• Title/Summary/Keyword: dopamine

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Dopamine determination using a biosensor based on multiwall carbon nanotubes paste and burley tobacco-peroxidase (담배 잎-peroxidase와 다중벽 탄소 나노튜브를 이용한 dopamine의 정량)

  • Kwon, Hyoshik;Jeon, Byong-Suk;Pak, Yongnam
    • Analytical Science and Technology
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    • v.28 no.2
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    • pp.98-105
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    • 2015
  • The development of an enzymatic biosensor for dopamine determination based on multiwall carbon nanotubes (MWCNTs) and peroxidase obtained from the crude extract of burley tobacco (Nicotiana tabacum L.) was proposed. Peroxidase catalyzes the oxidation of dopamine to dopamine quinone. The influence on the response of analytical parameters of biosensors such as enzyme concentration, dopamine concentration, pH, and phosphate buffer solution concentration were investigated. The analytical parameters obtained, including sensitivity, linearity, and stability, were investigated. The proposed method for dopamine determination presented good selectivity even in the presence of uric acid and ascorbic acid. The sensor presented a higher response for dopamine in 0.010 M phosphate buffer at pH 6.50, with an applied potential of -0.15 V. The detection limit of the electrode was 2.7×10−6 M (S/N = 3) and the relative standard deviation of the measurements, which were repeated 10 times using 5.0×10−2 M dopamine, was 1.3%.

Application of HPLC with Electrochemical Detection to Assaying Tyrosine Hydroxylase Activity and Dopamine Content in Dissociated Cultures of Fetal Rat Brainstem (흰쥐 태 뇌간의 세포배양에서 HPLC-전기화학검출을 이용한 Tyrosine Hydroxylase 활성 및 Dopamine의 정량)

  • Song, Dong-Keun;Wie, Myung-Bok;Park, Chan-Woong;Kim, Yung-Hi
    • The Korean Journal of Pharmacology
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    • v.27 no.1
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    • pp.7-12
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    • 1991
  • We measured the developmental increase of tyrosine hydroxylase(TH) activity and dopamine content with high performance liquid chromatography with electrochemical detection(HPLC-EC) in dissociated cultures of fetal rat brainstem(E14). TH activity and dopamine content increased progressively upto 7 days in vitro, when the effects of various drugs on the dopamine contents were studied. ${\alpha}-Methyl-p-tyrosine$, a TH inhibitor and NSD-1015, an inhibitor of aromatic amiono acid decarboxylase effectively depleted dopamine contents. Dopamine contents were depleted by reserpine and increased by pargyline. When cultures grown for 1 week in control medium were then exposed to tetrodotoxin$(0.1\;{\mu}M$) for 7 days, exposure to tetrodotoxin markedly decreased TH activity. All the above results indicate that dopamine metabolism in the cultered cells reflect reliably the property of brain dopamine metabolism. We suggest that measuring TH activity and dopamine content in brainstem culture with HPLC-EC can be useful tool in the study of pharmacology as well as toxicology of the central dopaminergic system.

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Effects of Harmaline and Harmalol on Dopamine Quinone-induced Brain Mitochondrial Dysfunction

  • Han, Eun-Sook;Lee, Chung-Soo
    • Biomolecules & Therapeutics
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    • v.10 no.3
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    • pp.152-158
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    • 2002
  • The present study elucidated the effect of $\beta$-carbolines (harmaline and harmalol) on brain mitochondlial dysfunction caused by the tyrosinase-induced oxidation of dopamine. Harmaline, harmalol and antioxidant enzymes (SOD and catalase) attenuated the dopamine-induced alteration of membrane potential, cytochrome c release and thiol oxidation in mitochondria. In contrast, antioxidant enzymes failed to reverse mitochondrial dysfunction induced by dopmnine plus tyrosinase. $\beta$-Carbolines decreased the damaging effect of dopamine plus tyrosinase against mitochondria, except no effect of harmalol on thiol oxidation. Antioxidant enzymes decreased the melanin formation from dopamine in the reaction mixture containing mitochondria but did not reduce the formation of dopamine quinone caused by tyrosinase. Both harmalol and harmaline inhibited the formation of reactive quinone and melanin. Harmalol being more effective for quinone formation and vise versa. The results indicate that compared to MAO-induced dopamine oxidation, the toxic effect of dopamine in the presence of tyrosinase against mitochondria may be accomplished by the dopamine quinone and toxic substances other than reactive oxygen species. $\beta$-Carbolines may decrease the dopamine plus tyrosinase-induced brain mitochondrial dysfunction by inhibition of the formation of reactive quinone and the change in membrane permeability.

Functional Characterization and Regional Expression of Dopamine Transporter (도파민 수송체의 기능적 특성 및 발현에 관한 연구)

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.161-168
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    • 1995
  • Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

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Effect of Dopamine on Propagation and Frequency of Slow Wave in Cat Isolated Stomach Muscle (적출한 고양이 위(胃) 근절편에서 서파의 전파 및 발생빈도에 미치는 Dopamine의 영향)

  • Jo, Yang-Hyeok;Sim, Sang-Soo;Kim, Myung-Suk;Kim, Chung-Chin;Choi, Hyun
    • The Korean Journal of Physiology
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    • v.20 no.1
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    • pp.9-15
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    • 1986
  • The effect of dopamine on the propagation and the frequency electrical activities (slow wave) of the stomach was studied in isolated stomach muscle strips of 145 cats, The gastric slow wave monopolarly recorded by four capillary electrodes (Ag-AgCl) in Krebs-Ringer solution $(ph\;7.4,\;temperature\;36{\pm}0.5^{\circ}C)$ bubbled with 5% $CO_2$ in $O_2$. Dopamine caused concentration-dependent changes of direction of slow wave propagation with decline in development of irregular propagation by domperidone pretreatment. Dopamine also increased the variation of slow wave frequency concentration-dependently. The variation of slow wave frequency induced by dopamine was significantly inhibited by pretreatment with domperidone and phentolamine but not with propranolol, hexamethonium and tetrodotoxin. It is therefore suggested that dopamine plays a role in the genesis of gastric electrical abnormality acting on dopamine receptors and partly on ${\alpha}-adrenergic$ receptors in cats.

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EFFECTS OF ACUTE AND SUBACUTE ADMINISTRATION OF COCAINE ON DOPAMINERGIC SYSTEMS IN THE RAT STRIATUM

  • Lim, D.K.;Ho, I.K.
    • Toxicological Research
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    • v.6 no.1
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    • pp.75-88
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    • 1990
  • The characteristics of dopamine uptake, D-1 and D-2 receptors after acute and subacute cocaine administration were determind in striatum from WKY and SHR. Cocaine was administered either acutely (40 mg/kg, s.c.) or twice daily (20 mg/kg, s.c.) for 3 and 7 days in 9-wk old WKY and SHR. Rats were sacrificed 30 min, 2 or 24 h after the single injection and 18 h after the last administration to the subacutely treated group. The changes in dopamine uptake, dopamine uptake sites, D-1 and D-2 receptors were determined using $(^3H)$dopamine, $(^3H)$-GBR-12935, $(^3H)$SCH-23390 and $(^3H)$sulpiride, respectively. In acutely treated rats, significant increases in $V_{max}$of dopamine uptake were observed 30 min after the cocanine injection in both strains without changes in $K_m$ values. The in vitro $IC_{50}$for cocaine was significantly decreased 30 min in WKY and 2 h in SHR. However, that for in vitro GBR-12909 was significantly increased 30 min and 2 h in both strains. Also densities of $(^3H)$-GBR-12935 binding sites were significantly increased 30 min and 2 h without changes in their $K_d$. Significant increases in D-2 receptor density were observed 30 min, 2 or 24 h after acute injection in both strains without changes in their affinities. The density of D-1 receptor was significantly decreased 30 min after the injection in WKY, but not in SHR. In subacutely treated rats, a significant increase in $K_m$ of dopamine uptake was observed in 7-day treated SHR. The in vitro $IC_{50}$fot GBR-12909 was significantly increased in 3-day treated WKY. The density of D-1 receptors was significantly increased in 3- and 7-day treated WKY, but not in SHR. The affinity of both binding sites remained unchanged. The results suggest that cocanine administration alters dopamine uptake, characteristics of dopamine uptake sites and dopamine receptor binding characteristics in rat brain. Furthermore, D-1 and D-2 dopamine receptors appear to be differently regulated.

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Role of Dopamine upon Spontaneous Contraction of Isolated Rat Uterus in Diestrus State (Dopamine이 흰쥐 자궁의 자발적 수축에 미치는 영향)

  • Park, Hyoung-Jin;Sim, Yeo-Rim;Jo, Yang-Hyeok
    • The Korean Journal of Physiology
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    • v.17 no.2
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    • pp.103-107
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    • 1983
  • This experiment was undertaken to see whether dopamine has any effect on a uterine function and whether the uterus has a dopamine receptor. We used 14 female rats in the diestrus state which was identified by a vaginal smear. Under ether anesthesia, 3 pieces(1 cm length) from each side of the uterus were dissected out and mounted in 3 tissue chambers (4 cm diameter, 10 cm height) that contained Krebs-Ringer solution. The solution was continuously aerated with 95% $O_2$ containing 5% $CO_2$ and kept $37^{\circ}C$ consistantly during the whole experimental period. The spontaneous contractile activity of the isolated uterus was recorded using a force transducer. After a recovery period of 15 min in the chamber, the following experiments were carried out. In 7 rats, each piece of the uterus was received dopamine at concentrations of $10^{-4}$, $10^{-5}$ or $10^{-6}\;M$ for 10 min and then followed by domperidone at a concentration of $10^{-5}\;M$. In another 7 rats, each piece was received domperidone, a specific peripheral dopamine receptor antagonist, was administered at a concentration of $10^{-5}\;M$ for 5 min prior to dopamine at concentrations of $10^{-4}$, $10^{-5}$, or $10^{-6}\;M$. Dopamine inhibited the spontaneous uterine contraction dose-dependently (r=0.99, p<.01). The inhibited contractility by dopomine was significantly (P<.05) resumed by post-treatment of domperidone. Pre-treatment of domperidone also blocked significantly(p<.05) the inhibitory effect of dopamine. It is concluded from these results that dopamine has inhibitory role upon the spontaneous uterine contraction of the rat in the diestrus state and domperidone antagonized the inhibitory effect of dopamine. These results suggest strongly that dopamine may exert the inhibitory effect via the dopamine receptor in the rat uterus.

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Effect of Dopamine, SKF 81297, a Dopamine D$_1$-Receptor Agonist and TNPA, a Dopamine D$_2$-Receptor Agoinst on the Blood Pressure in Rats (Dopamine, Dopamine D$_1$-Receptor 효능제인 SKF 81297 및 Dopamine, D$_2$-Receptor 효능제인 TNPA의 흰쥐 혈압에 대한 영향)

  • Ko, Suk-Tai;Lim, Dong-Yoon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.97-97
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    • 2001
  • This Study was attempted to investigate tile effect of dopamine, SKF 81297, a dopamine D$_1$-receptor agonist, and TNPA, a dopamine D$_2$-receptor agonist, on the blood pressure in rat. Dopamine exhibited the hypertensive action in proportion to the doses of 1.0, 3.0 arid 10.0 $\mu\textrm{g}$/kg i.v., these hypertensive action of dopamine was blocked significantly by SCH 23390, a dopamine D$_1$-receptor antagonist, on the other hand, more potentiated by raclopride, a dopamine D$_1$-receptor antagonist. SKF 81297 produced hypertensive action in a dose of 1.0 $\mu\textrm{g}$/kg i.v., wherease hypotensive action in proportion to administered doses 3.0 and 10.0 $\mu\textrm{g}$/kg i.v., these hypertensive action of SKF 81297 in a dose of 1.0 $\mu\textrm{g}$/kg i.v. was not influenced by SCH 23390 or raclopride, but hypotensive action of SKF 81297 in tile doses of 3.0 and 10.0 $\mu\textrm{g}$/kg i.v. was weakened significantly by SCH 23390, but more strenthened by raclopride. TNPA showed the hypotensive action in inverse proportion to administered doses of 1.0, 3.0 and 10.0 $\mu\textrm{g}$/kg i.v., these hypotensive action was reversed to hypertensive action in inverse proportion to the administered doses of TNPA by SCH 23390 and raclopride.

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The Relationship between Taq I A Dopamine $D_2$ Receptor Polymorphism and Therapeutic Response to Antipsychotics in Schizophrenic Patients (정신분열병환자에서 Taq I A 도파민 $D_2$ 수용체 다형성과 항정신병약물의 치료반응과의 연관)

  • Kang, Cheol Joong
    • Korean Journal of Biological Psychiatry
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    • v.12 no.2
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    • pp.107-113
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    • 2005
  • Purpose:In an attempt to predict the interpersonal differences of therapeutic response to antipsychotic drugs on pharmaco-genetic bases, this study was designed to investigate the relationship between the therapeutic response to antipsychotic drugs and Taq I A dopamine $D_2$ receptor polymorphism in schizophrenic patients. Methods:The subjects were 158 patients diagnosed with schizophrenia(DSM-IV). The therapeutic response to antipsychotic drugs was evaluated using the Treatment Response Scale(TRS) retrospectively. Patients were divided into two groups, dopamine receptor antagonist responders, and serotonin-dopamine antagonist responders. The patients' Taq I A dopamine $D_2$ receptor polymorphism was determined by polymerase chain reaction(PCR) and restriction fragment length polymorphism(RFLP). Results:The dopamine receptor antagonist responders had the A1 allele in significantly higher incidences (${\chi}^2$(1)=4.875, p=0.027, two-tailed). No significant difference was found among the serotonin-dopamine antagonist responders between those with or without the A1 allele. Conclusions:The patients with the A1 allele responded better to dopamine receptor antagonists than those with no A1 allele. Based on these results, it is suggested that the pharmacological effect of dopamine receptor antagonists can be predicted depending on the presence of the A1 allele in schizophrenic patients.

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Differential Effect of Harmalol and Deprenyl on Dopamine-Induced Mitochondrial Membrane Permeability Change in PC12 Cells

  • Lee, Chung-Soo
    • Biomolecules & Therapeutics
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    • v.12 no.1
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    • pp.9-18
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    • 2004
  • Opening of the mitochondrial permeability transition pore has been recognized to be involved in cell death. The present study investigated the effect of ${\beta}$-carbolines (harmaline and harmalol) and deprenyl on the dopamine-induced change in the mitochondrial membrane permeability and cell death in differentiated PC12 cells. Cell death due to 250 4{\mu}$M dopamine was inhibited by caspase inhibitors (z-IETD.fmk, z-LEHD.fmk and z-DQMD.fmk) and antioxidants (N-acetylcysteine, ascorbate, superoxide dismutase, catalase and carboxy-PTIO). ${\beta}$-Carbolines prevented the dopamine-induced cell death in PCl2 cells, while deprenyl did not inhibit cell death. ${\beta}$-Carbolines decreased the condensation and fragmentation of nuclei caused by dopamine in PC12 cells. ${\beta}$-Carbolines inhibited the decrease in mitochondrial transmembrane potential, cytochrome c release, formation of reactive oxygen species and depletion of GSH caused by dopamine in PC12 cells, whereas deprenyl did not decrease dopamine-induced mitochondrial damage. ${\beta}$-Carbolines, deprenyl and antioxidants depressed the formation of nitric oxide and melanin in dopamine-treated PC12 cells. The results suggest that cell death due to dopamine PC12 cells is mediated by caspase-8, -9 and -3. Unlike deprenyl, ${\beta}$-carbolines may attenuate the dopamineinduced cell death in PC12 cells by suppressing change in the mitochondrial membrane permeability through inhibition of the toxic action of reactive oxygen and nitrogen species.