• Archives of Pharmacal Research
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• 제21권2호
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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.

• The Korean Journal of Physiology and Pharmacology
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• 제22권2호
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• pp.155-162
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• 2018

3-(2-Carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), a competitive N-methyl-D-aspartate (NMDA) receptor antagonist, produces rapid antidepressant-like effects in animal models of depression. However, the molecular mechanisms underlying these behavioral actions remain unknown. Here, we demonstrate that CPP rapidly stimulates histone deacetylase (HDAC) 5 phosphorylation and nuclear export in rat hippocampal neurons. These effects are accompanied by calcium/calmodulin kinase II (CaMKII) and protein kinase D (PKD) phosphorylation. Behavioral experiments revealed that viral-mediated hippocampal knockdown of HDAC5 blocked the antidepressant effects of CPP in stressed animals. Taken together, our results imply that CPP acts via HDAC5 and suggest that HDAC5 is a common regulator contributing to the antidepressant actions of NMDA receptor antagonists such as CPP.

• Applied Microscopy
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• 제28권4호
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• pp.591-601
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• 1998

The neurotoxic effects of radiation have been studied in NSC-34 hybrid cells derived from embryonic mouse spinal cord cells. NSC-34 cells irradiated at 25Gy were decreased the cell viability in a time and dose dependent manner. The decrease in cell viability induced by the irradiation was blocked by catalase. Antagonists of the N-methyl-D-aspartate (NMDA) receptor, including D-2-amino-5-phosphonovaleric acid (APV) and chlorokynurenic acid (CKA), similarly blocked radiational induced in cell viability. We performed morphological analysis of light and electron microscope. NSC-34 cells irradiated at 25Gy were decreased the cell density and increased lysosomes and vacuoles in the cytoplasm. Especially chromatin modification was observed. These results indicated that radiation was involved in the oxidant-initiated neurotoxicity and the compounds catalase, APV and CKA were shown to be neuroprotective against radiation.

• 동의생리병리학회지
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• 제17권4호
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• pp.996-1001
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• 2003

It is well known that oxidative stress of reactive oxygen species(ROS) may be a causative factor in the pathogenesis of bone disorder. The purpose of this study was to evaluate the cytotoxicity of oxidative stress. Cell viability by MTS assay or INT assay, activity of glutathione peroxidase(GPx), lipid peroxidation(LPO) activity and cell viablity. And also protctive effect of glutamate receptors against ROS-induced osteotoxicity was examined by protein synthesis, alkaline phosphatase (ALP) activity and lactate dehydrogenase (LDH) activity in cultured rat osteoblasts and osteoclasts. XO/HX decreased cell viability and GPx activity, protein synthesis and ALP activity, but increased LPO activity and LDH activity. In the protective effect, N-methyl-D-aspartate (NMDA) receptor antagonists or AMPA/kainate receptor antagonists such as D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), NMDA receptor antagonists but AMPA/kainate receptor antagonists showed protective effect on xanthine oxidase (XO) and hypoxanthine (HX) in these cultures by the increse of protein synthesis, ALP activity.

• Bulletin of the Korean Chemical Society
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• 제18권9호
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• pp.939-945
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• 1997

Several types of 4-substituted-quinoline-2-carboxylic acid derivatives possessing different substituents at C4-position such as sulfonyl, phosphonyl, carbonyl groups, or a flexible alkyl chain have been synthesized and evaluated for their in vitro antagonistic activity at the glycine site on the N-methyl-D-aspartate (NMDA) receptor. Of them, 5,7-dichloro-4-(tolylsulfonylamino)-quinoline-2-carboxylic acid 9 was found to have the best in vitro binding affinity with IC50 of 0.57 μM. On the other hand, in compounds 21 and 22 the introduction of flexible alkyl chains on C4 of the quinoline mother nuclei caused a significant decrease of the in vitro binding affinity. In addition, replacement of polar carboxylic acid group on C2 by neutral bioisosteres in compounds 23a-d also seems to be disadvantageous to in vitro activity. In the structure-activity relationship (SAR) study of the 4-substituted quinoline-2-carboxylic acid acid derivatives, it was realized that the substitution pattern on C4 significantly influences on the binding affinity for the glycine site of NMDA receptor and the binding affinity might be increased by the introduction of a suitable electron rich substituent at C4 which has the ability of H-bonding donor.

• Investigative Magnetic Resonance Imaging
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• 제12권1호
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• pp.8-19
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• 2008

목적 : 본 연구에서는 인체 뇌 대사물질에 대하여 2D MR 기술인 correlation spectroscopy (COSY) 와 nuclear Overhauser effect/enhancement spectroscopy (NOESY)를 직접 적용하고, 데이터를 획득하여 인체 뇌대사물질들간의 스칼라 짝지움 (coupling)과 쌍극자 (dipolar) 상호작용- NOE에 대한 분석을 통하여 결합연결관계 및 공간연결관계에 대한 정보를 획득하고자 하였다. 대상 및 방법 : 모든 2-D (dimensional) MR 실험 (즉, COSY와 NOESY)은 z축 능동차폐 펄스경사자장, 삼중공명 동결탐침기가 장착된 Bruker Avance 500 (11.8 T) 장비에서 298 K에 수행되었다. MRS상에서 뇌 대사물질과 유사하도록만든 희석액을 만들었고, 최종 MRS 샘플은 10% $D_2O$를 이용하였다. 2-D 스펙트라는 2048 복합 (complex) 데이터 포인트로서 총 320개 의 free induction decay (FID)를 평균화 (averaging)하였고, $H_2O$에서 얻어진 스펙트라는 8012 Hz 였다. 반복지연 (repetition delay) 시간은 2초, 각각의 FID는 4개의 평균화를 선택하였다. 얻어진 2D-COSY, 2D-NOESY 데이터는 Top Spin 2.0 소프트웨어에서 후 처리기법 (post-processing)에 의해 분석되었다. 분석 대상 대사물질은 N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), glutamine (Gln), glutamate (Glu), myo-inositol (Ins), lactate (Lac)로서 총 7 가지 화학물로서 주요 목표 피크로 정했다. 결과 : 인체 뇌 대사물질에 대한 대칭형태의 2D-COSY와 2D-NOESY 스펙트럼을 획득하였고, COSY 스펙트럼상에서는 오직 1.0-4.5 ppm 사이에서만 교차피크들이 생성된 반면 NOESY 스펙트럼상에서는 1.0-4.5 ppm 외에도 7.9 ppm에서 공명 교차피크를 발견할 수 있었다. COSY 스펙트럼을 통하여 lactate에서 메틸 프로톤과 CH 프로톤의 COSY 크로스피크가 발견되었고, NAA에서 메틸렌 프로톤들간과 메틸렌 프로톤과 NH프로톤의 크로스피크가 발견되었고, Ins에서 CH 프로톤 들간의 크로스피크가 발견되었다. NOESY 스펙트럼을 통하여 NAA 분자내 NH 프로톤과 메틸 (-CH3) 프로톤과의 NOESY 크로스피크가 발견되었고, lactate에서 메틸 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Cr에서 메틸 프로톤과 메틸렌 프로톤과의 크로스피크가 발견되었고, Glu에서 메틸렌 프로톤 들간과 또한 메틸렌 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Gln에서 메틸렌 프로톤과 CH 프로톤과의 크로스피크가 발견되었고, Ins에서 CH 프로톤 들간의 크로스피크가 발견되었다. 결론 : 본 연구에서는 in vitro 상태의 인체 뇌 대사물질에 2D-COSY와 2D-NOESY 기술을 직접 적용하고, 결합연결관계 및 공간연결관계에 대한 정보를 성공적으로 획득하여 분석하여 보았다. 본 연구 결과물은 향후 인체 내 in vivo 2D-COSY를 이용한 뇌 대사물질 연구에 매우 유용하리라 사료된다.

• Archives of Pharmacal Research
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• 제22권1호
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• pp.48-54
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• 1999

These studies were designed to examine the differential effect of nitric oxide (NO) and cGMP on glutamate neurotransmission. In primary cultures of rat cerebellar granule cells, the glutamate receptor agonist N-methyl-D-aspartate (NMDA) stimulates the elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), the release of glutamate, the synthesis of NO and an increase of cGMP. Although NO has been shown to stimulate guanylyl cyclase, it is unclear yet whether NO alters the NMDA-induced glutamate release and ${[Ca^{2+}]}_i$ elevation. We showed that the NO synthase inhibitor, NG-monomethyl-L-arginine (NMMA), partially prevented the NMDA-induced release of glutamate and elevation of ${[Ca^{2+}]}_i$ and completely blocked the elevation of cGMP. These effects of NO on glutamate release and [Ca2+]i elevation were unlikely to be secondary to cGMP as the cGMP analogue, dibutyryl cGMP (dBcGMP), did not suppress the effects of NMDA. Rather, dBcGMP slightly augmented the NMDA-induced elevation of ${[Ca^{2+}]}_i$ with no change in the basal level of glutamate or ${[Ca^{2+}]}_i$. The extracellular NO scavenger hydroxocobalamine prevented the NMDA-induced release of glutamate providing indirect evidence that the effect of NO may act on the NMDA receptor. These results suggest that low concentration of NO has a role in maintaining the NMDA receptor activation in a cGMP-independent manner.

• 대한약리학회지
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• 제32권2호
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• pp.283-292
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• 1996

The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region. In this study, single neurons of the spinal dorsal horn were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by patch clamp technique. Transverse slice ($(300{\mu}m$) of lumbar spinal cords from young rats$(7{\sim}14\;days)$ were sequentially treated with two pretenses(pronase 0.75 mg/ml and thermolysin 0.75 mg/ml), then single neurons were mechanically dissociated. These neurons showed near-intact morphology such as multipolar, ellipsoidal and bipolar, and pyramidal cells and we recorded the typical whole cell currents of $K^+$, $Ca^{2+}$ and ligand-operated channels from these neurons. Glutamate $(30{\mu}M)$ and N-methyl-D-aspartate(NMDA, $30{\mu}M)$ induced inward currents of $117{\pm}12.4$ pA(n=5) and $49{\pm}6.9$ pA(n=3), respectively. Glycine $(1{\mu}M)$ potentiated glutamate-induced currents $4{\sim}5$ times and NMDA-induced currents $8{\sim}10$ times. In addition, glycine $(30{\mu}M)$ induced Inward current ($31{\pm}6.1$ nA, n=2), which was rapidly desensitized after the peak to a new steady-state level. However, the inward currents induced by ${\gamma}-amino$ butyric acid(GABA, $1{\mu}M$) decreased continuously after the peak($226{\pm}41.6$ pA, n=3) under the similar experimental condition. The ionic currents and pharmacological responses of isolated neurons in this work were similar to those observed in vivo or in vitro spinal cord slice, indicating that acutely isolated neurons could be effectively used for further pharmacological studies.

• 생물정신의학
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• 제23권2호
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• pp.48-56
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• 2016

Alzheimer's disease (AD) is a neurodegenerative disorder in which neuronal loss causes cognitive decline and other neuropsychiatric problems. It can be diagnosed based on history, examination, and appropriate objective assessments, using standard criteria such as the Diagnostic and Statistical Manual of Mental Disorders or the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA). Brain imaging and biomarkers are making progress in the differential diagnoses among the different disorders. The cholinesterase inhibitors, donepezil, rivastigmine and galantamine and N-methyl-D-aspartate receptors antagonist memantine are approved by the US Food and Drug Administration for AD. Recently some acetylcholinesterase inhibitors gained approval for the treatment of severe AD and became available in a higher dose formulation or a patch formulation. Optimal care in AD is multifactorial and it should include early diagnosis and multidisciplinary care with pharmacological and nonpharmacological interventions including exercise interventions, cognitive interventions and maintenance of social networks.

• The Korean Journal of Physiology and Pharmacology
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• 제1권2호
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• pp.117-125
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• 1997

The N-methyl-D-aspartate (NMDA) receptor-mediated glutamatergic neurotransmission is involved in synaptic plasticity, developmental processes, learning and memory and many neuropathological disorders including age-related diseases. In the present study, regulation of the NMDA receptor properties by various ligands was investigated using $[^3H]MK-801$ binding studies in the synaptic membranes of young and aged rat forebrains. The binding in the presence of glutamate and glycine increased dramatically with growth between 1 and 6 weeks old, and thereafter declined gradually with aging. Glutamate, glycine or spermine respectively increased the binding with growth. Glutamate maintained the binding during aging, while glycine or spermine significantly decreased the binding in the aged brain. The maximum stimulation by glycine varied depending on the ages of brains. Greater sensitivity to glycine was observed at 1 week and 3 months and the sensitivity was significantly reduced in the aged brain. In contrast, spermine showed similar stimulation patterns in young and aged rats. These results indicated that the functional properties of the NMDA receptor-ion channel complex in young and aged rat forebrains are differentially regulated by agonists, and the reduction of the receptor function with normal aging may be, in some degree, due to the reduction of the receptor sensitivity to glycine.