• Title/Summary/Keyword: mGluR

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Electrophysiological Characterization of AMPA and NMDA Receptors in Rat Dorsal Striatum

  • Jeun, Seung-Hyun;Cho, Hyeong-Seok;Kim, Ki-Jung;Li, Qing-Zhong;Sung, Ki-Wug
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.3
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    • pp.209-214
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    • 2009
  • The striatum receives glutamatergic afferents from the cortex and thalamus, and these synaptic transmissions are mediated by ${\alpha}$-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and N-methyl D-aspartate (NMDA) receptors. The purpose of this study was to characterize glutamate receptors by analyzing NMDA/AMPA ratio and rectification of AMPA and NMDA excitatory postsynaptic currents (EPSCs) using a whole-cell voltage-clamp method in the dorsal striatum. Receptor antagonists were used to isolate receptor or subunit specific EPSC, such as (DL)-2-amino-5-phosphonovaleric acid (APV), an NMDA receptor antagonist, ifenprodil, an NR2B antagonist, CNQX, an AMPA receptor antagonist and IEM-1460, a GluR2-lacking AMPA receptor blocker. AMPA and NMDA EPSCs were recorded at - 70 and + 40 mV, respectively. Rectification index was calculated by current ratio of EPSCs between + 50 and - 50 mV. NMDA/AMPA ratio was 0.20${\pm}$0.05, AMPA receptor ratio of GluR2-lacking/GluR2-containing subunit was 0.26${\pm}$0.05 and NMDA receptor ratio of NR2B/NR2A subunit was 0.32${\pm}$0.03. The rectification index (control 2.39${\pm}$0.27) was decreased in the presence of both APV and combination of APV and IEM-1460 (1.02${\pm}$0.11 and 0.93${\pm}$0.09, respectively). These results suggest that the major components of the striatal glutamate receptors are GluR2-containing AMPA receptors and NR2A-containing NMDA receptors. Our results may provide useful information for corticostriatal synaptic transmission and plasticity studies.

Distribution of Glutamate Receptors in the Retina of the Greater Horseshoe Bat (Rhinolophus ferrumequinum) (한국관박쥐 망막에서 글루타메이트 수용체의 분포 양상)

  • Kwon, Oh-Ju;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.3
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    • pp.413-418
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    • 2014
  • Purpose: The objective of this study was analyzing the distribution of the excitatory neurotransmitter glutamate receptor to investigate the function in the retina of the greater horseshoe bat. Methods: After retinal tissues of adult greater horseshoe bat were cut into $40{\mu}m$ vertical sections, standard immuno-cytochemical techniques was applied for analysis. Immunofluorescence images were obtained using the Bio-Rad MRC 1024 laser scanning confocal microscope. Results: AMPA (GluR1-4), Kainate (GluR5-7, KA1-2) and NMDA (1, 2A, 2B) mainly distributed in the inner plexiform layer and outer plexiform layer. KA1 receptors have existed not only plexiform layer but also ganglion cell layer. Conclusions: The greater horseshoe bat has same neuron and neurotransmitter to mammalian retina. These findings suggest that bat has a functional retina for visual analysis.

Structure and Photoreaction of Photoactive Yellow Protein

  • Imamoto, Yasushi;Harigai, Miki;Shimizu, Nobutaka;Kamikubo, Hironari;Yamazaki, Yoichi;Kataoka, Mikio
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.126-129
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    • 2002
  • The chromophore/protein interactions in the photocycle intermediates of photoactive yel- low protein (PYP) were probed by site-directed mutagenesis. The absorption spectra of L- intermediates produced from E46Q, T50V, and R52Q mutants were calculated using the absorption spectra of dark states and difference absorption spectra between L-intermediates and dark states, and compared with that of PYP$\_$L/. The absorption spectrum of R52Q$\_$L/ agreed with that of PYP$\_$L/, but those of E46Q$\_$L/ and T50V$\_$L/ were red-shifted. The effect of these mutations on the absorption spectrum for L-intermediate was comparable to that for the dark state, suggesting that the interaction around the phe-nolic oxygen of the chromophore is conserved in PYP$\_$L/ unlike the crystal structure. On the other hand, we have reported that the absorption spectra of Y 42F$\_$M/, T50V $\_$M/, and R52Q$\_$M/ agreed with that of PYP$\_$M/, but that of E46Q$\_$M/ was red-shifted, suggesting that the hydrogen bond of the chromophore with Glu46 is conserved but that with Tyr42 is broken in PYP$\_$M/. These results suggest that the chromophore inter-acts with Glu46 throughout the photocycle, but never directly interacts with Arg52. This model con- flicts with some of the structural model of PYP intermediates proposed based on the high-resolution X -ray crystallography.

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Development of Saccharomyces cerevisiae Reductase YOL151W Mutants Suitable for Chiral Alcohol Synthesis Using an NADH Cofactor Regeneration System

  • Yoon, Shin Ah;Jung, Jihye;Park, Seongsoon;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.218-224
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    • 2013
  • The aldo-keto reductases catalyze reduction reactions using various aliphatic and aromatic aldehydes/ketones. Most reductases require NADPH exclusively as their cofactors. However, NADPH is much more expensive and unstable than NADH. In this study, we attempted to change the five amino acid residues that interact with the 2'-phosphate group of the adenosine ribose of NADPH. These residues were selected based on a docking model of the YOL151W reductase and were substituted with other amino acids to develop NADH-utilizing enzymes. Ten mutants were constructed by site-directed mutagenesis and expressed in Escherichia coli. Among them, four mutants showed higher reductase activities than wild-type when using the NADH cofactor. Analysis of the kinetic parameters for the wild type and mutants indicated that the $k_{cat}/K_{m}$ value of the Asn9Glu mutant toward NADH increased 3-fold. A docking model was used to show that the carboxyl group of Glu 9 of the mutant formed an additional hydrogen bond with the 2'-hydroxyl group of adenosine ribose. The Asn9Glu mutant was able to produce (R)-ethyl-4-chloro-3-hydroxyl butanoate rapidly when using the NADH regeneration system.

Anti-apoptotic and Neuroprotective Effects of Acupuncture at $LR_3$ on Focal Brain Ischemic Injury Induced by Intraluminal Filament Insertion in Rats (다종(多種)의 태충(太衝)($LR_3$) 침척요법(鍼刺療法)이 Intraluminal Filament 삽입술로 유발된 백서(白鼠)의 허혈성 국소 뇌손상에 미치는 영향)

  • Yim, Hyun-Jin;Cho, Myung-Rae;Youn, Dae-Hwan;Na, Chang-Su;Ryu, Chung-Ryeol
    • Journal of Acupuncture Research
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    • v.24 no.2
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    • pp.125-140
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    • 2007
  • Objectives: This study was performed to investigate the effects of acupuncture therapy(AT, AT-9), electro-acupuncture therapy(EAT) and low level laser acupuncture therapy(LAT) at LRJ on the focal ischemia-induced by intraluminal filament insertion in rats. Methods : In the present syudy, the focal ischemia was induced by Intraluminal filament insertion into left middle cerebral artery. The subjects were divided into five groups after focal brain ischemia. (n=15, in each group) : Control with no treatment, AT with acupuncture at $LR_3$, AT-9 with acupuncture at $LR_3$ and rotating 9 times in a clockwise direction, EAT with electro-acupuncture at $LR_3$ and LAT with invasive laser acupuncture at $LR_3$. Anti-apoptotic and neuroprotective effects of acupuncture were observed by mGluR5 mRNA, Bax mRNA, Bcl-2 mRNA, Cytochrome C protein, Cresyl violet-stain and Choline acetyltransferase (ChAT)-stain in the hippocampus. Results: 1. In LAT, mGluR5, Cresyl violet-stain and ChAT-stain were increased. 2. In LAT, Cytochrome C protein was decreased. 3. In AT-9, Bax, Cytochrome C protein and the Bax/Bcl-2 ratio were decreased. 4. In AT-9, Bcl-2, Cresyl violet-stain and ChAT-stain were increased. 5. In EAT, Bcl-2 and Cresyl violet-stain were increased. Conclusions: These results suggests that LAT and AT-9 show anti-apoptotic and neuro-protective effects and that LAT and AT-9 may be useful for managing stroke by focal brain ischemia.

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Effect of Various β-1,3-glucan Supplements on the Performance, Blood Parameter, Small Intestinal Microflora and Immune Response in Laying Hens (β-Glucan 제제들이 산란계의 생산성, 혈액 성상과 소장내 미생물 균총 및 면역 체계에 미치는 영향)

  • Park, K.W.;Rhee, A.R.;Lee, I.Y.;Kim, M.K.;Paik, I.K.
    • Korean Journal of Poultry Science
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    • v.35 no.2
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    • pp.183-190
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    • 2008
  • This study was conducted to investigate the effect of feeding diets supplemented with ${\beta}-glucan$ products on the performance, small intestinal microflora and immune response in laying hens. The ${\beta}-glucan$ products used in the experiment were $BetaPolo^{(R)}$ ; soluble ${\beta}-glucan$ of microbial cell wall origin, $HiGlu^{(R)}$ ; microbial cell wall origin, $OGlu^{(R)}$ ; oat origin, $BGlu^{(R)}$ ; barley origin. A total of 720 Hy-Line Brown laying hens of 40wks old were divided into 5 dietary treatments : T1 ; Control( C), T2 ; $BetaPolo^{(R)}$, T3 ; $HiGlu^{(R)}$, T4 ; $OGlu^{(R)}$, T5 ; $BGlu^{(R)}$. Each treatment was replicated 4 times with 36 birds/replicate housed in 2 bird cages, and arranged according to completely randomized block design. Feeding trial lasted 40ds under 16 h lighting regimens. There were significant differences among treatments in hen-house egg production feed intake and feed conversion. HiGlu treatment was significantly higher than OGlu treatments in hen-house egg production. ${\beta}-glucan$ supplemented treatments were lower than the control in feed intake and feed conversion ratio. All ${\beta}-glucan$ supplemented treatments were significantly higher than the control in eggshell strength. Eggshell color and Haugh unit tended to be lower in the supplemented group than the control. IgY concentration was not significantly affected by treatments. At $5^{th}$ week of experiment, however, IgY concentration tended to increase in the supplemented groups. Among the leucocytes parameters, WBC, heterophil, lymphocytes, monocyte and eosinophil concentration were lower in the supplemented groups than those of the control. Among erythrocytes, HCT(hematocrit) and MCV(mean corpuscular volume) were significantly affected by treatment. MCV of supplemented groups were higher than that of the control. Immunoglobulin concentrations in the birds were not significantly different among treatments. However, IgA concentration tended to be low in the supplemented groups than the control. The cfu of small intestinal microflora were not significantly different among treatments, but that of Cl. perfringens tended to be lower than the control. The result of this experiment indicateted that feeding ${\beta}-glucan$ to laying hens improve feed conversion ratio and eggshell strength. Also intestinal microflora and immune responses are modified.

Effects of acupuncture at the $LU8{\cdot}K17$ on Anti-apoptotic cell death and neuroprotection in Rat hippocampus following focal brain ischemic injury induced by Intraluminal Filament insertion in Rats (직자(直刺), 영수(迎隨) 및 염전수기법(捻轉手技法)에 따라 시행한 경거(經渠)${\cdot}$복류(復溜) 침자(鍼刺)가 중대뇌동맥(中大腦動脈) 폐새(閉塞)에 의하여 유발(誘發)된 국소(局所) 뇌허혈(腦虛血) 백서(白鼠) hippocampus의 항세포자멸사(抗細胞自滅死) 및 복경보호(福經保護)에 미치는 영향(影響))

  • Youn, Dae-Hwan;Byun, Jeng-Yun;Choi, Chan-Hun;Baek, Jin-Ung;Jeong, Ji-Yeon;Jung, Yeon-Jin;Na, Chang-Su
    • Korean Journal of Acupuncture
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    • v.24 no.3
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    • pp.205-221
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    • 2007
  • Objectives : Aims of this study is to investigate the effects of $LU8{\cdot}KI7$ in rat induced by experimental focal ischemia. Materials and methods : The focal ischemia was induced by intraluminal filament insertion into middle cerebral artery. The groups divided into 6groups, control; no therapy group after ischemia-induced, AT1; acupuncture therapy group at $LU8{\cdot}KI7$ after ischemia-induced, AT2; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally and the needle was twirled and rotated forward with the thumb of the right hand 9times, AT3; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally and the needle was twirled and rotated forward with the forefinger of the right hand 9times, AT4; acupuncture therapy at$LU8{\cdot}KI7$ bilaterally and the needle was inserted to the direction following the flowing route of the meridian(digital direction), AT5; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally, the needle was inserted to the direction following the flowing route of the meridian(digital direction) and the needle was twirled and rotated forward with the thumb of the right hand 9times. Acupuncture therapy was carried out 7times during 2weeks after focal ischemia-induced. The anti-apoptotic and neuroprotective effects of acupuncture are observed by Bax, Bcl-2, mGluR5, cytochrome c, Cresyl violet and ChAT-stain. Results : The intensity of Bax was decreased in AC1, AC4, AC5 group, was increased in AC2, AC3 group. The intensity of Bcl-2 was increased in AC2, AC3, AC4, AC5 group. The intensity of mGluR5 was decreased in AC1 group, was increased in AC4, AC5 group. The intensity of Cytochrome c was increased in ACI, AC2 group, was decreased in AC4, AC5 group. The density of neurons stained by Cresyl violet was increased in all group without control group. The density of ChAT was increased in AC2, ACS group. Conclusions : Our study suggests that AC5 group show anti-apoptotic and neuroprotective effects on cholinergic neuron in focal cerebral ischemia of the stroke in rats.

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Cloning and Characterization of Zebrafish Microsomal Epoxide Hydrolase Based on Bioinformatics (생물정보학을 이용한 Zebrafish Microsomal Epoxide Hydrolase 클로닝 및 특성연구)

  • Lee Eun-Yeol;Kim Hee-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.129-135
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    • 2006
  • A gene encoding for a putative microsomal epoxide hydrolase (mEH) of a zebrafish, Danio rerio, was cloned and characterized. The putative mEH protein of D. rerio exhibited sequence similarity with mammalian mEH and some other bacterial EHs. A structural model for the putative mEH was constructed using homology modeling based on the crystallographic templates, 1 qo7 and 1 ehy. The catalytic triad consisting of $Asp^{233}$, $Glu^{413}$, and $His^{440}$ was identified, and the characteristic features such as two tyrosine residues and oxyanion hole were found to be highly conserved. Based on bioinformatic analysis together with EH activity assay, the putative protein was annotated as mEH of D. rerio. Enantiopure styrene oxide with enantiopurity of 99%ee and yield of 33.5% was obtained from racemic styrene oxide by the enantioselective hydrolysis activity of recombinant mEH of D. rerio for 45 min.

Inhibitory and Excitatory Postsynaptic Currents of Medial Vestibular Nucleus Neurons of Rats

  • Chun, Sang-Woo;Choi, Jeong-Hee;Park, Byung-Rim
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.2
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    • pp.59-63
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    • 2003
  • The medial vestibular nucleus (MVN) neurons are controlled by excitatory synaptic transmission from the vestibular afferent and commissural projections, and by inhibitory transmission from interneurons. Spontaneous synaptic currents of MVN neurons were studied using whole cell patch clamp recording in slices prepared from 13- to 17-day-old rats. The spontaneous inhibitory postsynaptic currents (sIPSCs) were significantly reduced by the $GABA_A$ antagonist bicuculline ($20{\mu}M$), but were not affected by the glycine antagonist strychnine ($1{\mu}M$). The frequency, amplitude, and decay time constant of sIPSCs were $4.3{\pm}0.9$ Hz, $18.1{\pm}2.0$ pA, and $8.9{\pm}0.4$ ms, respectively. Spontaneous excitatory postsynaptic currents (sEPSCs) were mediated by non-NMDA and NMDA receptors. The specific AMPA receptor antagonist GYKI-52466 ($50{\mu}M$) completely blocked the non-NMDA mediated sEPSCs, indicating that they are mediated by an AMPA-preferring receptor. The AMPA mediated sEPSCs were characterized by low frequency ($1.5{\pm}0.4$ Hz), small amplitude ($13.9{\pm}1.9$ pA), and rapid decay kinetics ($2.8{\pm}0.2$ ms). The majority (15/21) displayed linear I-V relationships, suggesting the presence of GluR2-containing AMPA receptors. Only 35% of recorded MVN neurons showed NMDA mediated currents, which were characterized by small amplitude and low frequency. These results suggest that the MVN neurons receive excitatory inputs mediated by AMPA, but not kainate, and NMDA receptors, and inhibitory transmission mediated by $GABA_A$ receptors in neonatal rats.