• Bulletin of the Korean Chemical Society
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• v.35 no.12
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• pp.3509-3513
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• 2014

Phospholipase D (PLD) catalyzed the generation of phosphatidic acid (PA) from phosphatidylcholine (PC) at the outer layer of the vesicles prepared through layer by layer via a double emulsion technique. The generation induced a curvature change in the vesicles, which eventually led them to fuse each other. The ratio of two-fatty-acid-tail ethanolamine (PE) to one-fatty-acid-tail ethanolamine (PE) was found to acquire the condition where the mixed-phospholipid vesicles were stable identically with pure two-fatty-acid-tail PC. The effect of the outer-layer mixture on the PLD-induced vesicle fusion was investigated using the fluorescence intensity change. 8-Aminonaph-thalene-1,3,6-trisulfonic acid disodium salt (ANTS) and p-Xylene-bis(N-pyridinium bromide) (DPX) were encapsulated in the vesicles, respectively, for the quantification of the fusion. The fluorescence scale was calibrated with the fluorescence of a 1/1 mixture of ANTS and DPX vesicles in NaCl buffer taken as 100% fluorescence (0% fusion) and the vesicles containing both ANTS and DPX as 0% fluorescence (100% fusion), considering the leakage into the medium studied directly in a separate experiment using vesicles containing both ANTS and DPX. The fusion data for each composition were acquired with the subtraction of the leakage from the quenching. From the monitoring, the vesicle fusion caused by the PLD reaction seems dominantly to occur rather than the vesicle lysis, because the composition effect on the fusion was observed identically with that on the change in the vesicle structure. Furthermore, the diameter measurements also support the fusion dominancy.

• Bulletin of the Korean Chemical Society
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• v.33 no.12
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• pp.3998-4002
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• 2012

With continuing progress of nanotechnologies and various applications of nanoparticles, one needs to develop a quick and fairly standard assessment tool to evaluate cytotoxicity of nanoparticles. However, much cytotoxicity studies on the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Here, we propose a simple screening method for the analysis of the interaction between several AgNPs (5.3 to 64 nm) and fluorescence-dye containing vesicles ($12{\mu}m$) acting as a biomimetic cell-membrane. Fluorescence-dye containing vesicle was prepared using a fluorescence probe (1,6-diphenyl-1,3,5-hexatryene), which was intercalated into the lipid bilayer due to their hydrophobicity. Zeta potential of all materials except for bare-AgNPs (+32.8 mV) was negative (-26 to -54 mV). The morphological change (i.e., rupture and fusion of vesicle, and release of dye) after mixing of the vesicle and AgNPs was observed by fluorescence microscopy, and fluorescence image were different with coating materials and surface charge of x-AgNPs. In the results, we found that the surface charge of nanoparticles is the key factor for vesicle rupture and fusion. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

• Molecules and Cells
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• v.43 no.6
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• pp.501-508
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• 2020

In eukaryotes, membraneous cellular compartmentation essentially requires vesicle trafficking for communications among distinct organelles. A donor organelle-generated vesicle releases its cargo into a target compartment by fusing two distinct vesicle and target membranes. Vesicle fusion, the final step of vesicle trafficking, is driven intrinsically by complex formation of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). Although SNAREs are well-conserved across eukaryotes, genomic studies revealed that plants have dramatically increased the number of SNARE genes than other eukaryotes. This increase is attributed to the sessile nature of plants, likely for more sensitive and harmonized responses to environmental stresses. In this review, we therefore try to summarize and discuss the current understanding of plant SNAREs function in responses to biotic and abiotic stresses.

• Korean Chemical Engineering Research
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• v.53 no.6
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• pp.672-676
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• 2015

Spherical phospholipid bilayers, vesicles, were formed with respect to phase of each layer via a double emulsion technique. The conversion of phosphatidylcholine (PC) to phosphatidic acid (PA) at the outer layer, caused by phospholipase D (PLD), induced a curvature change in the vesicles, which eventually led them to fuse each other. The effect of the lipid layer physical-properties on the PLD-induced vesicle fusion was investigated using the fluorescence intensity change. 8-Aminonaphthalene-1,3,6-trisulfonic acid disodium salt(ANTS) and p-Xylene-bis(N-pyridinium bromide)(DPX) were encapsulated in the vesicles, respectively, for the quantification of the fusion. The fluorescence scale was calibrated with the fluorescence of a 1/1 mixture of ANTS and DPX vesicles in NaCl buffer taken as 100% fluorescence (0% fusion) and the vesicles containing both ANTS and DPX as 0% fluorescence (100% fusion), considering the leakage into the medium studied directly in a separate experiment using vesicles containing both ANTS and DPX. It was observed that the fusion occurred to the liquid-phase of the inner layer only. The fusion behaviors were very similar for both solid and liquid of the outer layer. However, the leakage was faster for the solid-phase outer-layer than the liquid-phase outer-layer. The difference in the leakage seems to be caused by the lipid concentration and the lateral diffusivity in the layer.

• Molecules and Cells
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• v.43 no.4
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• pp.313-322
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• 2020

Eukaryotes transport biomolecules between intracellular organelles and between cells and the environment via vesicle trafficking. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNARE proteins) play pivotal roles in vesicle and membrane trafficking. These proteins are categorized as Qa, Qb, Qc, and R SNAREs and form a complex that induces vesicle fusion for targeting of vesicle cargos. As the core components of the SNARE complex, the SNAP25 Qbc SNAREs perform various functions related to cellular homeostasis. The Arabidopsis thaliana SNAP25 homolog AtSNAP33 interacts with Qa and R SNAREs and plays a key role in cytokinesis and in triggering innate immune responses. However, other Arabidopsis SNAP25 homologs, such as AtSNAP29 and AtSNAP30, are not well studied; this includes their localization, interactions, structures, and functions. Here, we discuss three biological functions of plant SNAP25 orthologs in the context of AtSNAP33 and highlight recent findings on SNAP25 orthologs in various plants. We propose future directions for determining the roles of the less well-characterized AtSNAP29 and AtSNAP30 proteins.

• Molecules and Cells
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• v.25 no.1
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• pp.7-19
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• 2008

Complexins play a critical role in the control of fast synchronous neurotransmitter release. They operate by binding to trimeric SNARE complexes consisting of the vesicle protein Synaptobrevin and the plasma membrane proteins Syntaxin and SNAP-25, which are key executors of membrane fusion reactions. SNARE complex binding by Complexins is thought to stabilize and clamp the SNARE complex in a highly fusogenic state, thereby providing a pool of readily releasable synaptic vesicles that can be released quickly and synchronously in response to an action potential and the concomitant increase in intra-synaptic $Ca^{2+}$ levels. Genetic elimination of Complexins from mammalian neurons causes a strong reduction in evoked neurotransmitter release, and altered Complexin expression levels with consequent deficits in synaptic transmission were suggested to contribute to the etiology or pathogenesis of schizophrenia, Huntington's disease, depression, bipolar disorder, Parkinson's disease, Alzheimer's disease, traumatic brain injury, Wernicke's encephalopathy, and fetal alcohol syndrome. In the present review I provide a summary of available data on the role of altered Complexin expression in brain diseases. On aggregate, the available information indicates that altered Complexin expression levels are unlikely to have a causal role in the etiology of the disorders that they have been implicated in, but that they may contribute to the corresponding symptoms.

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.277-284
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• 1993

A possible involvement of maturation promoting factor (nfPF) in the two-cell block phenomenon was studied by fusion experiments. Germinal vesicle (GlF) ooeyte was fused with a blastomore from late or blocked 2-cell mouse embryos. and germinal vesicle breakdoum (GVBD) of fused GV oocvtes in the presence of dbcAMP (100$\mu$g/ml) was scored as an index of MPF aniviD. GnD was induced approximately 30% by fusion of a blastomere derived from late 2-cell embryos, but not from blocked 2-cell embryos. The rate of GVBD was changed when GV oocyte was fused with a blastomere from late 2-cell embryos which were treated with u-amanitin, puromvcin or colcemid before and after hsion: Treatment of late 2-cell embryos with puromycin (50 Is/mll but not with u-amanitin (100 Is/ml) clearly inhibited GVBD, indicating that do novo protein synthesis maw be required for the appearance of MPF activity in late 2-cell embryos. Treatment of late 2-cell embryos w기h colcemid (0.1 Is/mll doubled GVBD, presumably due to the maintenance of metaphase or mitotic phase. SDS-PAGE and twoiimensional electrophoresis revealed that there was no difference in protein synthetic pattern in late and blocked 2-cell embryos, but three phosphoproteins with 27, 35 and 46 M)a, presumsblv M-phase components were phosphorylated in late 2-cell embryos but not in blocked 2-cell embryos. It seems then that MPF activity is closely related to phosphorylstion of M-phase components in late 2-cell embryos.

• Molecules and Cells
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• v.45 no.11
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• pp.806-819
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• 2022

Synaptic accumulation of α-synuclein (α-Syn) oligomers and their interactions with VAMP2 have been reported to be the basis of synaptic dysfunction in Parkinson's disease (PD). α-Syn mutants associated with familial PD have also been known to be capable of interacting with VAMP2, but the exact mechanisms resulting from those interactions to eventual synaptic dysfunction are still unclear. Here, we investigate the effect of α-Syn mutant oligomers comprising A30P, E46K, and A53T on VAMP2-embedded vesicles. Specifically, A30P and A53T oligomers cluster vesicles in the presence of VAMP2, which is a shared mechanism with wild type α-Syn oligomers induced by dopamine. On the other hand, E46K oligomers reduce the membrane mobility of the planar bilayers, as revealed by single-particle tracking, and permeabilize the membranes in the presence of VAMP2. In the absence of VAMP2 interactions, E46K oligomers enlarge vesicles by fusing with one another. Our results clearly demonstrate that α-Syn mutant oligomers have aberrant effects on VAMP2-embedded vesicles and the disruption types are distinct depending on the mutant types. This work may provide one of the possible clues to explain the α-Syn mutant-type dependent pathological heterogeneity of familial PD.

• Development and Reproduction
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• v.1 no.2
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• pp.133-139
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• 1997

To study the function and structure of Golgi apparatus in the spermiogenesis of long-fingered bat (Miniopterus schreibersi fuliginosus), the testis obtained from adult bat was treated with the prolonged osmification or fixed with ferrocyanide reduced osmium. golgi apparatus was oval shape in early Golgi phase, and was composed of cortex and medullar enclosing acrosome in mid Golgi phase. The vesicles of crescent shape Golgi apparatus were closed or fused with small or large vesicles at the periphery of acrosome. Golgi apparatus moved behing the acrosome face in cap phase, but the Golgi apparatus was still active. According to this, Golgi apparatus appears to be involved in the formation of acrosome and sperm tail. Transfer of materials from Golgi to acrosme seems to be carried out not only by fusion of large vesicles with acrosomal vesicles but also by detachment of coated vesicle from various cisternae of Golgi fusing with acrosomal vesicle.

• Korean Journal of Veterinary Research
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• v.22 no.2
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• pp.187-195
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• 1982

This paper dealt with the light microscopical and electron microscopical findings on the morphological changes of the liver of Korean native goat injected with toxin (culture filtrate) of Clostridium perfringens which was isolated from Korean native cattle died of acute Clostridium perfringens enterotoxemia. The results observed are summarized as follows. In the microscopical findings, hyperemia and minute hemorrhage of the liver parenchyma, dilatation of hepatic central vein and centrilobular necrosis of liver, cloudy swelling and hydropic degeneration of hepatic cells, and appearance of light eosinophilic granular bodies in the vacuoles were recognized. In the electron microscopical findings, appearance of pinocytotic vesicle (coated vesicle), fusion of these vesicles, formation of vacuole and accumulation of minute granular proteinous materials in the vacuole were observed in the hepatic cells. Decreased number of glycogen granules, swelling and destruction of mitochondria, proliferation of smooth-surfaced endoplasmic reticulum, enlargement of rough-surfaced endoplasmic reticulum, dispersal of thready agranular membranous structure and appearance of secondary lysosome were recognized in the hepatic cell cytoplasm.