은 호염기성이었고 핵막 내에 많은 인들이 분포하고 있었다. 제1난모세포의 경우 난황포가 단지 난세포 가장자리에만 배열되어 있었고 난막의 형성은 관찰되지 않았다. 제2난모세포에서는 난막이 형성되었고 제1난모세포에 비해서 난황포가 점점 핵쪽으로 증식된 경향을 보였다. 발생이 진행됨에 따라서 호염기성 물질들은 점점 감소하는 경향을 보였고 후에 난막주위에만 국한적으로 분포하였으며, 난막의 두께와 난자의 크기는 점점 증가되었다. 또한 성숙란으로 발생이 진행되면서 난황포들은 난황괴로 변화되었다. 참붕어의 수정란은 타원형으로 부착성이었으며 동물극쪽에서 난문이 관찰되었다. 수정란의 난막은 모두 3층으로, 부착성인 외층, 전자밀도가 서로 다른 6층상구조를 가진 중층 및 전자밀도가 높은 내층으로 구성되어 있었으며 난막표면은 부착성 섬유상 구조물로 덮여 있었다. 이상과 같이 참붕어의 난자형성과정은 생식세포의 크기 증가, 난황의 형성과 축적 및 세포질의 호염기성물질 감소로 될 수 있으며 수정란 난막의 미세구조적 특징들은 경골어류의 계통분류학에서 분류형질도 이용될 수 있을 것으로 생각된다.

• Applied Microscopy
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• 제39권3호
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• pp.237-243
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• 2009

담수산 잉어과(Cyprinidae), 황어아과(Leuciscinae)에 속하는 버들치(Chinese minnow; Rhynchocypris oxycephalus)의 난자형성과정과 성숙난 난막의 미세구조를 광학현미경과 전자현미경으로 관찰한 결과는 다음과 같다. 난원세포의 세포질은 호염기성이었고 핵 내에 다수의 인들이 분포하고 있었다. 제1난모세포의 경우 난황포가 단지 난세포 가장자리에만 배열되어 있었고 난막의 형성은 관찰되지 않았다. 제2난모세포에서는 난막이 형성되었고 제1난모세포에 비해서 난황포가 점점 핵 쪽으로 증식된 경향을 보였다. 발생이 진행됨에 따라서 호염기성 물질들은 점점 감소하는 경향을 보였고 후에 난막 주위에만 국한적으로 분포하였으며, 난막의 두께와 난자의 크기는 점점 증가되었다. 배란된 성숙난 난막의 표면은 미세융모와 같은 구조물들에 의하여 덮혀 있었고 동물극 쪽에 깔떼기 형태의 난문이 발견되었다. 난막의 단면을 확인한 결과 부착성 외층과 섬유상 내층 모두 2층으로 구성되어 있었다. 이상과 같이 버들치의 난자형성과정은 난세포의 크기 증가, 난황낭의 축적, 염기성 물질의 감소, 난막발달 및 두께의 증가로 요약될 수 있으며 이 패턴은 잉어과의 공통적인 특징으로 생각된다. 그러나 배란된 성숙난 난막의 미세구조적 특징들은 이 종만이 가지는 종특이성으로 종을 분류하는 데 이용될 수 있다.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.2095-2105
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• 2018

In our previous studies, we have identified several in vivo-induced antigens and evaluated their potential as subunit vaccine candidates in a murine model, in which the recombinant protein GalT showed the most potent immunogenicity and immunoprotective efficacy against Actinobacillus pleuropneumoniae. To exploit a more efficient way of delivering GalT proteins, in this study, we employed the widely studied E. coli outer membrane vesicles (OMVs) as a platform to deliver GalT protein and performed the vaccine trial using the recombinant GalT-OMVs in the murine model. Results revealed that GalT-OMVs could elicit a highly-specific, IgG antibody titer that was comparable with the adjuvant GalT group. Significantly higher lymphocyte proliferation and cytokines secretion levels were observed in the GalT-OMVs group. 87.5% and 50% of mice were protected from a lethal dose challenge using A. pleuropneumoniae in active or passive immunization, respectively. Histopathologic and immunohistochemical analyses showed remarkably reduced pathological changes and infiltration of neutrophils in the lungs of mice immunized with GalT-OMVs after the challenge. Taken together, these findings confirm that OMVs can be used as a platform to deliver GalT protein and enhance its immunogenicity to induce both humoral and cellular immune responses in mice.

• Journal of Photoscience
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• 제12권2호
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• pp.67-73
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• 2005

Fluorescent probe techniques were used to evaluate the effect of dibucaine.HCl on the physical properties (transbilayer asymmetric lateral mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. An experimental procedure was used based on selective quenching of 1,3-di(l-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Dibucaine.HCl increased the bulk lateral mobility, and annular lipid fluidity in SPMV lipid bilayers, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. The magnitude of increasing effect on annular lipid fluidity in SPMV lipid bilayer induced by dibucaine.HCl was significantly far greater than magnitude of increasing effect of the drug on the lateral mobility of bulk SPMV lipid bilayer. It also caused membrane proteins to cluster. These effects of dibucaine.HCl on neuronal membranes may be responsible for some, though not all, of the local anesthetic actions of dibucaine.HCl.

• Molecules and Cells
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• 제19권3호
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• pp.356-364
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• 2005

Intramolecular excimer formation of 1,3-di(1-pyrenyl) propane(Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of lateral and rotational mobilities of bulk bilayer structures of synaptosomal plasma membrane vesicles (SPMVs) from the bovine cerebral cortex. Ethanol increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in the SPMVs. Selective quenching of both DPH and Py-3-Py by trinitrophenyl groups was used to examine the range of transbilayer asymmetric rotational mobility and the rate and range of transbilayer asymmetric lateral mobility of SPMVs. Ethanol increased the rotational and lateral mobility of the outer monolayer more than of the inner one. Thus ethanol has a selective fluidizing effect within the transbilayer domains of the SPMVs. Radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py was used to examine both the effect of ethanol on annular lipid fluidity and protein distribution in the SPMVs. Ethanol increased annular lipid fluidity and also caused membrane proteins to cluster. These effects on neuronal membranes may be responsible for some, though not all, of the general anesthetic actions of ethanol.

• The Korean Journal of Physiology and Pharmacology
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• 제16권4호
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• pp.255-264
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• 2012

The structures of the intact synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortexs, and the outer and the inner monolayer separately, were evaluated with 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1,3-di(1-pyrenyl)propane (Py-3-Py) as fluorescent reporters and trinitrophenyl groups as quenching agents. The methanol increased bulk rotational and lateral mobilities of SPMVs lipid bilayers. The methanol increased the rotational and lateral mobilities of the outer monolayers more than of the inner monolayers. n-(9-Anthroyloxy)stearic acid (n-AS) were used to evaluate the effect of the methanol on the rotational mobility at the 16, 12, 9, 6, and 2 position of aliphatic chains present in phospholipids of the SPMVs outer monolayers. The methanol decreased the anisotropy of the 16-(9-anthroyloxy)palmitic acid (16-AP), 12-(9-anthroyloxy)stearic acid (12-AS), 9-(9-anthroyloxy)stearic acid (9-AS), and 6-(9-anthroyloxy)stearic acid (6-AS) in the SPMVs outer monolayer but it increased the anisotropy of 2-(9-anthroyloxy)stearic acid (2-AS) in the monolayers. The magnitude of the increased rotational mobility by the methanol was in the order at the position of 16, 12, 9, and 6 of aliphatic chains in phospholipids of the outer monolayers. Furthermore, the methanol increased annular lipid fluidity and also caused membrane proteins to cluster. The important finding is that was far greater increase by methanol in annular lipid fluidity than increase in lateral and rotational mobilities by the methanol. Methanol alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that methanol, in additions to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membranes lipids.

• IMMUNE NETWORK
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• 제23권1호
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• pp.3.1-3.14
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• 2023

Microvilli are outer membrane organelles that contain cross-linked filamentous actin. Unlike well-characterized epithelial microvilli, T-cell microvilli are dynamic similar to those of filopodia, which grow and shrink intermittently via the alternate actin-assembly and -disassembly. T-cell microvilli are specialized for sensing Ags on the surface of Ag-presenting cells (APCs). Thus, these finger-shaped microprotrusions contain many signaling-related proteins and can serve as a signaling platforms that induce intracellular signals. However, they are not limited to sensing external information but can provide sites for parts of the cell-body to tear away from the cell. Cells are known to produce many types of extracellular vesicles (EVs), such as exosomes, microvesicles, and membrane particles. T cells also produce EVs, but little is known about under what conditions T cells generate EVs and which types of EVs are released. We discovered that T cells produce few exosomes but release large amounsts of microvilli-derived particles during physical interaction with APCs. Although much is unanswered as to why T cells use the same organelles to sense Ags or to produce EVs, these events can significantly affect T cell fate, including clonal expansion and death. Since TCRs are localized at microvilli tips, this membrane event also raises a new question regarding long-standing paradigm in T cell biology; i.e., surface TCR downmodulation following T cell activation. Since T-cell microvilli particles carry T-cell message to their cognate partner, these particles are termed T-cell immunological synaptosomes (TISs). We discuss the potential physiological role of TISs and their application to immunotherapies.

• 한국수산과학회지
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• 제33권5호
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• pp.403-407
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• 2000

징거미새우의 정소는 간췌장의 배면 측상부에서 심장사이에 위치하며, 한 쌍의 소엽으로 이루어져 있고 각 소엽의 선단은 서로 연결되어 있다. 그리고 각 소엽은 수많은 세정관이 결합조직으로 연결되어 있다. 각 세정관은 생식세포와 지주세포가 밀착되어 있는 생식세포대를 제외한 부분은 낮은 입방상피에 의해 내강이 형성되어 있고, 세정관의 제일 바깥은 단층편평상피로 둘러싸여 있다. 세정관 사이에는 결합조직 외에 Leydig cell-like cell도 관찰된다. 단층편평상피세포는 얇은 측면으로 연결되어 있고 종종 이웃한 세포와 포개져 있다. 지주세포는 생식세포에 비해 그 수가 현저히 적고, 대부분 생식세포대의 가장자리에 위치하고 있다. 지주세포의 세포질은 기저판과 접해 있어 기저면과 생식세포 사이는 지주세포의 원형질에 의해 분리되어 있다. 지주세포의 핵은 대부분 각져 있고, 크고 현저한 인이 핵의 중심부에 위치하고 있다. 낮은 입방상피세포는 세정관의 기저판과 접하고 있는 기저면과 일부는 생식세포대와 내강 사이에 위치하고 있다. 입방상피세포의 세포질에서는 횡 방향의 크리스테가 발달된 미토콘드리아, 층상구조의 조면소포체 그리고 Golgi 복합체들이 매우 발달되어 있다. 그리고 조면소포체와 Golgi 복합체의 볼록한 형성면 사이에는 전이소낭이, Golgi 복합체의 성숙면에서는 분비소낭이 관찰되며, 상피세포의 선단에 수많은 홈들이 존재하는 점으로 보아 exocytosis에 의해 내강으로의 물질 분비가 이루어지는 것으로 판단된다.

• The Korean Journal of Physiology and Pharmacology
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• 제16권6호
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• pp.413-422
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• 2012

The purpose of this study is to investigated the mechanism of pharmacological action of local anesthetic and provide the basic information about the development of new effective local anesthetics. Fluorescent probe techniques were used to evaluate the effect of lidocaine HCl on the physical properties (transbilayer asymmetric lateral and rotational mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex, and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. An experimental procedure was used based on selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) and 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Lidocaine HCl increased the bulk lateral and rotational mobility of neuronal and model membrane lipid bilayes, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. Lidocaine HCl increased annular lipid fluidity in SPMV lipid bilayers. It also caused membrane proteins to cluster. The most important finding of this study is that there is far greater increase in annular lipid fluidity than that in lateral and rotational mobilities by lidocaine HCl. Lidocaine HCl alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that lidocaine, in addition to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membrane lipid.