• Title/Summary/Keyword: Golgi apparatus

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Proteomic Changes by Acupuncture Stimulation at HT7 in the Hippocampus of Rat Pups (신문혈 자침이 어린 백서 해마의 단백질 발현에 미치는 영향)

  • Bae, Chang-Hwan;Kim, Dong-Soo;Kim, Seung-Tae
    • Korean Journal of Acupuncture
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    • v.29 no.2
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    • pp.260-270
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    • 2012
  • Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

Change of Fatty Acid Compositions during Hepatic Triacylglycerol Accumulation in Dietary Orotic Acid-induced Fatty Liver (오로토산 유발 지방산의 간장 트리아실글리세롤 축적간 지방산 조성의 변화)

  • 차재영;김경숙;조영수
    • The Korean Journal of Food And Nutrition
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    • v.11 no.5
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    • pp.542-549
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    • 1998
  • Dietary orotic acid is known to induce the fatty liver. Fatty acid profiles in the lipid fraction of the liver and the serum in rats fed with or with orotic acid diet were analyzed. In all the hepatic lipid fraction of rats fed on the supplemented orotic acid diet, there was a significant increased in linoleic acid. In addition, linoleic acid was also increased in the triacylglycerol fraction of hepatic endoplasmic reticulum and the triacylglycerol and diacylglycerol fractions of hepatic Golgi apparatus of the orotic acid-feeding rats. In the time course study of the fatty acid profile in the hepatic triacylglycerol and diacylgycerol fractions, an increase of linoleic acid was observed similarly in the initial stages of orotic acid intake in the both fractions. However, linoleic acid in the serum triacylglycerol fraction of orotic acid-feeding rats increased from day 1, but it began to decrease the increment from day 2, resulting in the lower level of linoleic acid in the serum triacylglycerol fraction of orotic acid-feeding rats than that of rat fed a orotic acid-free diet after 10 days. Oleic acid (18:1) was increased in the only cholesteryl ester fraction of helpatic. However, oleic acid level in other fractions was not changed. The compositions of 14:0, 16:0 and 18:0 was reduced in the hepatic triacylogylcerol, diacylglycerol and cholesteryl ester fractions by orotic acid-feeding. However, these saturated fatty acids were significantly increased in the serum triacylglycerol fractions. The orotic acid indcued changes in linoleic acid level in hepatic triacylglycerol may be explained by the impaired fatty acid metabolism and limited excretion of this fatty acid from liver to serum.

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Evidence of an Alternative Route of Cellobiase Secretion in the Presence of Brefeldin A in the Filamentous Fungus Termitomyces clypeatus

  • Banik, Samudra Prosad;Pal, Swagata;Chowdhury, Sudeshna;Ghorai, Shakuntala;Khowala, Suman
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.412-420
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    • 2011
  • Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

Morphological Study on the Osphradium of Rapana venosa (Gastropoda : Muricidae) (피뿔고동 ( Rapana venosa Valenciennes )의 Osphardium 에 관한 형태학적 연구)

  • 이정재;김성훈
    • The Korean Journal of Malacology
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    • v.4 no.1
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    • pp.1-16
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    • 1988
  • The authors observed histochemical and ultrastructural characters on the osphradium of Rapana venosa Valenciennes using light microscope, scanning and transmission electron microscpes. The results were as follows:1)The basic structure of osphradium was bipectinated shape, which consisted of a septum situating in the center of osphradium and numerous osphradial leaflets. On the other hand, Epidermis of ospradial leaflets formed the structure of pseudostratified ciliated columnar epithelium which was composed of an epithelial cell layer, a basal cel layer and a neuropile. 2) Ciliated dpithelial cells:A large number of these cells were observed on the lateral and ventral regions but a small number of them were observed on the dorsal region. These cells had cylindrical microvilli, slender mitochondria and serve fibers.3) Supporting cells: These cells had cylindrical microvilli, spongy layer, electron dense granules, mitochondria and nerve fibers4) Four types secretory epothelial cells: Four distinct types of secretory epithelial cells were recognized and were arbitrily designated as Type I, Type II, Type III and Type IV.cell type I: These cells contained electron denwe granules(diameter, 0.94-1.56${\mu}{\textrm}{m}$), well developed Golgi apparatus and rough endoplasmic reticula, cell type II: These cills contained two types of granules of the different electron density. One was high electron density granules which were 0.4-1.0${\mu}{\textrm}{m}$ in diameter, The other was low electron density granules which were 0.75-1.2${\mu}{\textrm}{m}$ in diameter.cell type III:These cells had fibrous secretory materials and exhibited strongly positive reaction with Toluidine blue.cell type IV:A large number of this type of cells were observed on the ventral region of ospgradial leaflets and positively reacted with periodic acid Schiff reagent. 5)Dark cells contained several electron dense cillaty rootlets and unmerous granules but cellular organelles were not observed.6) Four types basal cells: Four distinci types of basal cells were recognized and arbitrarily designated as Type I, Type II, Type III and Type IV.Cell type I(light cell): These cells exhibited low electuon density and contained short smooth endoplasmic reticula, several vacuoles and granules.

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Production and identification of antisera against mu-opioid receptor using synthetic peptide epitope (Synthetic peptide를 이용한 mu-opioid receptor에 대한 항혈청의 생산과 검정)

  • Lee, Jang-hern;Kwon, Young-bae;Han, Ho-jae
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.45-54
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    • 1999
  • In the present study we have analyzed the characteristics and distribution of the mu-opioid receptor(MOR) by raising anti-peptide antisera to the C-terminal peptide of MOR. The antisera against MOR was produced in New Zealand White rabbit against 15 residue corresponding to amino acids, 384-398 of the cloned rat MOR. The antigenic peptide was synthesized using an Applied Biosystems 432 solid-phase peptide synthesizer. The specificity and identification of the antisera were tested by analysis of transfected cells, epitope mapping and immunohistochemical method. COS-7 cells electroporated with MOR cDNA were used to evaluate the characteristics and subcellular distribution of MOR. MOR immunoreactivity was prodominent in the plasmalemma and subcellular compartments such as endoplasmic reticulum, Golgi apparatus and vesicle like structure. Furthermore, both tissue sections and transfected cell lines could be immunostained with these antisera and the immunoreactivity was abolished when anti-MOR sera were preincubated with the peptide against which they were raised. Based on epitope mapping analysis, all antisera appeared to have a similar epitope, which was determined to be within the last amino acid, 391-398. Moreover, immunohistochemistry showed that MOR immunoreactivity was observed in many brain areas including cerebral cortex, striatum, hippocampus, locus coeruleus and the superficial laminae of the dorsal horn. These stained spinal cord and brain areas showed the mirrored pattern observed in auto radiographic studies of mu-opioid binding as well as a pattern similar to that seen by is situ hybridization for MOR. Thus, several lines of evidence support the conclusion that the antisera produced in the present study most likely recognize mu-opioid receptor. These results suggest that MOR antisera may be utilized as useful tool to analyze the physiological and pharmacological studies for mu-opioid receptor in the future.

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Fine structural studies on changes of fat bodies in Pieris rapae L. and Bombyx mori. L. during metamorphosis (배추흰나비 (Pieris rapae L.)와 누에나방(Bombyx mori L.) 의 변태(變態)에 따른 지방체(脂肪體)의 미세구조(微細構造) 변화(變化)에 관한 연구(硏究))

  • Han, S.S.;Kim, J.H.;Kim, C.W.;Kim, W.K.
    • Applied Microscopy
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    • v.12 no.2
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    • pp.35-44
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    • 1982
  • The fat bodies of cabbage worm (Pieris rapae) and silk worm (Bomyx mori) during metamorphosis was comparatively studied by electron microscope. 1. Cell oranelles: Golgi apparatus were not observed in both species. It is observed that RER of cabbage worms initiate to degenerate in prepupa stage with complete degeneration at adult stage, while that of silk worms shows similar degenerative pattern. However, mitochondria of cabbage worms are transformed into autophagic vacuole from prepupa stage until adult stage whereas those of silk worm shows a decrease in number in prepupa stage but maintains a certain level until adult stage. 2. Storage substance in cell: Lipid droplets in cabbage worms were observed to increase in numbers during larval stage but afterward decrease in number with an enlargement in size. However immediately after their pupal stage, they almost disappear. On the contrary lipid droplets in silk worms show rather increase in number until adult stage. Protein storage granules in bothspecies were arised from autophagic vacuoles(lysosome) . Fat cells of cabbage worm in adult stage turn out to be residual bodies which last until final stage, but those of silk worm rapidly decrease. Glycogen particles in both species reach maximum at last larval instar and thee gradually decrease thereafter. 3. Fat body sheath: The average width of fat body sheath was measured to be $0.2{\mu}m$ and $0.6{\mu}m$ and surface of fat cells adjacent to fat body sheath in silk worm is heavily infolded.

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Fine Structure of Pericanalicular Cytoplasm of Taurocholic Acid-treated Rat Liver as Revealed by Deep Etching with Rapid Freezing (Taurocholic acid 투여 흰쥐 담세관주위세포질의 미세구조에 관한 급속동결 deep etching법에 의한 연구)

  • Shin, Young-Chul
    • Applied Microscopy
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    • v.28 no.1
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    • pp.73-82
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    • 1998
  • To elucidate how microfilaments and vesicles participate in bile formation, the pericanalicular cytoplasms were observed in the liver of rats treated with taurocholic acid by deep etching with rapid freezing, and copmpared them with the findings on convensional thin sections. The microfilaments were identified around the bile canaliculi in the forms of core filaments of microvilli, filaments of pericanalicular web running in parallel to the border of bile canaliculi, and filaments on the junctional complex. In taurocholic acid-treated rats, microfilaments could be visualized around the bile canaliculi and along their borders. The microfilaments appeared to be installed to link to both the canalicular membrane and vesicles. Such specialized microfilaments are considered to participate in the translocation of vesicles in the pericanalicular cytoplasm. From the evidence, it is assumed that the microfilament induces the vesicles to transport and fuse to bile canalicull into which bile acids is secreted by exocytosis.

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Ultrastructural changes of secretory cells in Harderian gland during postnatal development of Mongolian gerbil (몽고리안 저빌의 출생후 Harderian gland 분비세포 미세구조의 전자현미경적 고찰)

  • Oh, Seung-hyun;Yun, Young-min;Park, Ji-young;Yoon, Yeo-sung;Lee, Joon-sup;Seong, Je-kyung
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.257-265
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    • 2000
  • Harderian glands are the unique organs in several mammals, which human and non-human primates do not have. We report the ultrastructural changes in the postnatal developmental periods of Harderian glands in Mongolian gerbil(Meriones unguiculatus). Male and female Mongolian gerbils were sacrificed on days 3, 10, 30 and 60 after birth and their Harderian glands were observed by transmission electron microscope. The obtained results were summarized as follows; 1. In 3-day-old Mongolian gerbils, Harderian gland was composed of one excretory duct and immature tubules which have two type cells, dark and light cells, identified electron-dense and electron-lucent respectively. 2. In 10-day-old Mongolian gerbils, small lipid vacuoles began to be found in the cytoplasm of the secretory cells of the Harderian gland. Mitochondria, Golgi apparatus, polysomes and slash were more abundant in the cytoplasm of dark cells than those of light cells. The arrangement of tubules in the gland was much more condensed than that of 3-day-old Mongolian gerbils. 3. In 30-day-old Mongolian gerbils, the secretory cells of the tubule were typically columnar in shape and there was one type cell in the tubule. Most of the columnar secretory cells contained various size vacuoles. 4. In 60-day-old Mongolian gerbils, the Harderian gland possessed the typical structural characteristics of adults. The mature glandular structures were more significant than those of 30-day-old animals.

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Free Volume in Polyers Note II。: Positron Annihilation lifetime Spectroscopy and Applications

  • G. Consolati;M. Pegoraro;L. Zanderighi
    • Korean Membrane Journal
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    • v.1 no.1
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    • pp.25-37
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    • 1999
  • positron annihilation Lifetime Spectroscopy has been extensively applied in recent years to investigate the free volume in polymers owing to the capability of the electron-positron bound system (positronium) to probe the typical size of sub-nanometric cavities among the macromolecular chains. In this paper we show recent results obtained through this technique in some amorphous polymeric mem-branes(olyurethanes. PUs and polytrimethilsylilpropine PTMSP) after a brief survey of the general features of the annihilation process as well as of the experimental apparatus. Lifetime of o-ps decay({{{{ tau _3}}}}) in PUs increases going from sub {{{{ TAU _g}}}} to over {{{{ TAU _g}}}} temperatures following a sigmoid curve. The coefficient of dilatation of the free volume fraction is shown to be the sum of two contributes due to the variation with T of the number of holes and of their mean volume. PAL spectrum of PTMSP freshly prepared shows four lifetime components: {{{{ tau _3}}}} and {{{{ tau _4}}}}: only are useful for free volume study. Two kinds of holes of different equivalent radius are reported ({{{{ gamma _s}}}} 4.60 nm and {{{{ gamma _1}}}} 0.754) The equivalent volume does not change in a range of 100 K. however the physical aging increases density and decreases oxygen permeability while {{{{ gamma _s}}}} goes down to 0.374 and r1 to 0.735 The number of holes obtained from the intensities{{{{ IOTA _3}}}} and {{{{ IOTA _4}}}} of PAL spectra decreases with aging 21.7% and 3.5% for large and small holes respectively.

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Expression of Sara2 Human Gene in Erythroid Progenitors

  • Jardim, Denis Leonardo Fontes;Cunha, Anderson Ferreira Da;Duarte, Adriana Da Silva Santos;Santos, Camila Oresco Dos;Saad, Sara Terezinha Olalla;Costa, Fernando Ferreira
    • BMB Reports
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    • v.38 no.3
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    • pp.328-333
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    • 2005
  • A human homologue of Sar1, named Sara2, was shown to be preferentially expressed during erythropoiesis in a culture stimulated by EPO. Previous studies, in yeast, have shown that secretion-associated and Ras-related protein (Sar1p) plays an essential role in protein transport from the endoplasmic reticulum to the Golgi apparatus. Here, we report the molecular analysis of Sara2 in erythroid cell culture. A 1250 bp long cDNA, encoding a 198 amino-acid protein very similar to Sar1 proteins from other organisms, was obtained. Furthermore, we also report a functional study of Sara2 with Real-time quantitative PCR analysis, demonstrating that expression of Sara2 mRNA increases during the initial stages of erythroid differentiation with EPO and that a two-fold increase in expression occurs following the addition of hydroxyurea (HU). In K562 cells, Sara2 mRNA was observed to have a constant expression and the addition of HU also up-regulated the expression in these cells. Our results suggest that Sara2 is an important gene in processes involving proliferation and differentiation and could be valuable for understanding the vesicular transport system during erythropoiesis.