• Title/Summary/Keyword: Glutamate

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A Glutamate Oxidase-based Biosensor for the Determination of Glutamate (Glutamate Oxidase를 이용한 Glutamate 측정용 Biosensor의 개발)

  • Lee, Young-Chun;Lee, Sang-Hyun
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1075-1081
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    • 1997
  • The objective of this research was to develop a glutamate enzyme sensor for rapid determinations of glutamate in samples. Glutamate oxidase was immobilized onto activated nylon, chitosan and other membranes. The enzymic and nonactin membranes were attached to an ammonia electrode to detect ammonia generated by the reaction between glutamate oxidase and glutamate. The enzyme immobilized on activated nylon membrane was stable for 2 months, and was able to perform about 250 glutamate determinations without losing activities. The enzyme immobilized on chitosan membrane had higher enzyme activity, but was not as much stable as that immobilized on nylon. The glutamate biosensor was able to accurately determine $0.1{\sim}5\;mM$ of glutamate in samples.

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Glutamate-Induced Serotonin Depletion in Fetal Rat Brainstem Cultures (흰쥐태 뇌간의 배양에서 Glutamate에 의한 Serotonin의 고갈)

  • Park, Sang-Wook;Wie, Myung-Bok;Song, Dong-Keun;Kim, Yong-Sik;Kim, Yung-Hi
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.189-193
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    • 1993
  • Exposure of dissociated cultures from fetal rat brainstem to glutamate for upto 6 h decreased cellular contents of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in a concentration- and time-dependent manner. In addition, glutamate induced lactate dehydrogenase leakage. Tetrodotoxin did not block the effects induced by glutamate. MK-801 $(1{\mu}M)$, an N-methyl-D-aspartate (NMDA) channel blocker, but not 6-cyano-2,3-dihydroxy-7-nitro-quinoxazoline $(CNQX;\;3{\mu}M)$, a non-NMDA receptor antagonist, blocked glutamate-induced effects, indicating that these glutamate-induced responses are mediated through NMDA receptors.

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Responsiveness of Dendrites to the Glutamate Applied Focally with Pressure Ejector and Iontophoresis into Hippocampal Slices

  • Kim, Jin-Hyuk;Shin, Hong-Kee;Chang, Hyun-Ju;Kim, Hye-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.6
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    • pp.457-466
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    • 2001
  • Glutamate is the most common excitatory amino acid in the brain. Responsiveness of dendrites to the glutamate greatly varies depending on the application sites. Especially, a point of the maximal response to the glutamate of the dendrite is called as 'hot spot'. In our experiment, the responsiveness of the hot spot to the glutamate was investigated in the CA1 pyramidal neuron of the rat hippocampal slice. CNQX, the antagonist of AMPA receptor, blocked 95% of membrane current to the glutamate focal application $(I_{gl}).$ Train ejection of glutamate on one point of the dendrite increased or decreased the amplitude of $I_{gl}$ with the pattern of train, and the changes were maintained at least for 30 min. In some cases, glutamate train ejection also induced calcium dependent action potentials. To evoke long-term change of synaptic plasticity, we adopted ${\theta}-burst$ in the glutamate train ejection. The ${\theta}-burst$ decreased the amplitude of glutamate response by 60%. However, after ${\theta}-burst$ glutamate train ejection, the calcium dependent action potential appeared. These results indicated that the focal application of glutamate on the neuronal dendrite induced response similar to the synaptic transmission and the trains of glutamate ejection modulated the change of AMPA receptor.

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Effects of betaine on the glutamate-induced neurotoxicity in primary cultured chicken brain cells

  • 김영중
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.46-46
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    • 1993
  • The neuroprotective effect of betaine, one of the , components of Lycii Fructus, on glutamate-induced neurotoxicity in primary cultured chicken brain cells were examined. Betaine was found to attenuate glutamate-induced neurotoxicity at the concentration of 5-10 mM in both morphological and chemical aspects. The pretreament of chicken brain cells with 5-10 mM betaine for 2 hr at the 12th day of culture before the 40 min-exposure to 500${\mu}$M glutamate significantly increased the survival rate of nerve cells in chicken brain. Betaine could also raise the decreased LDH-level due to the neurotoxicity induced with 100${\mu}$M glutamate in chicken braill cells. LDH value was decreased to 63% of control level in chicken brain cells at the time of 48 hr after the exposure to glutamate. However, the pretreament of chicken brain cells with 5 mM betaine for 2 hr before the exposure to glutamate could prevent the decrease of LDH-level in brain cells showing 90% of control level. Nevertheless, tile remarkable neuroprotective effect of betaine on the glutamate-inducer in neurotoxicity in cultured chicken brain cells could not be observe when betaine was simultaneously administered with glutamate.

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Effects of Oxygen Free Radicals on Extracellular Glutamate Accumulation in Cultured Cells

  • Shin, Chang-Sik;Oh, Seikwan;Lee, Myung-Koo;Lee, Myung-Koo;Kim, Hack-Seang
    • Archives of Pharmacal Research
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    • v.19 no.2
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    • pp.132-136
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    • 1996
  • Exogenously applied oxygen free radical generating agent, pyrogallol, highly elevated extracellular glutamate accumulation and augmented N-methyl-D-aspartate (NMDA)-induced glutamate accumulation in cerebellar granule neuronal cells, but did not in astrocytes. Superoxide dismutase remarkably decreased the pyrogallol-induced glutamate accumulation, but either NMDA or kainate antagonists did not. In addition, pyrogallol did not affect the NMDAinduced intracellular calcium elevation. Pyrogallol partially blocked glutamate uptake into astrocytes. These results suggest that oxygen free radicals elevate extracellular glutamate accumulation by stimulating the release of glutamate as well as blocking the glutamate uptake.

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Effects of Chronic Lead Exposure on Glutamate Release and Uptake in Cerebellar Cells of Rat Pups

  • Yi, Eun-Young;Lim, Dong-Koo
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.113-119
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    • 1998
  • Changes in the release and uptake of glutamate in cerebellar granule and glial cells of offspring of lead-exposed mothers were determined. In cultured cerebellar granule cells exposed to lead for 5 days, glutamate release was less influenced upon N-methyl-D-aspartate (NMDA) stimulation than that in the control. Although the NMDA-stimulated release of glutamate in cerebellar granule cells prepared from lead-exposed first generation pups was not different from that of the control group, the S-nitroso-N-acetylpenicillamine (SNAP)-stimulated release of glutamate in cerebellar granule cells obtained from lead-treated pups was less elevated than that in the control. Furthermore, in cerebellar granule cells obtained from lead-exposed second generations pups, glutamate release did not respond to both NMDA and SNAP stimulation. In cerebellar glial cells exposed to lead, the basal glutamate uptake was not changed. However, the L-trans-pyrollidine-2,4-dicarboxylic acid (PDC)-blocking effects was significantly reduced. In glial cells obtained from lead-exposed pups, the glutamate uptake was also less blocked by PDC than that in the control. Further decreases in PDC-blocking effects were observed in cerebellar glial cells obtained from lead-treated second generation pups compared to those from the control group. These results indicate that lead exposure induces the changes in the sensitivities of the glutamate release and uptake transporter. In addition, these results suggest that lead exposure might affect the intracellular signalling pathway and transmission in glutamatergic nervous system.

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Effects of Gwibitang on Glutamate-induced Apoptosis in C6 Glial Cells (귀비탕이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향)

  • 강익현;이인;한상혁;문병순
    • The Journal of Korean Medicine
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    • v.22 no.4
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    • pp.45-57
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    • 2001
  • Objectives : The water extract of Gwibitang (GBT) has been traditionally used for treatment of psychologic disease and brain damage in Oriental Medicine, This study was designed to investigate the effect of GBT on the glutamate-induced toxicity of rat C6 glial cells. Methods : The cultured cells were pretreated with GBT and exposed to glutamate, The cell damage was assessed by using MTT assay and Hoechst, IC-l staining, Results : GBT had protective effects in glutamate-induced cytotoxicity, which was revealed as apoptosis characterized by chromatic condensation and the loss of mitochondrial membrane potential in C6 glial cells. However, GBT and glutamate had no effect in the activation of caspase family cysteine proteases including caspase-3, -8 and -9 proteasesin C6 glial ce]]s, GBT significantly recovered the depletion of GSH and inhibited the generation of $H_2O_2$ by glutamate in C6 glial cells. In addition, both GBT and antioxidants such as GSH and NAC protected the glutamate-induced cytotoxicity in C6 glial cells, indicating that GBT possibly has antioxidative effect. Moreover, GBT also inhibited the glutamate-induced degradation of $IkB{\alpha}$ in C6 glial cells, This result suggest that GBT has some inhibitory effects on the transcriptional activation of $NF-_{k}B$. Conclusions : GBT has protective effects in glutamate-induced cytotoxicity via an antioxidative mechanism.

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Inhibitory Effects of Ginsenosides on Glutamate-Induced Swelling of Cultured Astrocytes

  • Seong, Yeon-Hee;Koh, Sang-Bum;Kim, Hack-Seang
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.138-142
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    • 2000
  • Effects of ginsenosides (Rb$_1$, Rb$_2$, Rc, Re, Rg$_1$, Rf) on L-glutamate (glutamate)-induced swelling of cultured astrocytes from rat brain cerebral cortex were studied. Following the exposure to 0.5mM glutamate for 1 hr, the intracellular water space (as measured by [$^3$H]O-methyl-D-glucose uptake) of astrocytes increased by about two-fold. Simultaneous addition of ginsenosides Rb$_2$ and Rc with glutamate reduced the astrocytic swelling in a dose-dependent manner. These ginsenosides at 0.5 mg/ml did not affect the viability of astrocytes for up to 24 hr which was determined by a colorimetric assay (MTT assay) for cellular growth and survival. These ginsenosides at 0.3 mg/ml inhibited the increase of intracellular Ca$\^$2+/ concentration ([Ca$\^$2+/]$\_$i/) induced by glutamate. These data suggest ginsenosides Rb$_2$ and Rc prevent the cell swelling of astrocytes induced by glutamate, maybe via inhibition of Ca$\^$2+/ influx.

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Effect of Yukgunja-tang on Glutamate-induced Apoptosis in C6 Glial Cells (육군자탕(六君子湯)이 Glutamate에 의한 C6 신경교세포의 Apoptosis에 미치는 영향)

  • Jang, Won-Seok;Shin, Yong-Jeen;Ko, Seok-Jae;Ha, Ye-Jin;Kwon, Young-Mi;Shin, Sun-Ho
    • The Journal of Internal Korean Medicine
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    • v.31 no.3
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    • pp.586-599
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    • 2010
  • Objective : The water extract of Yukgunja-tang(YGJT) has been traditionally used in treatment of qi deficiency and phlegm in Oriental medicine. However, little is known about the mechanism by which YGJT protects neuronal cells from injury damages. Therefore, this study was designed to evaluate the protective effects of YGJT on C6 glial cells by glutamate-induced cell death. Methods : The present study describes glutamate, which is known as an excitatory neurotransmitter, related with oxidative damages, and YGJT, which shows protective effects against glutamate-induced C6 glial cell death. One of the main mediators of glutamate-induced cytotoxicity was known on the generation of reactive oxygen species(ROS) via activation of NADPH oxidase (NOX). The protective effects of antioxidant(NAC) and NOX inhibitor(apocynin) on the glutamate-induced C6 glial cells were determined by a MTT reduction assay. Result : YGJT inhibited glutamate-induced ROS generation via inhibition of NOX expression on glutamate-stimulated C6 glial cells. Furthermore, YGJT attenuated glutamate-induced caspase activation. These results suggest that YGJT could be a new potential candidate against glutamate-induced oxidative stress and cell death. Conclusion : These findings indicate that in C6 glial cells, ROS plays an important role of glutamate-induced cell death and that YGJT may prevent cell death from glutamate-induced cell death by inhibiting the ROS generation.

Quercetin ameliorates glutamate toxicity-induced neuronal cell death by controlling calcium-binding protein parvalbumin

  • Kang, Ju-Bin;Park, Dong-Ju;Shah, Murad-Ali;Koh, Phil-Ok
    • Journal of Veterinary Science
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    • v.23 no.2
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    • pp.26.1-26.12
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    • 2022
  • Background: Glutamate is the main excitatory neurotransmitter. Excessive glutamate causes excitatory toxicity and increases intracellular calcium, leading to neuronal death. Parvalbumin is a calcium-binding protein that regulates calcium homeostasis. Quercetin is a polyphenol found in plant and has neuroprotective effects against neurodegenerative diseases. Objectives: We investigated whether quercetin regulates apoptosis by modulating parvalbumin expression in glutamate induced neuronal damage. Methods: Glutamate was treated in hippocampal-derived cell line, and quercetin or vehicle was treated 1 h before glutamate exposure. Cells were collected for experimental procedure 24 h after glutamate treatment and intracellular calcium concentration and parvalbumin expression were examined. Parvalbumin small interfering RNA (siRNA) transfection was performed to detect the relation between parvalbumin and apoptosis. Results: Glutamate reduced cell viability and increased intracellular calcium concentration, while quercetin preserved calcium concentration and neuronal damage. Moreover, glutamate reduced parvalbumin expression and quercetin alleviated this reduction. Glutamate increased caspase-3 expression, and quercetin attenuated this increase in both parvalbumin siRNA transfected and non-transfected cells. The alleviative effect of quercetin was statistically significant in non-transfected cells. Moreover, glutamate decreased bcl-2 and increased bax expressions, while quercetin alleviated these changes. The alleviative effect of quercetin in bcl-2 family protein expression was more remarkable in non-transfected cells. Conclusions: These results demonstrate that parvalbumin contributes to the maintainace of intracellular calcium concentration and the prevention of apoptosis, and quercetin modulates parvalbumin expression in glutamate-exposed cells. Thus, these findings suggest that quercetin performs neuroprotective function against glutamate toxicity by regulating parvalbumin expression.