Kim, Seong-Hoon;Lee, Chi-Hoon;Song, Young-Bo;Ju, Hea-Sung;Kim, Hyung-Bae;Lee, Young-Don
Applied Microscopy
/
v.42
no.2
/
pp.61-66
/
2012
Ultrastructural characteristics of blacktip grouper, Epinephelus fasciatus spermatozoa were investigated using transmission and scanning electron microscopy. The spermatozoa of E. fasciatus consisted of a spherical head part, a midpiece with cytoplasmic canal entrance and a flagellum with lateral fins. Internal ultrastructurally, the nucleus contains high electron dense chromatin having granular particles and has no acrosome. The centriolar complex lies outside of the nuclear fossa and it is connected by the osmophilic filaments. Also the osmophilic filaments connect between the centriolar complex and the nuclear membrane. The midpiece contains eight to nine spherical mitochondria, cytoplasmic canal and necklaces. The flagellum has a typical 9+2 axonemal structure. The lateral fins contain vesicles and a typical 9+2 axonemal structure. Consequently this study contributes to comparative grouper spermatology and provide useful systematic taxonomic characters.
Insulin stimulates the fusion of intracellular vesicles containing glucose transporter 4 (GLUT4) with plasma membrane in adipocytes and muscle cells. Here we show that adipocyte differentiation results in enhanced insulin sensitivity of glucose uptake. On the other hand, glucose uptake in response to platelet-derived growth factor (PDGF) stimulation was markedly reduced by adipocyte differentiation. Expression level of insulin receptor and caveolin-1 was dramatically increased during adipocyte differentiation. Adipocyte differentiation caused :ilightly enhanced activation of acutely transforming retrovirus AKT8 in rodent T cell lymphoma (Akt) by insulin stimulation. However, activation of Akt by PDGF stimulation was largely reduced. Activation of ERK was not detected in both fibroblasts and adipocytes after stimulation with insulin. PDGF-dependent activation of ERK was reduced by adipocyte differentiation. Insulin-dependent glucose uptake was abrogated by LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, in both fibroblasts and adipocytes. Also disassembly of caveolae structure by $methyl-\beta-cyclodextrin$ caused impairment of Akt activation and glucose uptake. Finally, insulin receptor, Akt, SH2-domain-containing inositol 5-phosphatase 2 (SHIP2), and regulatory subunit of PI3K are localized at lipid raft domain and the translocation was facilitated upon insulin stimulation. Given these results, we suggest that lipid raft provide proper site for insulin action for glucose uptake.
Cho, Kwang Keun;Lee, Seung Ho;Choi, In Soon;Lee, Sang Won
Journal of Life Science
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v.31
no.6
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pp.595-602
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2021
Human intestinal microbiota play an important role in the regulation of the host's metabolism. There is a close pathological and physiological interaction between dysbiosis of the intestinal microflora and obesity and metabolic syndrome. Akkermansia muciniphila, which was recently isolated from human feces, accounts for about 1-4% of the intestinal microbiota population. The use of A. muciniphila- derived external membrane protein Amuc_1100 and extracellular vesicles (EVs) could be a new strategy for the treatment of obesity. A. muciniphila is considered a next-generation probiotic (NGP) for the treatment of metabolic disorders, such as obesity. Faecalibacterium prausnitzii accounts for about 5% of the intestinal microbiota population in healthy adults and is an indicator of gut health. F. prausnitzii is a butyrate-producing bacterium, with anti-inflammatory effects, and is considered an NGP for the treatment of immune diseases and diabetes. Postbiotics are complex mixtures of metabolites contained in the cell supernatant secreted by probiotics. Parabiotics are microbial cells in which probiotics are inactivated. Paraprobiotics and postbiotics have many advantages over probiotics, such as clear chemical structures, safe dose parameters, and a long shelf life. Thus, they have the potential to replace probiotics. The most natural strategy to restore the imbalance of the intestinal ecosystem normally is to use NGPs among commensal bacteria in the gut. Therefore, it is necessary to develop new foods or drugs such as parabiotics and postbiotics using NGPs.
Purpose: Exosomes are membrane vesicles that are present in body fluids and contain proteins, lipids, and microRNA (miRNA). Periodontal tissue examinations assess the degree of periodontal tissue destruction according to the probing depth (PD), clinical attachment loss (CAL), bleeding on probing, and X-ray examinations. However, the accurate evaluation of the prognosis of periodontitis is limited. In this study, we collected saliva from patients before and after initial periodontal therapy (IPT) and compared changes in the clinical parameters of periodontitis with changes in the components of salivary exosomes. Methods: Saliva was collected from patients with stage III and IV periodontitis at the first visit and post-IPT. Exosomes were purified from the saliva, and total protein and RNA were extracted. Changes in expression levels of C6, CD81, TSG101, HSP70, and 6 kinds of miRNA were analyzed by western blots and real-time polymerase chain reaction. Results: Patients with increased C6 expression after IPT had significantly higher levels of periodontal inflamed surface area (PISA), miR-142, and miR-144 before and after IPT than patients with decreased C6 expression after IPT. Patients with decreased and unchanged CD81 expression after IPT showed significantly higher PD, CAL, and PISA before IPT than after IPT. Patients with decreased and unchanged TSG101 expression after IPT had significantly higher PD before IPT than after IPT. Patients with increased HSP70 expression after IPT had significantly higher PD and PISA before and after IPT than patients with unchanged HSP70 after IPT. The expression levels of miR-142, miR-144, miR-200b, and miR-223 changed with changes in the levels of C6, CD81, TSG101, and HSP70 in the salivary exosomes of periodontitis patients before and after IPT. Conclusions: The expression levels of proteins and miRNAs in salivary exosomes significantly changed after IPT in periodontitis patients, suggesting that the components of exosomes could serve as biomarkers for periodontitis.
The vesicle system of DPPC(dipalmitoylphosphaticylcholine)/Chol(Cholesterol) has been modified by incorporating various mole fractions of flourinated surfactant($C_8F_{17}(CH_2)_2OCO-CH_2CH(SO_3Na)COO(CH_2)_2C_8F_{17}$. Sodium bis(1H,1H,2H,2H-heptadecaflurododecyl)-2-sulfosuccinate, FS)/fluorinated fatty acid salt ($C_7F_{15}COONH_4$, ammoniumpentadecaflurooctyrate, FFS), and their physicochemical properties have been investigated in an attempt to enhance the stability of phospholipid vesicle system. The ${\zeta}$-potential measurement by use of Zetamaster sub-micron Particle Electrophoresis Analyzer (Malvern Co.) showed that a charged homogeneous DPPC/Chol/FS vesicle has been formed owing to the incorporated FFS effect on the membrane, playing a role as a cosurfactant in the bilayer between DPPC and FS components. With increase in the concentration of FFS, it was found that the particle size and also surface charge of the DPPC/Chol/FS vesicle decreased. The stability of DPPC/Chol/FS/FFS liposome was found to be enhanced significantly compared to that of DPPC/Chol/FS according to the dispersity change as a function of time. The release rate of dye molecule of Methylene Blue from the DPPC/Chol/FS/FFS vesicle was determined to be slower than that of DPPC/Chol/FS system, and it may be attributed to the increase in microviscosity of the hydrophobic region in the bilayer. The affinfinity of DPPC/Chol/FS/FFS vesicles to albumin was found to be slightly lowered compared to that of DPPC/Chol/FS. Based on these findings, it was confirmed that a more stable and homogeneous vesicle system of DPPC/Chol/FS could be prepared by addition of FFS, acting as a cosurfactant in the aggregate formation.
The lipids of milk provide energy and many essential nutrients for the newborn animal. They also have distinctive physical properties that affect the processing of dairy products. Milk fat globules mainly consist of neutral lipids like triacylglycerols, whereas the globule membranes contain the complex lipids mostly, Phospholipids are a small but important fraction of the milk lipids and are found mainly in the milk fat globule membrane and other membranous material in the skim-milk phase. The milk fats of ruminant animals are characterized by the presence of relatively high concentrations of short-chain fatty acids, especially butyric and hexanoic acids, which are rarely found in milks of non-ruminants. The fatty acids of milk lipids arise from de novo synthesis in the mammary gland and uptake from the circulating blood. The fatty acid compositions of milks are usually complex and distinctive, depending on the nature of the fatty acids synthesized de novo in the mammary gland and those received from the diet in each species. The content and composition of milks from different species vary widely; presumably, these are evolutionary adaptations to differing environments. The actual process by which these globules are formed is unkonwn, but there are indications that triglyceride-containing vesicles which bleb from endoplasmic reticulum may serve as nucleation sites for globules. Recent studies on milk have centred on the manipulation of milk lipids to increase specific fatty acids, i.e. 20-carbon omega-3 fatty acids (eicosapentaenoic acid 20:5n3, decosahexaenoic acid 22:6n3) from marine sources because the fatty acids are closely associated with a decreased risk of coronary heart disease.
The histochemical characteristics and ultrastructure of the mantle in the spiny top shell, Batillus cornutus were described using light and electron microscopy. The simple epidermal layer wrapped on the top and bottom of the centrally located connective tissue. And then the epidermal layer were divided into the outer epidermal layer near a shell and the inner epidermal layer closed to the visceral mass. The connective tissue layer was composed of the collagen fiber muscularfiber bundle and hemolymph sinus. Mucous cells in the apical mantle contained acid and neutral mucopolysaccaride, and acidic carboxylated mucopolysaccaride in the mid and marginal mantle. The mantle thickness, epidermal layer thickness and hemolymph sinus area displayed a trend of reduction from the marginal zone to the apical zone. From TEM observation, it was possible to distinguish epithelium, ciliated cell, absorptive cell and secretory cell in the epidermal layer. The epithelia were columnar and the nucleus was elliptical. The free surface were covered with microvilli. The lateral membranes of epithelium was con nected with neighboring cells by the zonular occludens, zonular adherens and membrane interdigitation. Ciliated cell on free surface had cilia and microvilli, and numerous mitochondria in the apical cytoplasm. In the epidermal layer, it observed 2 type cells having absorptive function. The absorptive cells were columnar in shape, and contained microvilli, pinocytotic vesicles, mitochondria and lysosomes of various electron density. Secretory cells can be divided into four types (A, B, C, D) depending on the cell shape and characteristics of secretory granules. These cells were unicellular glands and had similar characteristics to previously reported on the mantle of the gastropod and bivalves.
Kim, Woo-Kap;Park, Hong-Duok;Kim, Eun-Soo;Han, Sung-Sik
Applied Microscopy
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v.9
no.1
/
pp.57-69
/
1979
The ultrastructural changes of embryo and endosperm cells were observed during the green fruit with embryo about $250{\mu}$ long to germination. 1. In the embryo cells of green fruit with embryo about $250{\mu}$ long, mitochondrial cristae and plastid are undifferentiated and dictyosome are occasionally observed. There are electron-opaque globoids in the vacuole and a lot of spherosomes in the outer layer of smooth endoplasmic reticulum. Endosperm is filled with spherosomes and electron-opaque protein bodies surrounded by spherosomes, and due to these, other organelle are not observed. 2. In the embryo cells of seeds with red seed coat, mitochondrial cristae are well developed, electron-opaque globoids increased, and vacuoles are enlarged. In the endosperm, however, spherosomes increased, protein bodies are enlarged, and electron-opaque globoidal crystals are dispersed within them. 3. In the procambium and epicotyl cells of dehiscent seed, Golgi vacuoles and vesicles are well developed, and mitochondrial cristae are also well differentiated. Spherosomes are numerously present and radicle cells, peripheral cells of hypocotyl, and vacuoles of cotyledon are well differentiated. Endosperm is filled with spherosomes containing electron-opaque granules and protein bodies are surrounded by a single membrane. There are acid phosphatase around globoids and spherosomes. 4. At the time of seeding, spherosomes markedly increased in the outer layer of cotyledon and protein bodies are also observed. Cell organelles are differentiated and plastids containing starch are also present. 5. In the outer $2{\sim}3$ layers of cotyledons, radicle cells, and peripheral cells of hypocotyl during post-seeding to germination, spherosomes and plastids with starch increased, and mitochondria and microbodies are also found around the nucleus of embryo cells. With approaching, the germination stage, in the endosperm contacting with embryo, vacuoles are well differentiated but spherosomes decreased. There increased electron-opaque materials within vacuoles. In other endosperm, with the decrease of spherosome, mitochondria increased and electro n-opaque globular bodies are formed and gradually increased. The outer layer of protein bodies are reduced while electron-transparent portions are enlarged and fused together to occupy the outer layer where small particles are formed. 6. In the endosperm of germination stage, spherosomes decreased while protein bodies, are fused together to form 2 or 3 within a cell.
It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.
In this paper, five kinds of neurosecretory cells-light green (LG) cell, dark green (DG) cell, caudo-dorsal (CD) cell, blue green (BG) cell, and yellow (Y) cell- and neuropils in the cerebral ganglion of the African giant snail, Achatina fulica, were observed with an electron microscope. The following results were obtained. The LG cells are circular or ovoid in shape, and about $60{\mu}m$ in size. The nucleus and cytoplasm of the LG cell look light due to their electron-low density. Large granular chromatins are evenly developed in the karyolymph, where round nucleoli are also found. In the cytoplasm, electron -high dense round granules of $0.4{\mu}m$ in average size are crowded. The DG cells are ovoid in shape, and $50\sim20{\mu}m$ in size. These relatively electron-high dense cells were rarely found. In their cytoplasm, cell organelles such as rough endoplasmic reticulum and mitochondria are found together with electron -high dense round granules of $0.2{\mu}m$ in average size. The CD cells are ellipsoidal cells densely distributed in caudo-dorsal parts of the cerebral ganglion. They have large nuclei compared with the cytoplasm. The developed granular heterochromatins are observed in the karyolymph, and lots of small round granules of $0.12{\mu}m$ in average size in the cytoplasm. The 3G cells, rarely found around endoneurium of the cerebral ganglion, take the shapes of long ellipses. They look dark due to their electron -high density. In the cytoplasm, small round granules of $0.1{\mu}m$ in average size are found. The Y cells are the smallest among the neurosecretory cells($9\times6.6{\mu}m$ in size). They are found mostly between the medio-dorsal parts and the caudo-dorsal parts of the cerebral ganglion. In the cytoplasm, tiny round granules of $0.08{\mu}m$ in average size form a group. The neuropils are found in the middle of the cerebral ganglion. In the axon ending, round granules with electron -high density ($0.07\sim0.03{\mu}m$ in diameter) and lucent vesicles ($0.03{\mu}m$ in diameter) are found in large quantities. They are excreted in the state of exocytosome formed by the invagination of the limiting membrane of the axon ending.
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