• 제목/요약/키워드: vesicle

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Physiological Functions of the COPI Complex in Higher Plants

  • Ahn, Hee-Kyung;Kang, Yong Won;Lim, Hye Min;Hwang, Inhwan;Pai, Hyun-Sook
    • Molecules and Cells
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    • 제38권10호
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    • pp.866-875
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    • 2015
  • COPI vesicles are essential to the retrograde transport of proteins in the early secretory pathway. The COPI coatomer complex consists of seven subunits, termed ${\alpha}-$, ${\beta}-$, ${\beta}^{\prime}-$, ${\gamma}-$, ${\delta}-$, ${\varepsilon}-$, and ${\zeta}$-COP, in yeast and mammals. Plant genomes have homologs of these subunits, but the essentiality of their cellular functions has hampered the functional characterization of the subunit genes in plants. Here we have employed virus-induced gene silencing (VIGS) and dexamethasone (DEX)-inducible RNAi of the COPI subunit genes to study the in vivo functions of the COPI coatomer complex in plants. The ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP subunits localized to the Golgi as GFP-fusion proteins and interacted with each other in the Golgi. Silencing of ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP by VIGS resulted in growth arrest and acute plant death in Nicotiana benthamiana, with the affected leaf cells exhibiting morphological markers of programmed cell death. Depletion of the COPI subunits resulted in disruption of the Golgi structure and accumulation of autolysosome-like structures in earlier stages of gene silencing. In tobacco BY-2 cells, DEX-inducible RNAi of ${\beta}^{\prime}$-COP caused aberrant cell plate formation during cytokinesis. Collectively, these results suggest that COPI vesicles are essential to plant growth and survival by maintaining the Golgi apparatus and modulating cell plate formation.

Overexpression of GmAKR1, a Stress-Induced Aldo/keto Reductase from Soybean, Retards Nodule Development

  • Hur, Yoon-Sun;Shin, Ki-Hye;Kim, Sunghan;Nam, Kyoung Hee;Lee, Myeong-Sok;Chun, Jong-Yoon;Cheon, Choong-Ill
    • Molecules and Cells
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    • 제27권2호
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    • pp.217-223
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    • 2009
  • Development of symbiotic root nodules in legumes involves the induction and repression of numerous genes in conjunction with changes in the level of phytohormones. We have isolated several genes that exhibit differential expression patterns during the development of soybean nodules. One of such genes, which were repressed in mature nodules, was identified as a putative aldo/keto reductase and thus named Glycine max aldo/keto reductase 1 (GmAKR1). GmAKR1 appears to be a close relative of a yeast aldo/keto reductase YakC whose in vivo substrate has not been identified yet. The expression of GmAKR1 in soybean showed a root-specific expression pattern and inducibility by a synthetic auxin analogue 2,4-D, which appeared to be corroborated by presence of the root-specific element and the stress-response element in the promoter region. In addition, constitutive overexpression of GmAKR1 in transgenic soybean hairy roots inhibited nodule development, which suggests that it plays a negative role in the regulation of nodule development. One of the Arabidopsis orthologues of GmAKR1 is the ARF-GAP domain 2 protein, which is a potential negative regulator of vesicle trafficking; therefore GmAKR1 may have a similar function in the roots and nodules of legume plants.

Hershberger Assays for Bisphenol-A and Its Substitute Candidates

  • Kim, Hee-Su;Kim, Yong-Bin;Choi, Donchan;Cheon, Yong-Pil;Lee, Sung-Ho
    • 한국발생생물학회지:발생과생식
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    • 제21권4호
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    • pp.441-448
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    • 2017
  • Bisphenol-A(BPA) is a member of alkylphenol family, and shows adverse effects including reduced fertility, reproductive tract abnormalities, metabolic disorder, cancer induction, neurotoxicity and immunotoxicity. In the present study, we conducted Hershberger assay to evaluate whether the two candidates to replace BPA have androgenic or antiandrogenic activity. The assay was carried out using immature castrated Sprague-Dawley male rats. After 7 days of the surgery, testosterone propionate (TP, 0.4 mg/kg/day) and test materials (low dose, 40 mg/kg/day; high dose, 400 mg/kg/day) were administered for 10 consecutive days by subcutaneous (s.c.) injection and oral gavage, respectively. Test materials were BPA, isosorbide (ISO) and cyclohexanedimethanol (CHDM). The rats were necropsied, and then the weights of five androgen-dependent tissues [ventral prostate, seminal vesicle, levator ani-bulbocavernosus (LABC) muscle, paired Cowper's glands, and glans penis] and three androgen-insensitive tissues (kidney, spleen and liver) were measured. All test materials including BPA did not exhibit any androgenic activity in the assay. On the contrary, antiandrogen-like activities were found in all test groups, and the order of the intensity was CHDM > BPA > ISO in the five androgen-sensitive tissues. There was no statistical difference between low dose treatment and high dose treatment of BPA group as well as ISO group. In CHDM group, high dose treatment exhibited most severe weight reduction in all measured tissues. There was no statistical difference in androgen-insensitive tissue measurements, except BPA groups. Since the effects of ISO treatment on the accessory sex organs were much less or not present at all when compared to those of BPA, ISO could be a strong candidate to replace BPA. CHDM treatment brought most severe weight reduction in all of androgen-sensitive tissues, so this material should be excluded for further screening of BPA substitute selection.

Hershberger Assays for Di-2-ethylhexyl Phthalate and Its Substitute Candidates

  • Kim, Hee-Su;Cheon, Yong-Pil;Lee, Sung-Ho
    • 한국발생생물학회지:발생과생식
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    • 제22권1호
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    • pp.19-27
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    • 2018
  • In the present study, we employed Hershberger assay to determine possible androgenic or antiandrogenic activities of three di-2-ethylhexyl phthalate (DEHP) substitute candidates. The assay was carried out using immature castrated Sprague-Dawley male rats. After 7 days of the surgery, testosterone propionate (TP, 0.4 mg/kg/day) and test materials (low dose, 40 mg/kg/day; high dose, 400 mg/kg/day) were administered for 10 consecutive days by subcutaneous (s.c.) injection and oral gavage, respectively. Test materials were DEHP, 2-ethylhexyl oleate (IOO), 2-ethylhexyl stearate (IOS) and triethyl 2-acetylcitrate (ATEC). The rats were necropsied, and then the weights of five androgen-dependent tissues [ventral prostate, seminal vesicle, coagulating glands, levator ani-bulbocavernosus (LABC) muscle, paired Cowper's glands, and glans penis] and four androgen-insensitive tissues (kidney, adrenal glands, spleen and liver) were measured. All test materials including DEHP did not exhibit any androgenic activity in the assay. On the contrary, antiandrogen-like activities were found in all test groups, and the order of the intensity was ATEC < DEHP < ISO < IOO in the five androgen-sensitive tissues. There was no statistical difference between low dose treatment and high dose treatment of all replacement candidate groups. In DEHP groups, high dose treatment exhibited significant weight gains in LABC and Glan Penis. There was no statistical difference in androgen-insensitive tissue measurements. Since the effects of ATEC treatment on the accessory sex organs were much less or not present at all when compared to those of DEHP, ATEC could be a strong candidate to replace DEHP. IOO treatment brought most severe weight reduction in all of androgen-sensitive tissues, so this material should be excluded for further screening of DEHP substitute selection.

오리나무 열수 추출물이 실험적으로 유발된 흰쥐의 위염 및 위궤양에 미치는 영향 (Effect of Hot Water Extract of Alnus japonica Steud on the Experimentally-induced Acute Gastritis and Peptic Ulcers in Rats)

  • 나천수;이상범;김진범;정하숙;동미숙
    • 생약학회지
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    • 제43권1호
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    • pp.72-78
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    • 2012
  • Alnus japonica Steud (A. japonica) have long been used in the traditional medicine for gastric disorder, hepatitis and fatty liver in Korea. Antiulcer effects of A. japonica hot water extract (AJ ext) were evaluated by in vitro antibacterial activity against H. pylori, by the inhibitory action against the in vitro gastric $H^+/K^+$-ATPase and using rat models of gastric mucosal damage and gastric ulcer induced by HCl-ethanol, indomethacin, and restraint and water-immersion stress. For the determination of antibacterial activity of AJ ext against H. pylori, the activity of urease which released from H. pylori was measured in culture. AJ ext showed weak antibacterial activity against H. pylori with the growth inhibitions of 37% and 61% by adding final concentrations of 500 and $1000{\mu}g/ml$ culture, respectively at 24 h. To observe the inhibitory activity of AJ ext against the $H^+/K^+$-ATPase in hog gastric membrane vesicle, $IC_{50}$ value of AJ ext was $806.3{\mu}g/ml$. Pretreatment of AJ ext (200, 500 mg/kg, p.o.) prevented in a dose-dependent manner the acute gastritis in HCl-ethanol model and the formation of gastric ulcer in indomethacin model and restraint and water-immersion stress model. These results suggest that the AJ ext can be used for prevention and treatment of gastric mucosal damage and ulcers induced by various stress.

Parvatrema chaii n. sp. (Digenea: Gymnophallidae) from mice experimentally infected with metacercariae collected from surf-clam, Mactra veneriformis

  • Sohn, Woon-Mok;Na, Byoung-Kuk;Ryang, Yong-Suk;Ching, Hilda Lei;Lee, Soon-Hyung
    • Parasites, Hosts and Diseases
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    • 제45권2호
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    • pp.115-120
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    • 2007
  • Parvatrema chaii n. sp. (Digenea: Gymnophallidae) is described using the worms recovered from experimentally infected mice in Korea. The metacercariae were collected from surf-clams, Mactra veneriformis, from a tidal flat in Sochon-gun, Chungchongnam-do. The metacercariae were elliptical ($0.262{\times}0.132 mm$), and the genital pore had an anterior arch of 16-17 sensory papillae in scanning electron microscopic view, Adult worms were ovoid to foliate (0.275-0.303 by 0.140-0.150 mm), and their characteristic features included the presence of lateral lips, short esophagus, genital pore located some distance anterior to the ventral sucker, club-shaped seminal vesicle, a compact to slightly lobed vitellarium, elliptical eggs (0.018-0.020 by 0.010-0.013 mm), and absence of the ventral pit. This gymnophallid is classified as a member of the genus Parvatrema because of the location of the wide genital pore some distance from the ventral sucker, and the absence of the ventral pit. It differs from previously reported Parvatrema species, including the type species, P. borinquenae. In particular, the morphologies of the vitellarium and the genital pore with an anterior arch of 16-17 sensory papillae are unique features. Therefore, we propose it as a new species, Parvatrema chaii n. sp. (Digenea: Gymnophallidae).

Influences of Hinge Region of a Systhetic Antimicrobial Peptide, Cecropin A(1-13)-Melittin(1-13) Hybrid on Antibiotic Activity

  • 신송엽;강주현;이동건;장소윤;서무열;김길룡;함경수
    • Bulletin of the Korean Chemical Society
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    • 제20권9호
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    • pp.1078-1084
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    • 1999
  • A synthetic cecropin A(1-13)-melittin(1-13) [CA-ME] hybrid peptide was known to be an antimicrobial peptide having strong antibacterial, antifungal and antitumor activity with minimal cytotoxic effect against human erythrocyte. Analogues were synthesized to investigate the influences of the flexible hinge region of CA-ME on the antibiotic activity. Antibiotic activity of the peptides was measured by the growth inhibition against bac-terial, fungal and tumor cells and vesicle-aggregating or disrupting activity. The deletion of Gln-Gly-Ile (P1) or Gly-Gln-Gly-Ile-Gly (P3) from CA-ME brought about a significant decrease on the antibiotic activities. In contrast, Gly-Ile-Gly deletion (P2) from CA-ME or Pro insertion (P5) instead of Gly-Gln-Gly-Ile-Gly of CA-ME retained antibiotic activity. This result indicated that the flexible hinge or β-bend structure provided by Gly-Gln-Gly-Ile-Gly, Gln-Gly, or Pro in the central region of the peptides is requisite for its effective antibiotic activity and may facilitate easily the hydrophobic C-terminal region of the peptide to penetrate the lipid bilayers of the target cell membrane. In contrast, P4 and P6 with Gly-Gln-Gly-Pro-Gly or Gly-Gln-Pro in the central region of the peptide caused a drastic reduction on the antibiotic activities. This result suggested that the con-secutive β-bend structure provided by Gly-Gln-Gly-Pro-Gly or Gly-Gln-Pro in the central hinge region of the peptide seems to interrupt the ion channel/pore formation on the target cell membranes.

Adverse Effect of Nonylphenol on the Reproductive System in F1 Male Mice: A Subchronic Low-Dose Exposure Model

  • Kim, Yong-Bin;Cheon, Yong-Pil;Lee, Sung-Ho
    • 한국발생생물학회지:발생과생식
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    • 제23권2호
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    • pp.93-99
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    • 2019
  • Nonylphenols (NPs) are widely used industrial materials, and are considered as potent endocrine disrupting chemical. Present study was undertaken to clarify the effect of subchronic low-dose NP exposure to F1 generation male mice. Mice were divided into 2 groups; (1) CON, control animals and (2) NP-50 ($50{\mu}g/L$), animals were treated with NP via drinking water. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 55 of F1 offsprings. Mice were sacrificed on PND 55 and the tissue weights were measured. The initial body weights (at PND 21) and terminal body weights (PND 55) of the NP-50 animals were significantly lower than those of control animals (p<0.05). NP exposure induced a significant increase in the absolute weight of the testes (p<0.05). Conversely, the NP exposure caused significant decrease in the absolute weights of the epididymis (p<0.01), prostate (p<0.05) and seminal vesicle (p<0.05). Histopathological studies revealed that NP-treated animals exerted decreased seminiferous tubule diameters, reduced luminal area, and lower number of germ cells. Also some sloughing morphologies in the tubules were observed. In the caudal epididymis, fewer mature sperms and swollen epithelial cells were found in the NP-treated group. Our results confirmed that the subchronic low-dose NP exposure altered some male parameters and induced histopathological abnormalities in testis and epididymis of F1 mice. Since the NP dose used in this study is close to the average human daily NP exposure, our results could provide practically meaningful understanding of adverse effect of EDC in human.

소금쟁이의 尖體形成 (Acrosome Morphogenesis in Gerris paludum (Heteroptera))

  • Lee, Young-Hwan;Lee, Chang-Eon
    • 한국동물학회지
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    • 제24권2호
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    • pp.65-75
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    • 1981
  • Gerris paludum의 精子形成 동안의 尖體形成에 대한 연구를 요약하면 다음과 같다. 1. Golgi體는 精母細胞의 초기단계에서 細胞質에 산재되어 있다가 서로 융합하여 囊 胚形을 하며 이들이 모여서 감수분열 전기에는 몇 개의 큰 Golgi體群을 형성하는데, 이들은 결국 精細胞에 균등히 분포된다. 2. 精子完成에서 acroblast는 처음에 하나의 큰 胞로 나타나다가, 그 내부에서 尖體顆粒이 分化된 후, acroblast는 顆粒에서 分離되어 결국 tail filament를 따라 사라진다. 3. 尖體는 mitochondrial derivatives의 반대편인 核 전면부로 이동한 후, two zones 즉 core와 sheath로 分化된다. 4. Basal bodies와 tip은 모두 sheath에서 由來한다. Basal bodies는 sheath의 基部에서 발생해서 尖體가 伸長되고 좁아짐에 따라 점차로 尖體의 基部를 싸기 시작하여 결국 완전히 둘러싸게 되며, 分化된 tip은 core의 前端部에 인접해서 나타나지만 sheath와 뚜렷이 연결되어 있다. 5. 分化된 tip은 basal bodies보다 앞서서 伸長한다. Basal bodies는 精細胞 후기에 하나의 顆粒으로 융합하지 않고 서로 연결된 wtin-tubes로 나타나며, sperm bundle에서는 basal bodies group을 형성한다.

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Positional cloning in mice: a new mutant mouse, Sims (Sexual Immaturity, Megaencephaly, and Seizure)

  • Koo, S.K.;Jin, S.J.;Lee, K.S.;Oh, B.S.
    • 한국동물학회:학술대회논문집
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    • 한국동물학회 1999년도 한국생물과학협회 학술발표대회
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    • pp.31-31
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    • 1999
  • Characterization of mutant mice has been utilized as an animal model for the study of human inherited diseases. In addition to the pathogenesis stduy using the mutant mice, the mice have been used for the identification of the genes causing the phenotypes. Functional cloning and positional cloning are two approaches, depending on the phenotypes of the mutant mice. Though it takes a long time positional cloning has been well used to identify the gene of which function can not be presumed from the mouse phenotype. Recently by the advance of the molecular tools and the human genome project close to 10,000 genetic markers are developed to make the procedure faster. We obtained a new mutant mouse, sims, spontaneously arose and the affected mouse has a mild tremor and seizure was observed. Homozygote in either sex is sterile since uterus growth in female and seminal vesicle in male are not induced for the growth in puberty, implying the abnormal hormonal regulation during puberty. Supporting this, there is no detectable testosterone in the serum of the mutant male and the brain of the mutant is 30% heavier than littermate. To identify the location of the mutated gene, intraspecies cross to CAST/Ei was carried out and the 37 affected mice was analyzed for the linkage. The gene was mapped on chromosome 18, 20 cM from the centromere. More than 500 F2 progenies have been analyzed for the linkage and the locus becomes narrow within 3cM between Egrl and Fgf gene.f gene.

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