• The Korean Journal of Pharmacology
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• v.32 no.1
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• pp.1-11
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• 1996

Since it has been reported that the depolarization-induced norepinephrine (NE) release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the NE release has not been clearly elucidated yet. Therefore, it was attempted to clarify the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of NE release in this study. Slices from rat hippocampus were equilibrated with $^3H-norepinephrine$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $Vcm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $1{\sim}30{\mu}M$, decreased the NE release in a dose-dependent manner, without affecting the basal rate of release. The adenosine effects were significantly inhibited by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, $2{\mu}M$), a selective $A_1-receptor$ antagonist. The responses to N-ethylmaleimide (NEM, 10 & $30{\mu}M$), a SH-alkylating agent of G-protein, were characterized by increments of the evoked NE-release and the basal release, and the adenosine effects were completely abolished by NEM pretreatment. $4{\beta}-Phorbol$ 12,13-dibutyrate (PDB, $1{\mu}M$), a specific protein kinase C (PKC) activator, increased the evoked NE release, whereas polymyxin B sulfate (PMB,0.1 mg), a PKC inhibitor, decreased the release, and the adenosine effects were inhibited by these agents. Nifedipine $(1{\mu}M)$, a $Ca^{2+}-channel$ blocker of dihydropyridine analogue, did not affect the adenosine effect. Tetraethylammonium (TEA, 3 mM) increased the evoked NE release, and inhibited the adenosine effects, but glibenclamide, a ATP dependent $K^+-channel$ blocker, did not. Finally, 8-bromo cyclic AMP (100 & $300{\mu}M$), a membrane-permeable analogue of cAMP, did not alter the NE release, but adenosine effects were inhibited by pretreatment with 8br-cAMP. These results suggest that the decrement of the evoked NE-release by $A_1-adenosine$ receptor is mediated by the C-protein, which is coupled to protein kinase C, adenylate cyclase system and TEA sensitive $K^+-channel$, and that nifedipine-sensitive $Ca^{2+}-channel$ and glibenclamide-sensitive $K^+-channel$ are not involved in this process.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Journal of Soil and Groundwater Environment
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• v.13 no.1
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• pp.1-12
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• 2008

The purpose of this study is to characterize the hydrogeochemical characteristics of hot spring waters and to interpret the source of noble gases and the geochemical environment of the hot spring waters distributed along the eastern area of the Korean peninsula. For this purpose, We carried out the chemical, stable isotopic and noble gas isotopic analyses for eleven hot spring water and fourteen hot spring gas samples collected from six hot spring sites. The hot spring waters except the Osaek hot spring water show the pH range of 7.0 to 9.1. However, the Osaek $CO_2$-rich hot spring water shows a weak acid of pH 5.7. The temperature of hot spring waters in the study area ranges from $25.7^{\circ}C$ to $68.3^{\circ}C$. Electrical conductivity of hot spring waters varies widely from 202 to $7,130{\mu}S/cm$. High electrical conductivity (av., $3,890{\mu}S/sm$) by high Na and Cl contents of the Haeundae and the Dongrae hot spring waters indicates that the hot spring waters were mixed with seawater in the subsurface thermal system. The type of hot springs in the viewpoint of dissolved components can be grouped into three types: (1) alkaline Na-$HCO_3$ type including sulfur gas of the Osaek, Baekam, Dukgu and Chuksan hot springs, and (2) saline Na-Cl type of the Haeundae and Dongrae hot springs, and (3) weak acid $CO_2$-rich Na-$HCO_3$ type of Osaek hot spring. Tritium ratios of the Haeundae and the Dongrae hot springs indicate different residence time in their aquifers of older water of $0.0{\sim}0.3$ TU and younger water of $5.9{\sim}8.8$ TU. The ${\delta}^{18}O$ and ${\delta}D$ values of hot spring waters indicate that they originate from the meteoric water, and that the values also reflect a latitude effect according to their locations. $^3He/^4He$ ratios of the hot spring waters except Osaek $CO_2$-rich hot spring water range from $0.1{\times}10^{-6}$ to $1.1{\times}10^{-6}$ which are plotted above the mixing line between air and crustal components. It means that the He gas in hot spring waters was originated mainly from atmosphere and crust sources, and partly from mantle sources. The Osaek $CO_2$-rich hot spring water shows $3.3{\times}10^{-6}$ in $^3He/^4He$ ratio that is 2.4 times higher than those of atmosphere. It provides clearly a helium source from the deep mantle. $^{40}Ar/^{36}Ar$ ratios of hot spring water are in the range of an atmosphere source.