• Title/Summary/Keyword: Cortical cell culture

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S-nitrosation Ameliorates Homocysteine-mediated Neurotoxicity in Primary Culture of Bat Cortical Neurons (흰쥐 대뇌피질 신경세포에 미치는 호모시스틴의 신경독성에 대한 S-nitrosation의 역할)

  • Kim, Won-Ki
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.169-175
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    • 1996
  • The reactivity of the sulfhydryl (thiol) group of homocysteine has been associated with an Increased risk of atherosclerosis, thrombosis and stroke. Thiols also react with nitric oxide (NO, an endothelium-derived relaxing factor (EDRF) ), forming S-nitrosothiols that have been reported to have potent vasodilatory and antiplatelet effects and been expected to decrease adverse vascular effects of homocysteine. The present study was aimed to Investigate whether the S-nitrosation of homocysteine modulates the neurotoxic effects of homocysteine. An 18 hour-exposure of cultured rat cortical neurons to homocysteine ( >1 mM) resulted in a significant neuronal cell death. At comparable concentrations ( <10 mM), however, S-nitrosohomocysteine did not induce neuronal cell death. Furthermore, S-nitrosohomocysteirle partially blocked NMDA-mediated neurotoxicity. S-nitrosohomocysteine also decreased NMDA-mediated increases in intracellular calcium concentration. The present data indicate that in brain nitric oxide produced from neuronal and nonneuronal cells can modulate the potential, adverse properties of homocysteine.

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Effects of Spermine on Quisqualate-induced Excitotoxicity in Rat Immature Cortical Neurons (흰쥐 미숙 대뇌피질 신경세포에서 Quisqualate로 유발된 흥분성 세포독성에 대한 spermine의 영향)

  • 조정숙
    • YAKHAK HOEJI
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    • v.43 no.4
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    • pp.535-540
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    • 1999
  • Glutamate (Glu) receptor-mediated excitoxicity has been implicated in many acute and chronic types of neurological disorders. Exposure of mature rat cortical neurons (15-18 days in culture) to the various concentrations of Glu resulted in a marked neuronal death, whereas immature rat cortical neurons (4∼5 days in culture) were resistant to the Glu-induced toxicity. Glu receptor subtype-specific agonists showed differential extent of toxicity in the immature neurons. The neurons treated with NMDA or kainate (KA) did not exhibit damage. However, quisqualate (QA) treatment induced a considerable cell death (36.1%) in immature enurons. The non-NMDA antagonist DNQX did not reduce this response. Interestingly, the QA-induced toxicity was potentiated by spermine in a concentration-dependent manner. Again, the spermine-enhanced damage was not altered by the polyamine antagonist ifenprodil. Taken together, unlike NMDA or KA, QA can induce neurotoxicity in immature rat cortical neurons and the QA-induced toxicity was potentiated by spermine. The lack of antagonizing effects of DNQX and ifenprodil on QA-induced toxicity and the potentiated toxicity by spermine, respectively, implies that both QA receptor and the polyamine site of NMDA receptor may not mediate the neurotoxicity observed in this study, and that a distinct mechanism(s) may be involved in excitotoxicity in immature neurons.

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Influence with Pressure of the Bone Fluid in Inclination of Osteon

  • Yoon, Young-June;Chung, Jae-Pil
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.3 no.4
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    • pp.10-15
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    • 2010
  • Cortical bone is composed of an osteon, which is a subunit of the cortical bone. At the center of the osteon, Haversian is located and it consists of blood vessels and nerves. Osteon is known to be inclined 5 to 15 degrees with respect to the long axis of a cortical bone, but the reason why it is inclined is not clear. Using the poroelastic calculation provides the pore pressure varies at the lacunar-canalicular network from -200KPa to 200KPa. This estimation is close to the result shown in the previous literature and it helps further cell culture experiment for elucidating the bone remodeling process.

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Suppression of Reactive Oxygen Species Production by Water-extracts of Coptidis Rhizoma Enhances Neuronal Survival in a Hypoxic Model of Cultured Rat Cortical Cells. (흰쥐 대뇌세포의 저산소증 모델에서 황련의 활성산소 생성 억제와 신경세포사 억제)

  • Choi, Ju-Li;Shin, Gil-Jo;Lee, Won-Chul;Moon, Il-Soo;Jung, Seung-Hyun
    • Journal of Life Science
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    • v.18 no.3
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    • pp.311-317
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    • 2008
  • Pathophysiological oxidative stress results in neuronal cell death mainly due to the generation reactive oxygen species (ROS). In low oxygen situation such as hypoxia and ischemia, excessive ROS is generated. Coptidis Rhizoma (CR) is a traditional medicine used for the incipient stroke. In this report we show that CR water extracts $(1\;{\mu}g/ml)$ exhibited protective effects of neuronal cell death in a hypoxic model (2% $O_2/5%\;CO_2,\;37^{\circ}C,$ 3 hr) of cultured rat cortical cells. We further show that CR water extracts significantly reduced the intensity of green fluorescence after staining with $H_2DCF-DA$ on one hour and three days after hypoxic shock and in normoxia as well. Our results indicate that CR water extracts prevent neuronal death by suppressing ROS generation.

Primary Cultured Brain Cells as Screening Methods for Natural Products Acting on Glutamatergic Neurons (일차배양 뇌세포를 이용한 글루타메이트성 신경에 작용하는 천연물의 검색방법)

  • 박미정;김소라;문애리;김승희;김영중
    • YAKHAK HOEJI
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    • v.39 no.4
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    • pp.444-449
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    • 1995
  • Primary cultures of rat cortical and chicken embryonic brain cells were employed to establish a reliable screening method for natural products blocldng or enhancing glutamate-induced neurotoxicity. Exposure of primary cultured rat cortical cells or chicken embryonic brain cells to high dose of glutamate resulted in the fragmentation of neutites and consequent neuronal death. The level of cytoplasmic lactate dehydrogenase(LDH), indicator for cell survival in cultures, was significantly reduced at exposure to glutamate. For the practical application of the methods, series of concentrations of plants extracts and positive control were applied prior to the glutamate insult on primary cultures of rat cortical and chicken embryonic, brain cells. Relative LDH level in cells was measured for the estimation of the effect of the test materials on the glutamatergic neurons. The validity of the present screening method for natural products acting on glutamatergic neurons was examined with dextromethorphan, a known glutamatergic antagonist. The treatment of 100 $\mu{M}$ dextromethorphan prevented the reduction of LDH in rat cortical and chicken embryonic brain cells caused by glutamate insult keeping 60% and 90% of LDH level in normal control, respectively. Above results indicate that primary cultures of rat cortical and chicken embryonic brain cells could be proper systems for the screening of potential natural agents acting on glutamatergic, neurons. Between the two types of cultures, primary culture of chicken embryonic brain cells seemed to be a better system for the primary screening, since it is technically easier and economical compared to that of rat cortical cells.

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Prevention of ROS Production and MMP Dissipation by Fructus Schisandrae(FS) in a Hypoxic Model of Cultured Rat Hippocampal Neurons (오미자(五味子)(Fructus Schisandrae)가 흰쥐 해마신경세포의 저산소증 모델에서 세포사에 미치는 영향)

  • Ju, Dae-Hwan;Shin, Gil-Cho;Moon, Il-Soo
    • The Journal of Internal Korean Medicine
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    • v.29 no.4
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    • pp.835-845
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    • 2008
  • Objectives : The purpose of this study was to investigate the effect of FS for the modulation of ROS and MMP in a hypoxic model of cultured rat cortical cells. Methods : For the effect of FS on the viability, FS was added to culture media (neurobasal supplemented with B27) and cell viability was measured by LDH assay. To investigate the effects of FS on ROS generation and MMP preservation, cells grown in FS-containing media were given a hypoxic shock(2% $O_2/5%$ $CO_2$, $37^{\circ}C$, 3 hrs) on DIV 10, stained with $H_2DCF-DA$(10 nM) and JC-1, respectively, and observed by fluorescent microscope. Results : 1. FS has a protective effect of cortical cells in both normoxia and hypoxia. 2. FS reduced the generation of ROS and this reduction was especially significant at 3 days after hypoxia. 3. FS was effective for the maintenance of MMP in hypoxia, and this efficacy was especially significant at 3 days after hypoxia. Conclusions : Taken together, these results indicate that FS attenuates ROS generation and MMP dissipation, which eventually protects from neuronal cell death in hypoxia.

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Neuroprotective Effects of Pinelliae Rhizoma Water-Extract by Suppression of Reactive Oxygen Species and Mitochondrial Membrane Potential Loss in a Hypoxic Model of Cultured Rat Cortical Cells. (배양대뇌신경세포 저산소증모델에서 유해산소생성억제 및 사립체막전위 소실방지에 의한 반하(半夏)의 신경세포사 억제 효능)

  • Kwon, Gun-Rok;Moon, Il-Soo;Lee, Won-Chul
    • Journal of Life Science
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    • v.19 no.5
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    • pp.598-606
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    • 2009
  • Oxidative stress by free radicals is a major cause of neuronal cell death. Excitotoxicity in hypoxia/ischemia causes an increase in reactive oxygen species (ROS) and a loss of mitochondrial membrane potential (MMP), resulting in dysfunction of the mitochondria and cell death. Pinelliae Rhizoma (PR) is a traditional medicine for incipient stroke. We investigated the effects of PR Water-Extract on the modulation of ROS and MMP in a hypoxic model using cultured rat cortical cells. PR Water-Extract was added to the culture medium at various concentrations (0.25${\sim}$5, 5.0 ${\mu}g/ml$) on day in vitro 12(DIV12), given a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 hr), and cell viability was assessed on DIV15 by Lactate Dehydrogenase Assay (LDH assays). PR Water-Extract showed a statistically significant effect on neuroprotection (10${\sim}$15% increase in viability; p<0.01) at 1.0 and 2.5 ${\mu}g/ml$ in normoxia and hypoxia. Measurement of ROS production by $H_2DCF-DA$ stainings showed that PR Water-Extract efficiently reduced the number and intensity of ROS-producing neurons, especially at 1 hr post shock and DIV15. When MMP was measured by JC-1 stainings, PR Water-Extract efficiently maintained high-energy charged mitochondria. These results indicate that PR Water-Extract protects neurons in hypoxia by preventing ROS production and preserving the cellular energy level.

Development of Schizogenous and Lysigenous Aerenchyma in Rice Root

  • Kang, Si-Yong;Wada, Tomikichi;Choi, Kwan-Sam
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.2
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    • pp.77-82
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    • 1998
  • Aerenchyma development in rice (Oryza sativa L.) roots is quite important for adaptation to waterlogged or reduced soil conditions. Anatomical observations were carried out to clarify the development of schizogenous and lysigenous aerenchyma in elongating crown roots of rice. The crown roots of 3rd and 4th phytomer were taken from rice plants of the 8th leaf stage grown by hydroponic culture. The schizogenous intercellular spaces in the cortex of crown root tip were observed using a light microscope with semi ultra-thin sections and the lysigenous aerenchyma in mature tissue of crown root were observed using a cryo scanning electron microscope (cryo-SEM) with freezing fracture method. The schizogenous intercellular spaces in the root tip exist obviously in the middle portion of cortical cell layers close to the root-root cap junction, but not in root cap, stele and outer cell layers of cortex. The air spaces were formed at the junction of four neighbouring cells of inner cortex in the transverse sections, and between longitudinal cell layer connected along the root axis. Although many of those spaces were filled with liquid, some spaces seem to exist as air spaces. The lysigenous aerenchyma in the cortex, which hardly filled with liquid, emerged at 3-4 cm segment from the root tip and increased toward the basal region of root axis. The developing process of lysigenous aerenchyma was primarily separation of a radial row of cells caused by the shrinking and collapsing of cortical cells and then formation of septa along the radial cell rows by the fusion of cell wall with each other. These results suggest that the schizogenous and lysigenous aerenchyma playa role as a passage for the movement of oxygen into the root tip region where oxygen is required for respiration.

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Effects of Water-extract Mixture of Scutellariae baicalensis GEORGI, Acarus gramineus SOLAND and Gastrodia elata BLUME on Cultured Rat Cortical Neurons and Enhancement of Learning and Memory Power (황금(黃芩), 석창포(石菖蒲), 천마(天麻) 물추출액 혼합물이 배양한 흰쥐 대뇌신경세포의 활성과 학습능력 증진 효능)

  • Bae, Chul-Hwan;Jung, Hyun-Jung;Jung, Seung-Hyun;Moon, Il-Soo;Lee, Won-Chul;Sin, Gil-Jo
    • Journal of Life Science
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    • v.19 no.6
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    • pp.756-764
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    • 2009
  • Scutellariae baicalensis GEORGt Acorus gramineus SOLAND and Gastrodia elata BLUME are traditional medicines used in the treatment of incipient stoke. In this study we investigated their effects on various aspects of neuronal differentiation in single or composite forms. Water-extracts of these medicines showed neuroprotective effects on cultured rat cortical neurons in normoxia and hypoxia. To understand the mechanism for neuroprotection we carried out various cell biological assays. They stimulated initial differentiation of neuronal development (transition from stage 1 to 2), and increased the number of spines and the length and number of dendritic processes. These effects were best manifested in the experimental group, which were given a mixture of the three kinds of extracts (p<0.01). To assess improvement of brain functions we carried out Morris water-maze tests for the mice that were fed on these extracts instead of water for 4 weeks. The experimental groups, especially those which were given the mixture of the three kinds of extract, showed significant (p<0.01) enhancement in memory as early as one day after the learning trial. These results indicate that these three kinds of extracts have synergistic effects on neuronal protection and improvement of brain functions.