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김용환,차인호,조현호,김상현,진형근,김영일,김도 한국수의공중보건학회 1996 예방수의학회지 Vol.20 No.1
Identification of ATP-sensitive $K^+$ Conductances in Male Rat Major Pelvic Ganglion Neurons
Park, Kyu-Sang,Cha, Seung-Kyu,Lee, Keon-Il,Jun, Jae-Yeoul,Jeong, Seong-Woo,Kong, In-Deok,Lee, Joong-Woo The Korean Society of Pharmacology 2002 The Korean Journal of Physiology & Pharmacology Vol.6 No.5
Major pelvic ganglia (MPG) neurons are classified into sympathetic and parasympathetic neurons according to the electrophysiological properties; membrane capacitance (Cm), expression of T-type $Ca^{2+}$ channels, and the firing patterns during depolarization. In the present study, function and molecular expression of ATP-sensitive $K^+\;(K_{ATP})$ channels was investigated in MPG neurons of male rats. Only in parasympathetic MPG neurons showing phasic firing patterns, hyperpolarizing changes were elicited by the application of diazoxide, an activator of $K_{ATP}$ channels. Glibenclamide $(10{\mu}M),$ a $K_{ATP}$ channel blocker, completely abolished the diazoxide-induced hyperpolarization. Diazoxide increased inward currents at high $K^+$ (90 mM) external solution, which was also blocked by glibenclamide. The metabolic inhibition by the treatment with mitochondrial respiratory chain inhibitors (rotenone and antimycin) hyperpolarized the resting membrane potential of parasympathetic neurons, which was not observed in sympathetic neurons. The hyperpolarizing response to metabolic inhibition was partially blocked by glibenclamide. RT-PCR analysis revealed that MPG neurons mainly expressed the $K_{ATP}$ channel subunits of Kir6.2 and SUR1. Our results suggest that MPG neurons have $K_{ATP}$ channels, mainly formed by Kir6.2 and SUR1, with phenotype-specificity, and that the conductance through this channel in parasympathetic neurons may contribute to the changes in excitability during hypoxia and/or metabolic inhibition.
Verocytotoxin 생성 대장균의 분포 및 Receptor를 이용한 RELISA 법의 개발 I. 국내 가축에 대한 생성 대장균의 분포조사 및 Hybrid Toxin의 조합
김용환(Yong Hwan Kim),차인호(In Ho Cha),조현호(Hyun Ho Cho),김상현(Sang Hyun Kim),진형근(Hyeung Keon Jin),김영일(Young Il Kim),김도(Do Kim) 한국예방수의학회 1996 예방수의학회지 Vol.20 No.1
Verocytotoxin-producing Escherichia coli(VTEC) as etiological agents of diarrhea represents one of the most recent observations in the field of enteric infections. In epidemiological survey related to VTEC infection of cattle and pigs in Korea, a total of 675 stools of cattle and 784 stools of pigs collected at slaughterhouse were examined for the presence of VTEC. The isolation rates of VTEC from cattle and pigs were 14.4% and 11.4%, respectively. Particularly, 2 strains of E. coli O157:H7 were isolated from cattle. Cytotoxicity for culture supernatants of isolates was assayed to Vero and HeLa cell monolayers. The level of cytotoxin produced by isolated strains were 102~107 cytotoxic dose (CD50)/ml of incubated supernatants.
Characterization of Acetylcholine-induced Currents in Male Rat Pelvic Ganglion Neurons
Park, Joong-Hyun,Park, Kyu-Sang,Cha, Seung-Kyu,Lee, Keon-Il,Kim, Min-Jung,Park, Jong-Yeon,Kong, In-Deok,Lee, Joong-Woo The Korean Society of Pharmacology 2004 The Korean Journal of Physiology & Pharmacology Vol.8 No.4
The pelvic ganglia provide autonomic innervations to the various urogenital organs, such as the urinary bladder, prostate, and penis. It is well established that both sympathetic and parasympathetic synaptic transmissions in autonomic ganglia are mediated mainly by acetylcholine (ACh). Until now, however, the properties of ACh-induced currents and its receptors in pelvic ganglia have not clearly been elucidated. In the present study, biophysical characteristics and molecular nature of nicotinic acetylcholine receptors (nAChRs) were studied in sympathetic and parasympathetic major pelvic ganglion (MPG) neurons. MPG neurons isolated from male rat were enzymatically dissociated, and ionic currents were recorded by using the whole cell variant patch clamp technique. Total RNA from MPG neuron was prepared, and RT-PCR analysis was performed with specific primers for subunits of nAChRs. ACh dose-dependently elicited fast inward currents in both sympathetic and parasympathetic MPG neurons $(EC_{50};\;41.4\;{\mu}M\;and\;64.0\;{\mu}M,\;respectively)$. ACh-induced currents showed a strong inward rectification with a reversal potential near 0 mV in current-voltage relationship. Pharmacologically, mecamylamine as a selective antagonist for ${\alpha}3{\beta}4$ nAChR potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons $(IC_{50};\;0.53\;{\mu}M\;and\;0.22\;{\mu}M,\;respectively)$. Conversely, ${\alpha}-bungarotoxin$, ${\alpha}-methyllycaconitine$, and $dihydro-{\beta}-erythroidine$, which are known as potent and sensitive blockers for ${\alpha}7$ or ${\alpha}4{\beta}2$ nAChRs, below micromolar concentrations showed negligible effect. RT-PCR analysis revealed that ${\alpha}3$ and ${\beta}4$ subunits were predominantly expressed in MPG neurons. We suggest that MPG neurons have nAChRs containing ${\alpha}3$ and ${\beta}4$ subunits, and that their activation induces fast inward currents, possibly mediating the excitatory synaptic transmission in pelvic autonomic ganglia.
Identification of ATP-sensitive K<SUP></SUP> Conductances in Male Rat Major Pelvic Ganglion Neurons
Kyu-Sang Park,Seung-Kyu Cha,Keon-Il Lee,Jae Yeoul Jun,Seong-Woo Jeong,In Deok Kong,Joong Woo Lee 대한생리학회-대한약리학회 2002 The Korean Journal of Physiology & Pharmacology Vol.6 No.5
<P> Major pelvic ganglia (MPG) neurons are classified into sympathetic and parasympathetic neurons according to the electrophysiological properties; membrane capacitance (Cm), expression of T-type Ca<SUP>2</SUP> channels, and the firing patterns during depolarization. In the present study, function and molecular expression of ATP-sensitive K<SUP></SUP> (K<SUB>ATP</SUB>) channels was investigated in MPG neurons of male rats. Only in parasympathetic MPG neurons showing phasic firing patterns, hyperpolarizing changes were elicited by the application of diazoxide, an activator of K<SUB>ATP</SUB> channels. Glibenclamide (10μM), a K<SUB>ATP</SUB> channel blocker, completely abolished the diazoxide-induced hyperpolarization. Diazoxide increased inward currents at high K<SUP></SUP> (90 mM) external solution, which was also blocked by glibenclamide. The metabolic inhibition by the treatment with mitochondrial respiratory chain inhibitors (rotenone and antimycin) hyperpolarized the resting membrane potential of parasympathetic neurons, which was not observed in sympathetic neurons. The hyperpolarizing response to metabolic inhibition was partially blocked by glibenclamide. RT-PCR analysis revealed that MPG neurons mainly expressed the K<SUB>ATP</SUB> channel subunits of Kir6.2 and SUR1. Our results suggest that MPG neurons have K<SUB>ATP</SUB> channels, mainly formed by Kir6.2 and SUR1, with phenotype-specificity, and that the conductance through this channel in parasympathetic neurons may contribute to the changes in excitability during hypoxia and/or metabolic inhibition.
김용환(Yong Hwan Kim),차인호(In Ho Cha),조현호(Hyun Ho Cho),김상현(Sang Hyun Kim),진형근(Hyeung Keon Jin),김영일(Young Il Kim),김도(Do Kim) 한국예방수의학회 1996 예방수의학회지 Vol.20 No.1
Escherichia coli-derived verotoxins have been implicated in the etiology of the hemolytic uremic syndrome and hemorrhagic colitis in human, and edema disease in swine. Currently available toxin assays, however, are not suitable for most clinical or public health laboratories. This study was carried out to develop more sensitive and practical method for detection of low level verotoxins.
Characterization of Acetylcholine-induced Currents in Male Rat Pelvic Ganglion Neurons
Joong-Hyun Park,Kyu-Sang Park,Seung-Kyu Cha,Keon-Il Lee,Min-Jung Kim,Jong-Yeon Park,In Deok Kong,Joong-Woo Lee 대한생리학회-대한약리학회 2004 The Korean Journal of Physiology & Pharmacology Vol.8 No.4
The pelvic ganglia provide autonomic innervations to the various urogenital organs, such as the urinary bladder, prostate, and penis. It is well established that both sympathetic and parasympathetic synaptic transmissions in autonomic ganglia are mediated mainly by acetylcholine (ACh). Until now, however, the properties of ACh-induced currents and its receptors in pelvic ganglia have not clearly been elucidated. In the present study, biophysical characteristics and molecular nature of nicotinic acetylcholine receptors (nAChRs) were studied in sympathetic and parasympathetic major pelvic ganglion (MPG) neurons. MPG neurons isolated from male rat were enzymatically dissociated, and ionic currents were recorded by using the whole cell variant patch clamp technique. Total RNA from MPG neuron was prepared, and RT-PCR analysis was performed with specific primers for subunits of nAChRs. ACh dose-dependently elicited fast inward currents in both sympathetic and parasympathetic MPG neurons (EC<SUB>50</SUB>; 41.4μM and 64.0μM, respectively). ACh-induced currents showed a strong inward rectification with a reversal potential near 0 mV in current-voltage relationship. Pharmacologically, mecamylamine as a selective antagonist for α3β4 nAChR potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons (IC<SUB>50</SUB>; 0.53μM and 0.22μM, respectively). Conversely, α- bungarotoxin, α-methyllycaconitine, and dihydro-β-erythroidine, which are known as potent and sensitive blockers for α7 or α4β2 nAChRs, below micromolar concentrations showed negligible effect. RT-PCR analysis revealed that α3 and β4 subunits were predominantly expressed in MPG neurons. We suggest that MPG neurons have nAChRs containing α3 and β4 subunits, and that their activation induces fast inward currents, possibly mediating the excitatory synaptic transmission in pelvic autonomic ganglia.