• Title/Summary/Keyword: vesicle

Search Result 742, Processing Time 0.029 seconds

Large Unilamellar Phospholipid Vesicles as a Model Substrate for Phospholipase D

  • Kim Chanwoo;Koh Eun-Hie;Choi Myung-Un
    • Bulletin of the Korean Chemical Society
    • /
    • v.13 no.4
    • /
    • pp.381-384
    • /
    • 1992
  • The hydrolytic susceptibility of large unilamellar vesicle (LUV) toward cabbage phospholipase D (PLD) was studied. The activity of PLD was determined by pH stat titration method. Using phosphatidylcholine LUV as substrate a pH optimum of 6.96 was observed. For maximal activity the optimal temperature of $31^{\circ}C$ and 10 mM of Ca2+ were required. The apparent Km value estimated was 2.5 mM. The hydrolytic activity of PLD toward PC LUV was somewhat high despite the absence of activator in assay system and this high susceptibility of PC LUV may be attributed to the structural properties of LUV. The effect of amphiphatic substances such as dicetyl phosphate and phosphatidic acid on the enzyme activity were also examined in mixed LUVs.

Development of Allotriploid Embryos Produced by Crossing Female Starry Flounder Platichthys stellatus and Male Stone Flounder Kareius bicoloratus (강도다리(Platichthys stellatus, ♀)와 돌가자미(Kareius bicoloratus, ♂) 간 잡종 3배체의 난발생)

  • Jung, Hyo Sun;Lee, Il Yong;Lee, Hyo Bin;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.52 no.3
    • /
    • pp.298-301
    • /
    • 2019
  • We investigated the development of allotriploid embryos derived from a cross between female starry flounder Platichthys stellatus and male stone flounder Kareius bicoloratus. The second cleavage, mid-blastula, gastrula, and Kupffer's vesicle appearance stages, and hatching of embryos began 3.7, 25.6, 45.7, 87.7, and 213.2 h after cold shock at $6^{\circ}C$, respectively. The hatching and development time of triploid interspecific hybrid eggs was approximately the same as those of diploid starry flounder eggs at the same incubation temperature.

Visualization of Extracellular Vesicles of Prokaryotes and Eukaryotic Microbes

  • Kim, Ki Woo
    • Applied Microscopy
    • /
    • v.48 no.4
    • /
    • pp.96-101
    • /
    • 2018
  • The release of nanoscale membrane-bound vesicles is common in all three domains of life. These vesicles are involved in a variety of biological processes such as cell-to-cell communication, horizontal gene transfer, and substrate transport. Prokaryotes including bacteria and archaea release membrane vesicles (MVs) (20 to 400 nm in diameter) into their extracellular milieu. In spite of structural differences in cell envelope, both Gram-positive and negative bacteria produce MVs that contain the cell membrane of each bacterial species. Archaeal MVs characteristically show surface-layer encircling the vesicles. Filamentous fungi and yeasts as eukaryotic microbes produce bilayered exosomes that have varying electron density. Microbes also form intracellular vesicles and minicells that are similar to MVs and exosomes in shape. Electron and fluorescence microscopy could reveal the presence of DNA in MVs and exosomes. Given the biogenesis of extracellular vesicles from the donor cell, in situ high-resolution microscopy can provide insights on the structural mechanisms underlying the formation and release of microbial extracellular vesicles.

Extracellular Vesicles in Psychiatry Research in the Context of RDoC Criteria

  • Ilgin, Can;Topuzoglu, Ahmet
    • Psychiatry investigation
    • /
    • v.15 no.11
    • /
    • pp.1011-1018
    • /
    • 2018
  • The analysis of extracellular vesicles has been accelerated because of the technological advancements in omics methods in recent decades. Extracellular vesicles provide multifaceted information regarding the functional status of the cells. This information would be critical in case of central nervous system cells, which are confined in a relatively sealed biological compartment. This obstacle is more dramatic in psychiatric disorders since their diagnosis primarily depend on the symptoms and signs of the patients. In this paper, we reviewed this rapidly advancing field by discussing definition of extracellular vesicles, their biogenesis and potential use as clinical biomarkers. Then we focused on their potential use in psychiatric disorders in the context of diagnosis and treatment of these disorders. Finally, we tried to combine the RDoC (Research Domain Criteria) with the use of extracellular vesicles in psychiatry research and practice. This review may offer new insights in both basic and translational research focusing on psychiatric disorders.

Antigen Delivery Systems: Past, Present, and Future

  • Hyun-Jeong Ko;Yeon-Jeong Kim
    • Biomolecules & Therapeutics
    • /
    • v.31 no.4
    • /
    • pp.370-387
    • /
    • 2023
  • The COVID-19 pandemic has increased demand for safe and effective vaccines. Research to develop vaccines against diseases including Middle East respiratory syndrome, Ebolavirus, human immunodeficiency virus, and various cancers would also contribute to global well-being. For successful vaccine development, the advancement of technologies such as antigen (Ag) screening, Ag delivery systems and adjuvants, and manufacturing processes is essential. Ag delivery systems are required not only to deliver a sufficient amount of Ag for vaccination, but also to enhance immune response. In addition, Ag types and their delivery systems determine the manufacturing processes of the vaccine product. Here, we analyze the characteristics of various Ag delivery systems: plasmids, viral vectors, bacterial vectors, nanoparticles, self-assembled particles, natural and artificial cells, and extracellular vesicles. This review provides insight into the current vaccine landscape and highlights promising avenues of research for the development and improvement of Ag delivery systems.

Autoradiographic Studies on the Inhibitory Effect of Dibutyryl Cyclic AMP on Mouse Oocyte Maturation in Vitro (Dibutyryl Cyclic AMP가 생쥐여포난자의 성숙에 미치는 억제효과에 관한 자기방사법적 연구)

  • Choi, Choon-Keun
    • Applied Microscopy
    • /
    • v.7 no.1
    • /
    • pp.21-43
    • /
    • 1977
  • This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

  • PDF

Phototransduction and Visual Cycle in the Ascidian Tadpole Larva

  • Kusakabe, Takehiro;Nakashima, Yuki;Kusakabe, Rie;Horie, Takeo;Kawakami, Isao;Yoshida, Reiko;Inada, Kyoko;Nakagawa, Masashi;Tsuda, Motoyuki
    • Journal of Photoscience
    • /
    • v.9 no.2
    • /
    • pp.37-40
    • /
    • 2002
  • Ascidians are lower chordates, and their tadpole-like larvae share a basic body plan with vertebrates. To study photoreceptive systems in ascidians, we have isolated and characterized cDNA clones for three opsins, five G protein ${\alpha}$ subunits (G${\alpha}$), catalytic and regulatory subunits of cGMP phosphodiesterase (PDE), and arrestin from the ascidian Ciona intestinalis tadpole larva. Ci-opsin1 and Ci-opsin2 are vertebrate-type opsins, while Ci-opsin3 is a retinal photoisomerase similar to retinochrome and mammalian RGR. Both Ci-opsin1 and arrestin are specifically localized in the photoreceptor cells of the ocellus, whereas Ci -opsin2 is not expressed in the photoreceptors, but is co-localized in another population of neurons in the brain with PDE (Ci-PDE9 and Ci-PDE$\delta$). Ci-opsin3 is present in the entire region of the brain. Though five different cDNAs encoding Ga have been cloned, no transducin-type G protein has been found yet. Interestingly, one of G${\alpha}$i isoform is conspicuously expressed in the entire region of the brain. The Ci-opsin3 gene expression was observed in a broad area of the brain vesicle as well as in the visceral ganglion. Genes encoding ascidian homologs of CRALBP and ${\beta}$-CD, whose function is required for the mammalian visual cycle, are co-expressed with Ci-opsin3 in the brain vesicle and visceral ganglion. Localization of Ci-opsin3, CRALBP, and ${\beta}$-CD in a broad area of the brain suggests that the brain of the ascidian larva has a visual cycle system similar to that of the vertebrate RPE. Based on these data, we discuss the evolution of vertebrate visual systems.

  • PDF

Mycorrhizal Root Infection and Growth of Cucumber and Tomato Plants by the Inoculated with Glomus sp. In solid Medium Culture (균근균 Glomus sp. 접종에 따른 고형배지경 오이와 방울토마토의 균근 형성과 생육)

  • Cho, Ja-Yong;Kim, Young-Ju;Jin, Seo-Young;Kang, Sung-Gu;Kim, Hong-Lim;Sohn, Bo-Kyoon
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.37 no.5
    • /
    • pp.341-349
    • /
    • 2004
  • This study was conducted to compare the effects of arbuscular mycorrhizal fungi (AMF) on the growth and fruit yield of hydroponically grown cucumber and tomato plants in solid medium culture. Mycorrhizal fungus Glomus sp. was collected from plastic film house soils of cucumber and tomato and inoculated to the experimental crops at the time of seeding and transplanting. Root infection of cucumber and tomato plants by AMF was more significantly increased when the AMF was inoculated at seeding stage than at transplanting stage. In the infected roots of cucumber and tomato, mycorrhizal hyphae was easily observed but vesicle and arbuscule were rare. Overall plant growth was increased with AMF inoculation and the growth was higher when AMF was inoculated at seeding stage. Fresh weight of each fruit of cucumber and tomato and sugar content in tomato fruits were significantly increased with AMF inoculation at seeding stage. The AMF inoculation also increased fruit yields of cucumber and tomato.

Effects of Barbiturates on the Fluidity of Phosphatidylethanolamine Model Membranes (Barbiturates가 소의 신선한 대뇌피질 Synaptosomal Plasma Membrane Vesicles로 부터 추출하여 제제한 Phosphatidylethanolamine 인공세포막의 유동성에 미치는 영향)

  • Yun, Il;Kim, Hyung-Il;Hwang, Tae-Ho;Kim, Jong-Ryol;Kim, In-Se;Chung, Yong-Za;Shin, Yong-Hee;Jung, Hyun-Ok;Kang, Jung-Sook
    • The Korean Journal of Pharmacology
    • /
    • v.26 no.2
    • /
    • pp.209-217
    • /
    • 1990
  • Intramolecular excimer formation with 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization with 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effects of barbiturates on the bulk fluidity of the model membranes of phosphatidylethanolamine fraction of synaptosomal plasma membrane vesicles (SPMVPE) isolated from bovine cerebral cortex. In the SPMVPE, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the fluorescence polarization (P), anisotropy (r), limiting anisotropy $(r_{8})$, order parameter (S) and rotational relaxation time $({\bar{P}})$ of DPH in a dose-dependent manner. The relative potencies of barbiturates to order the SPMVPE were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. Hence, it is concluded that barbiturates have ordering effects on the SPMVPE. And the membrane-ordering potencies of barbiturates appear to be correlated with the potencies for enhancement of GABA-stimulated chloride influx and with the anesthetic effects of barbiturates.

  • PDF

Subcellular Characterization of Porcine Oocytes with Different Glucose-6-phosphate Dehydrogenase Activities

  • Fu, Bo;Ren, Liang;Liu, Di;Ma, Jian-Zhang;An, Tie-Zhu;Yang, Xiu-Qin;Ma, Hong;Zhang, Dong-Jie;Guo, Zhen-Hua;Guo, Yun-Yun;Zhu, Meng;Bai, Jing
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.28 no.12
    • /
    • pp.1703-1712
    • /
    • 2015
  • The in vitro maturation (IVM) efficiency of porcine embryos is still low because of poor oocyte quality. Although brilliant cresyl blue positive (BCB+) oocytes with low glucose-6-phosphate dehydrogenase (G6PDH) activity have shown superior quality than BCB negative (-) oocytes with high G6PDH activity, the use of a BCB staining test before IVM is still controversial. This study aimed to shed more light on the subcellular characteristics of porcine oocytes after selection using BCB staining. We assessed germinal vesicle chromatin configuration, cortical granule (CG) migration, mitochondrial distribution, the levels of acetylated lysine 9 of histone H3 (AcH3K9) and nuclear apoptosis features to investigate the correlation between G6PDH activity and these developmentally related features. A pattern of chromatin surrounding the nucleoli was seen in 53.0% of BCB+ oocytes and 77.6% of BCB+ oocytes showed peripherally distributed CGs. After IVM, 48.7% of BCB+ oocytes had a diffused mitochondrial distribution pattern. However, there were no significant differences in the levels of AcH3K9 in the nuclei of blastocysts derived from BCB+ and BCB- oocytes; at the same time, we observed a similar incidence of apoptosis in the BCB+ and control groups. Although this study indicated that G6PDH activity in porcine oocytes was correlated with several subcellular characteristics such as germinal vesicle chromatin configuration, CG migration and mitochondrial distribution, other features such as AcH3K9 level and nuclear apoptotic features were not associated with G6PDH activity and did not validate the BCB staining test. In using this test for selecting porcine oocytes, subcellular characteristics such as the AcH3K9 level and apoptotic nuclear features should also be considered. Adding histone deacetylase inhibitors or apoptosis inhibitors into the culture medium used might improve the efficiency of IVM of BCB+ oocytes.