5-HT 흡수억제성 항우울제들이 가토혈소판의 [$^3H$]Imipramine과 [$^3H$]Paroxetine Binding, [$^3H$]5-HT 흡수, 및 5-HT함량에 미치는 영향

원경식,이민수,신경호,전보권,곽동일,Won, Kyong-Sik,Lee, Min-Soo,Shin, Kyung-Ho,Chun, Boe-Gwun,Kwak, Dong-Il 대한생물정신의학회 1994 생물정신의학 Vol.1 No.1

Many evidences are compatible with the correlation between the inhibition of [$^3H$] imipramine([$^3H$]IMI) and [$^3H$]paroxetine([$^3H$]PAT) binding to the 5-hydroxytryptamine(5-HT) transporter complex and the 5-HT uptake of 5-HT neurons and platelets, and most antidepressants have been shown to inhibit the [$^3H$]IMI and [$^3H$]PAT binding and the neuronal 5-HT uptake. However, several paradoxical research findings led to doubt about the pharmacological significance of the [$^3H$]IMI and [$^3H$]PAT binding sites. This study was carried to clarify the correlation between the [$^3H$]IMI and [$^3H$]PAT binding parameters and the tissue 5-HT content or/and [$^3H$]5-HT uptake in the rabbit platelet, which contains 40 times ad much 5-HT as that of human platelet and shows the 10 fold higher $B_{max}$ of the 5-HT transporter binding to a 5-HT uptake inhibitor. The rabbits were treated for 28 days with amitriptyline(4mg/kg/day : AP), fluoxetine(0.5mg/kg/day : FO), and sertraline(0.5mg/kg/day : SA) via an Alzet osmotic pump implanted for constant infusion. The [$^3H$]IMI binding $B_{max}$ and $K_d$ of the rabbit platelets were $6.4{\pm}1.2$pmol/mg protein and $10.9{\pm}2.1$nM and those in the [$^3H$]PAT binding were $8.6{\pm}1.1$pmol/mg protein and $1.6{\pm}0.3$nM, respectively. AP slightly increased $B_{max}$ of [$^3H$]IMI binding and both [$^3H$]IMI binding and [$^3H$]PAT binding $K_d$, and i contrast, it slightly decreased $B_{max}$ of [$^3H$]PAT binding. FO Slightly increased $K_d$ of both and [$^3H$]IMI and [$^3H$]PAT binding and slightly decreased $B_{max}$ of [$^3H$]IMI and [$^3H$]PAT binding. SA produced the significant increase of [$^3H$]PAT binding $B_{max}$ and the slight increase of both [$^3H$]IMI and [$^3H$]PAT binding $K_d$ and in contrast, it slightly decreased $B_{max}$ and of [$^3H$]IMI binding. And, the $V_{max}$ and $K_m$ of platelet [$^3H$]5-HT uptake were $24.2{\pm}2.4$pmol/$10^8$ platelets/min and $3.3{\pm}0.3$nM, respectively. The $V_{max}$ was little affected by AP, FO, or SA, but the [$^3H$]5-HT uptake $K_m$ value was moderately increased by FO. However, the platelet 5-HT content was moderately decreased by all of the 5-HT uptake inhibitors used in this study. These results seem to be consistent with the allosterical and competitive interaction of 5-HT uptake inhibiting antidepressants with each other as well as 5-HT in the 5-HT transporter binding, and provide no support for the view that the potencies of 5-HT uptake inhibitors to inhibit the [$^3H$]IMI or [$^3H$]PAT binding with 5-HT transporter complex correlate with their antidepressant potencies.

Lamotrigine, an antiepileptic drug, inhibits 5-HT3 receptor currents in NCB-20 neuroblastoma cells

김기정,전승현,성기욱 대한약리학회 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.2

Lamotrigine is an antiepileptic drug widely used to treat epileptic seizures. Using whole-cell voltage clamp recordings in combination with a fast drug application approach, we investigated the effects of lamotrigine on 5-hydroxytryptamine (5-HT)3 receptors in NCB-20 neuroblastoma cells. Coapplication of lamotrigine (1~300 mM) resulted in a concentration-dependent reduction in peak amplitude of currents induced by 3 mM of 5-HT for an IC50 value of 28.2±3.6 mM with a Hill coefficient of 1.2±0.1. These peak amplitude decreases were accompanied by the rise slope reduction. In addition, 5-HT3-mediated currents evoked by 1 mM dopamine, a partial 5-HT3 receptor agonist, were inhibited by lamotrigine co-application. The EC50 of 5-HT for 5-HT3 receptor currents were shifted to the right by co-application of lamotrigine without a significant change of maximal effect. Currents activated by 5-HT and lamotrigine co-application in the presence of 1 min pretreatment of lamotrigine were similar to those activated by 5-HT and lamotrigine co-application alone. Moreover, subsequent application of lamotrigine in the presence of 5-HT and 5-hydroxyindole, known to attenuate 5-HT3 receptor desensitization, inhibited 5-HT3 receptor currents in a concentration-dependent manner. The deactivation of 5-HT3 receptor was delayed by washing with an external solution containing lamotrigine. Lamotrigine accelerated the desensitization process of 5-HT3 receptors. There was no voltage-dependency in the inhibitory effects of lamotrigine on the 5-HT3 receptor currents. These results indicate that lamotrigine inhibits 5-HT3-activated currents in a competitive manner by binding to the open state of the channels and blocking channel activation or accelerating receptor desensitization.

Lamotrigine, an antiepileptic drug, inhibits 5-HT₃ receptor currents in NCB-20 neuroblastoma cells

Kim, Ki Jung,Jeun, Seung Hyun,Sung, Ki-Wug The Korean Society of Pharmacology 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.2

Lamotrigine is an antiepileptic drug widely used to treat epileptic seizures. Using whole-cell voltage clamp recordings in combination with a fast drug application approach, we investigated the effects of lamotrigine on 5-hydroxytryptamine $(5-HT)_3$ receptors in NCB-20 neuroblastoma cells. Co-application of lamotrigine ($1{\sim}300{\mu}M$) resulted in a concentration-dependent reduction in peak amplitude of currents induced by $3{\mu}m$ of 5-HT for an $IC_{50}$ value of $28.2{\pm}3.6{\mu}M$ with a Hill coefficient of $1.2{\pm}0.1$. These peak amplitude decreases were accompanied by the rise slope reduction. In addition, $5-HT_3$-mediated currents evoked by 1 mM dopamine, a partial $5-HT_3$ receptor agonist, were inhibited by lamotrigine co-application. The $EC_{50}$ of 5-HT for $5-HT_3$ receptor currents were shifted to the right by co-application of lamotrigine without a significant change of maximal effect. Currents activated by 5-HT and lamotrigine co-application in the presence of 1 min pretreatment of lamotrigine were similar to those activated by 5-HT and lamotrigine co-application alone. Moreover, subsequent application of lamotrigine in the presence of 5-HT and 5-hydroxyindole, known to attenuate $5-HT_3$ receptor desensitization, inhibited $5-HT_3$ receptor currents in a concentration-dependent manner. The deactivation of $5-HT_3$ receptor was delayed by washing with an external solution containing lamotrigine. Lamotrigine accelerated the desensitization process of $5-HT_3$ receptors. There was no voltage-dependency in the inhibitory effects of lamotrigine on the $5-HT_3$ receptor currents. These results indicate that lamotrigine inhibits $5-HT_3$-activated currents in a competitive manner by binding to the open state of the channels and blocking channel activation or accelerating receptor desensitization.

Lamotrigine, an antiepileptic drug, inhibits 5-HT₃ receptor currents in NCB-20 neuroblastoma cells

Ki Jung Kim,Seung Hyun Jeun,Ki-Wug Sung 대한생리학회-대한약리학회 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.2

Lamotrigine is an antiepileptic drug widely used to treat epileptic seizures. Using whole-cell voltage clamp recordings in combination with a fast drug application approach, we investigated the effects of lamotrigine on 5-hydroxytryptamine (5-HT)₃ receptors in NCB-20 neuroblastoma cells. Coapplication of lamotrigine (1~300 μM) resulted in a concentration-dependent reduction in peak amplitude of currents induced by 3 μM of 5-HT for an IC₅₀ value of 28.2±3.6 μM with a Hill coefficient of 1.2±0.1. These peak amplitude decreases were accompanied by the rise slope reduction. In addition, 5-HT₃-mediated currents evoked by 1 mM dopamine, a partial 5-HT₃ receptor agonist, were inhibited by lamotrigine co-application. The EC₅₀ of 5-HT for 5-HT₃ receptor currents were shifted to the right by co-application of lamotrigine without a significant change of maximal effect. Currents activated by 5-HT and lamotrigine co-application in the presence of 1 min pretreatment of lamotrigine were similar to those activated by 5-HT and lamotrigine co-application alone. Moreover, subsequent application of lamotrigine in the presence of 5-HT and 5-hydroxyindole, known to attenuate 5-HT₃ receptor desensitization, inhibited 5-HT₃receptor currents in a concentration-dependent manner. The deactivation of 5-HT₃ receptor was delayed by washing with an external solution containing lamotrigine. Lamotrigine accelerated the desensitization process of 5-HT₃ receptors. There was no voltage-dependency in the inhibitory effects of lamotrigine on the 5-HT₃ receptor currents. These results indicate that lamotrigine inhibits 5-HT₃-activated currents in a competitive manner by binding to the open state of the channels and blocking channel activation or accelerating receptor desensitization.

백서 대뇌 피질 절편에서 허혈에 의한 [³H]Hydroxytryptamine의 유리 기전에 관한 연구

송대원,김영현,김기원 대한신경과학회 1997 대한신경과학회지 Vol.15 No.5

Experimental Research Article : Amino acid residues involved in agonist binding and its Linking to channel gating, proximal to transmembrane domain of 5-HT3A receptor for halothane modulation

( Mi Kyeong Kim ),( Kyeong Tae Min ),( Bon Nyeo Koo ) 대한마취과학회 2009 Korean Journal of Anesthesiology Vol.56 No.1

Background: The 5-hydroxytryptamine type 3 (5-HT3) receptor is a member of the Cys-loop superfamily of ligand-gated ion channels (LGICs) and modulated by pharmacologic relevant concentrations of volatile anesthetics or n-alcohols like most receptors of LGICs. The goal of this study was to reveal whether the site-directed single mutations of E-106, F-107 and R-222 in 5-HT3 receptor may affect the anesthetic modulation of halothane known as positive modulator. Methods: The wild-type and mutant receptors, E106D, F107Y, R222F, R222V, were expressed in Xenopus Laevis oocytes and receptor function was assessed using two electrode voltage clamp techniques. Results: E106D, F107Y, R222F, R222V mutant 5-HT3A receptors were functionally expressed. F107Y mutant 5-HT3A receptors displayed decreased sensitivity to 5-HT compared to the wild type 5-HT3A receptor (P<0.05). Halothane showed positive modulation in both wild and F107Y mutant 5-HT3A receptors but F107Y mutant 5-HT3 receptor showed greater enhancing modulation comparing to wild-type receptor. Meanwhile, R222F and R222V mutant 5-HT3 receptor lost positive modulation with 1 and 2 MAC of halothane. Most interestingly, positive modulation by halothane was converted into negative modulation in E106D mutant 5-HT3A receptor. Conclusions: The present study implicate the amino acid residues known for agonist binding and linking agonist binding to channel gating might also have important role for anesthetic modulation in 5-HT3A receptor. (Korean J Anesthesiol 2009;56:66~73)

Mutations of Arginine 222 in Pre-transmembrane Domain I of Mouse 5-HT_3A Receptor Abolish 20(<i>R</i>)- But Not 20(<i>S</i>)-Ginsenoside Rg₃ Inhibition of 5-HT-Mediated Ion Currents

Lee, Byung-Hwan,Lee, Jun-Ho,Yoon, In-Soo,Lee, Joon-Hee,Choi, Sun-Hye,Shin, Tae-Joon,Pyo, Mi Kyung,Choi, Woo-Sung,Lee, Sang-Mok,Lim, Yoongho,Rhim, Hyewhon,Nah, Seung-Yeol Pharmaceutical Society of Japan 2007 BIOLOGICAL & PHARMACEUTICAL BULLETIN Vol.30 No.9

<P>Ginsenosides, active ingredients of <I>Panax</I> ginseng, exist as stereoisomers depending on the position of the hydroxyl group on carbon-20; <I>i.e.</I> 20(<I>R</I>)-ginsenoside and 20(<I>S</I>)-ginsenoside are epimers. We previously investigated the structure–activity relationship of the ginsenoside Rg<SUB>3</SUB> stereoisomers, 20-<I>R</I>-protopanaxatriol-3-[<I>O</I>-β-<SMALL>D</SMALL>-glucopyranosyl (1→2)-β-glucopyranoside], (20(<I>R</I>)-Rg<SUB>3</SUB>) and 20-<I>S</I>-protopanaxatriol-3-[<I>O</I>-β-<SMALL>D</SMALL>-glucopyranosyl (1→2)-β-glucopyranoside], (20(<I>S</I>)-Rg<SUB>3</SUB>) in regulating 5-HT<SUB>3A</SUB> receptor-mediated ion currents (<I>I</I><SUB>5-HT</SUB>) expressed in <I>Xenopus</I> oocytes and found that 20(<I>S</I>)-Rg<SUB>3</SUB> rather than 20(<I>R</I>)-Rg<SUB>3</SUB> was more stronger inhibitor of <I>I</I><SUB>5-HT</SUB>. In the present study, we further investigated the effects of 20(<I>R</I>)-Rg<SUB>3</SUB> and 20(<I>S</I>)-Rg<SUB>3</SUB> on mouse 5-HT<SUB>3A</SUB> receptor channel activity after site-directed mutations of 5-HT<SUB>3A</SUB> receptor facilitation site, which is located at pre-transmembrane domain I (pre-TM1). 5-HT<SUB>3A</SUB> receptor was expressed in <I>Xenopus</I> oocytes, and <I>I</I><SUB>5-HT</SUB> was measured using two-electrode voltage clamp technique. In wild-type, both 20(<I>R</I>)-Rg<SUB>3</SUB> and 20(<I>S</I>)-Rg<SUB>3</SUB> inhibited <I>I</I><SUB>5-HT</SUB> with concentration-dependent and reversible manner. Induction of 5-HT<SUB>3A</SUB> receptor facilitation by point mutations of pre-TM1 amino acid residue R222 to R222A, R222D, R222E or R222T not only decreased EC<SUB>50</SUB> values for <I>I</I><SUB>5-HT</SUB> compared to wild-type but also abolished 20(<I>R</I>)-Rg<SUB>3</SUB>-induced inhibition of <I>I</I><SUB>5-HT</SUB>. Those mutations also shifted the IC<SUB>50</SUB> values by 20(<I>S</I>)-Rg<SUB>3</SUB> into right direction by 2- to 4-folds compared with wild-type. These results indicate that 5-HT<SUB>3A</SUB> receptor facilitation differentially affects 20(<I>R</I>)-Rg<SUB>3</SUB>- and 20(<I>S</I>)-Rg<SUB>3</SUB>-mediated 5-HT<SUB>3A</SUB> receptor channel regulation.</P>

Serotoninergic modulation of GABAergic synaptic transmission in developing rat CA3 pyramidal neurons

Choi, In-Sun,Cho, Jin-Hwa,Kim, Jung-Tak,Park, Eun-Joo,Lee, Maan-Gee,Shin, Hong-In,Choi, Byung-Ju,Jang, Il-Sung Raven Press [etc.] 2007 Journal of neurochemistry Vol.103 No.6

<P>Abstract</P><P>Serotoninergic modulation of GABAergic mIPSCs was investigated in immature (postnatal 12–16-days old) rat CA3 pyramidal neurons using a conventional whole-cell patch clamp technique. Serotonin or 5-hydroxytryptamine (5-HT) (10 &mgr;mol/L) transiently and explosively increased mIPSC frequency with a small increase in the current amplitude. However, 5-HT did not affect the GABA-induced postsynaptic currents, indicating that 5-HT acts presynaptically to facilitate the probability of spontaneous GABA release. The 5-HT action on GABAergic mIPSC frequency was completely blocked by 100 nmol/L MDL72222, a selective 5-HT<SUB>3</SUB> receptor antagonist, and mimicked by mCPBG, a selective 5-HT<SUB>3</SUB> receptor agonist. The 5-HT action on GABAergic mIPSC frequency was completely occluded either in the presence of 200 &mgr;mol/L Cd<SUP>2+</SUP> or in the Na<SUP>+</SUP>-free external solution, suggesting that the 5-HT<SUB>3</SUB> receptor-mediated facilitation of mIPSC frequency requires a Ca<SUP>2+</SUP>influx passing through voltage-dependent Ca<SUP>2+</SUP>channels from the extracellular space, and that presynaptic 5-HT<SUB>3</SUB> receptors are less permeable to Ca<SUP>2+</SUP>. The 5-HT action on mIPSC frequency in the absence or presence of extracellular Na<SUP>+</SUP> gradually increased with postnatal development. Such a developmental change in the 5-HT<SUB>3</SUB> receptor-mediated facilitation of GABAergic transmission would play important roles in the regulation of excitability as well as development in CA3 pyramidal neurons.</P>

백서 대뇌 피질 절편에서 허혈에 의한 [3H]Hydroxytryptamine의 유리 기전에 관한 연구

송대원,김영현,김기원 대한신경과학회 1997 대한신경과학회지 Vol.15 No.5

Synthesis and SAR of N-Chlorophenyl Substituted Piperrazinylethyl-aminomethylpyrazoles as 5-HT_3A Inhibitors

Lee, Byung-Hwan,Choi, In-Sung,Rhim, Hye-Whon,Choi, Kyung-Il,Nah, Seung-Yeol,Nam, Ghil-Soo Korean Chemical Society 2009 Bulletin of the Korean Chemical Society Vol.30 No.11

5-$HT_{3}$ receptor;5-$HT_{3A}$ receptor channel activity;Novel 5-$HT_{3}$ receptor channel current blockers;Chlorophenyl substituted piperazinylethylaminomethylpyrazoles; The 5-$HT_{3A}$ receptors are one of ligand-gated ion channels and are known to be involved in visceral pain, anxiety, or anticancer agent-induced nausea and vomiting. In present study, we designed novel skeletons based on the developed 5-$HT_{3}$ receptor antagonists and evaluated their effects on 5-$HT_{3A}$ receptor channel currents ($I_{5-HT}$) of a series of pyrazole derivatives having N-chlorophenylpiperazine functionality (6-9). We found that most of N-p-chlorophenyl substituted piperazinyl-pyrazole derivatives (7b, 7c, 7e and 7h) exhibited the high potency for the inhibition of $I_{5-HT}$, whereas the compound without chloride (6) or with m-chlorophenyl group (a serious of 8 and 9) showed the low potency. These result indicate that p-chlorophenyl group is might play an important role for increasing the inhibitory potency on $I_{5-HT}$.