1.Effects of Fetal Mesencephalic Cell Grafts on the Intrastriatal 6-hydroxydoapmine Lesioned Rats.
Wan Seok JOO ; Eunjoo NAM ; Heh In IM ; Jin Ah JUNG ; Eun Sun LEE ; Yu Jin HWANG ; Yong Sik KIM
The Korean Journal of Physiology and Pharmacology 2004;8(5):245-251
The effects of fetal mesencephalic cell grafts on the restoration of nigrostriatal dopaminergic function were studied in the intrastriatal 6-hydroxydopamine-lesioned rats. Four weeks after lesioning, transplantation of ventral mesencephalic cells from embryonic day 14 fetuses showed the number of tyrosine hydroxylase (TH) positive cells and fiber outgrowth in the grafted striatum, and significantly ameliorated symptomatic motor behavior of the animals, as determined by apomorphine-induced rotation. Furthermore, in substantia nigra pars compacta (SNc), the numbers of TH cells and fibers were markedly restored. Dopamine content of ipsilateral SNc was close to that of contralateral SNc (91.9 9.8%) in the transplanted animals, while the ratio was approximately 32% in sham-grafted animals. These results indicate that grafted cells restored the activity for the dopaminergic neurons located in SNc, although they were transplanted into striatum. In addition, we showed that the implanted fetal cells expressed high level of glial cell line-derived neurotrophic factor (GDNF), suggesting that the transplanted fetal cells might serve as a dopamine producer and a reservoir of neurotrophic factors. These results may be helpful in consideration of the therapeutic transplantation at early stage of PD.
Animals
;
Dopamine
;
Dopaminergic Neurons
;
Fetus
;
Glial Cell Line-Derived Neurotrophic Factor
;
Nerve Growth Factors
;
Oxidopamine
;
Parkinson Disease
;
Rats*
;
Substantia Nigra
;
Transplantation
;
Transplants*
;
Tyrosine 3-Monooxygenase
2.Effect of Minocycline on Activation of Glia and Nuclear Factor kappa B in an Animal Nerve Injury Model.
Eun Young GU ; Hyung Soo HAN ; Jae Sik PARK
The Korean Journal of Physiology and Pharmacology 2004;8(5):237-243
Glial cells are activated in neuropathy and play a key role in hyperalgesia and allodynia. This study was performed to determine whether minocycline could attenuate heat hyperalgesia and mechanical allodynia, and how glial cell activation and nuclear factor kappa B (NF-kappaB) were regulated by minocycline in a model of chronic constriction of sciatic nerve (CCI). When minocycline (50 mg/kg, oral) was daily administered from 1 day before to 9 days after ligation, heat hyperalgesia and mechanical allodynia were attenuated. Furthermore, when minocycline treatment was initiated 1 or 3 days after ligation, attenuation of the hypersensitive behavior was still robust. However, the effect of attenuation was less when minocycline was started from day 5. In order to elucidate the mechanism of pain attenuation by minocycline, we examined the changes of glia and NF-kappaB, and found that attenuated hyperalgesia and allodynia by minocycline was accompanied by reduced microglial activation. Furthermore, the number of NF-kappaB immunoreactive cells increased after CCI treatment and this increase was attenuated by minocycline. We also observed translocation of NF-kappaB into the nuclei of activated glial cells. These results suggest that minocycline inhibits activation of glial cells and NF-kappaB, thereby attenuating the development of behavioral hypersensitivity to stimuli.
Animals*
;
Constriction
;
Hot Temperature
;
Hyperalgesia
;
Hypersensitivity
;
Ligation
;
Minocycline*
;
Neuroglia*
;
NF-kappa B*
;
Sciatic Nerve
;
Spinal Cord
3.Mechanism of Leptin-Induced Potentiation of Catecholamine Secretion Evoked by Cholinergic Stimulation in the Rat Adrenal Medulla.
Dong Yoon LIM ; Deok Ho CHOI ; Moo Jin KANG
The Korean Journal of Physiology and Pharmacology 2004;8(4):227-235
The aim of the present study was to examine the effect of leptin on CA release from the isolated perfused model of the rat adrenal gland, and to establish its mechanism of action. Leptin (1~100 ng/ml), when perfused into an adrenal vein of the rat adrenal gland for 60 min, enhanced a dose-dependently the secretory responses of CA evoked by ACh (5.32x10 (-3)M), DMPP (10 (-4)M) and McN-A-343 (10 (-4)M), although it alone has weak effect on CA secretion. However, it did not affect the CA secretion evoked by excess K+ (5.6x10 (-2)M). Leptin alone produced a weak secretory response of the CA. Moreover, leptin (10 ng/ml) in to an adrenal vein for 60 min also augmented the CA release evoked by BAY-K-8644, an activator of the dihydropyridine L-type Ca2+ channels, and cyclopiazonic acid, an inhibitor of cytoplasmic Ca2+ ATPase. However, in the presence of U0126 (1micrometer), an inhibitor of mitogen-activated protein kinase (MAPK), leptin no longer enhanced the CA secretion evoked by ACh and DMPP. Furthermore, in the presence of anti-leptin (10 ng/ml), an antagonist of Ob receptor, leptin (10 ng/ml) also no longer potentiated the CA secretory responses evoked by DMPP and Bay-K-8644. Collectively, these experimental results suggest that leptin enhances the CA secretion from the rat adrenal medulla evoked by cholinergic stimulation (both nicotininc and muscarinic receptors), but does not that by membrane depolarization. It seems that this enhanced effect of leptin may be mediated by activation of U0126-sensitive MAPK through the leptin receptors, which is probably relevant to the activation of the dihydropyridine L-type Ca2+ channels located on the rat adrenomedullary chromaffin cells.
(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethylammonium Chloride
;
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
;
Adrenal Glands
;
Adrenal Medulla*
;
Animals
;
Calcium-Transporting ATPases
;
Chromaffin Cells
;
Cytoplasm
;
Dimethylphenylpiperazinium Iodide
;
Leptin
;
Membranes
;
Protein Kinases
;
Rats*
;
Receptors, Leptin
;
Veins
4.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
The Korean Journal of Physiology and Pharmacology 2004;8(4):219-225
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 (EC50; 41.4microliterM and 64.0microliterM, 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 alpha3beta4 nAChR potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons (IC50; 0.53micrometer and 0.22micrometer, respectively). Conversely, alpha- bungarotoxin, alpha-methyllycaconitine, and dihydro-beta-erythroidine, which are known as potent and sensitive blockers for alpha7 or alpha4beta2 nAChRs, below micromolar concentrations showed negligible effect. RT-PCR analysis revealed that alpha3 and beta4 subunits were predominantly expressed in MPG neurons. We suggest that MPG neurons have nAChRs containing alpha3 and beta4 subunits, and that their activation induces fast inward currents, possibly mediating the excitatory synaptic transmission in pelvic autonomic ganglia.
Acetylcholine
;
Animals
;
Dihydro-beta-Erythroidine
;
Ganglia
;
Ganglia, Autonomic
;
Ganglion Cysts*
;
Humans
;
Male*
;
Mecamylamine
;
Negotiating
;
Neurons*
;
Penis
;
Prostate
;
Rats*
;
Receptors, Nicotinic
;
Reverse Transcriptase Polymerase Chain Reaction
;
RNA
;
Synaptic Transmission
;
Urinary Bladder
5.Secondary Fish-Odor Syndrome Can be Acquired by Nitric Oxide-mediated Impairment of Flavin-containing Monooxygenase in Hepatitis B Virus-Infected Patients.
Hyeon Gyu YI ; Jung Nam LEE ; Seung Duk RYU ; Ju Hee KANG ; Young Nam CHA ; Chang Shin PARK
The Korean Journal of Physiology and Pharmacology 2004;8(4):213-218
Primary fish-odor syndrome (FOS) is a genetic disorder caused by defective flavin-containing monooxygenase 3 gene (FMO3) with deficient N-oxidation of trimethylamine (TMA), causing trimethylaminuria (TMAU). By contrast, secondary FOS can be acquired by decreased FMO activities in patients with chronic liver diseases, but the underlying mechanisms are unknown. In the present study, we examined plasma NOx concentrations and viral DNA contents as well as in vivo FMO activities and their correlations in chronic viral hepatitis (CVH) patients. Plasma concentration of NOx was significantly increased by 2.1 fold (56.2+/-26.5 vs. 26.6+/-5.4micrometer, p< 0.01), and it was positively correlated with plasma hepatitis B virus (HBV) DNA contents (r2=0.2838, p=0.0107). Furthermore, the elevated plasma NOx values were inversely and significantly correlated with in vivo FMO activities detected by ranitidine-challenged test (8.3% vs. 20.0%, r2=0.2109, p=0.0315). TMA N-oxidation activities determined in CVH patients without challenge test were also significantly low (73.6% vs. 95.7%, p< 0.05). In conclusion, these results suggested that secondary FOS could be acquired by the endogenously elevated NO in patients with CVH.
DNA
;
DNA, Viral
;
Hepatitis B virus
;
Hepatitis B*
;
Hepatitis*
;
Humans
;
Liver Diseases
;
Nitric Oxide
;
Plasma
;
Ranitidine
6.Identification of a Marker Protein for Cardiac Ischemia and Reperfusion Injury by Two-Dimensional Gel Electrophoresis and Matrix-Assisted Laser Desorption Ionization Mass Spectrometry.
Youngsuk LEE ; Nari KIM ; Hyunju KIM ; Hyun JOO ; Youngnam KIM ; Daehoon JEONG ; Dang Van CUONG ; Euiyong KIM ; Dae Young HUR ; Young Shik PARK ; Yong Geun HONG ; Sangkyung LEE ; Joonyong CHUNG ; Daehyun SEOG ; Jin HAN
The Korean Journal of Physiology and Pharmacology 2004;8(4):207-211
The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.
Blotting, Western
;
Digestion
;
Electrophoresis, Gel, Two-Dimensional*
;
Gels
;
Heart
;
Ischemia*
;
Ischemic Preconditioning
;
Mass Spectrometry*
;
Myoglobin
;
Reperfusion Injury*
;
Reperfusion*
;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
;
Trypsin
7.Detection of Mitochondrial ATP-Sensitive Potassium Channels in Rat Cardiomyocytes.
Dang Van CUONG ; Nari KIM ; Euiyong KIM ; Youngsuk LEE ; Hyunju KIM ; Sunghyun KANG ; Dae Young HUR ; Hyun JOO ; Young Shik PARK ; Yong Geun HONG ; Sangkyung LEE ; Joonyong CHUNG ; Daehyun SEOG ; Jin HAN
The Korean Journal of Physiology and Pharmacology 2004;8(4):201-206
Mitochondrial ATP-sensitive potassium (mitoKATP) channels play a role in early and late ischemic preconditioning. Nevertheless, the subunit composition of mitoKATP channels remains unclear. In this study, we investigated the subunit composition of mitoKATP channels in mitochondria isolated from rat cardiac myocytes. Mitochondria were visualized using the red fluorescence probe, Mitrotracker Red, while mitoKATP channels were visualized using the green fluorescence probe, glibenclamide-BODIPY. The immunofluorescence confocal microscopy revealed the presence of Kir6.1, Kir6.2 and SUR2 present in the cardiac mitochondria. Western blot analysis was carried to further investigate the nature of mitoKATP channels. For SUR proteins, a 140-kDa immunoreactive band that corresponded to SUR2, but no SUR1 was detected. For Kir6.2, three bands (~4, ~6, and ~0 kDa) were detected, and a specific ~6-kDa immunoreactive band corresponding to Kir6.1 was also observed. These observations suggest that the subunits of mitoKATP channels in rat myocytes include Kir6.1, Kir6.2, and a SUR2-related sulfonylurea-binding protein.
Animals
;
Blotting, Western
;
Fluorescence
;
Fluorescent Antibody Technique
;
Ischemic Preconditioning
;
KATP Channels*
;
Microscopy, Confocal
;
Mitochondria
;
Muscle Cells
;
Myocytes, Cardiac*
;
Potassium
;
Rats*
8.The Effect of Ghrelin on Ca2+ Concentration in Thyroid FRTL-5 Cells.
Byung Joo KIM ; Young Joo PARK ; Do Joon PARK ; Insuk SO ; Ki Whan KIM
The Korean Journal of Physiology and Pharmacology 2004;8(4):195-200
Ghrelin is a newly discovered peptide, which is released from the stomach and neurons in the hypothalamic arcuate nucleus (ARC), and potently stimulates growth hormone release and food intake. In the present study, we investigated the effect of ghrelin on [Ca2+]i in thyroid FRTL-5 cells. Ghrelin (5 nM) increased [Ca2+]i and TSH (1 unit/l) had an additive effect on [Ca2+]i when extracellular normal solution was 1.1mM Ca2+ containing Coon's modified Ham's F12 medium. When Ca2+-free medium containing 2 mM EGTA replaced the above normal solution, ghrelin also induced a similar rise in [Ca2+]i. In the middle of [Ca2+]i increment by ghrelin, nifedipine (1 micrometer), nickel (100micrometer) and La3+ (100micrometer) had no effect on [Ca2+]i. After endoplasmic reticulum was depleted by cyclopiazonic acid (CPA; 10micrometer), ghrelin caused no visible change on [Ca2+]i in Ca2+-free/2 mM EGTA solution. These results suggest that ghrelin can increase [Ca2+]i through endoplasmic reticulum in thyroid FRTL-5 cells.
Arcuate Nucleus
;
Eating
;
Egtazic Acid
;
Endoplasmic Reticulum
;
Ghrelin*
;
Growth Hormone
;
Neurons
;
Nickel
;
Nifedipine
;
Stomach
;
Thyroid Gland*
9.Effect of Mild Hypothermia on the Mitogen Activated Protein Kinases in Experimental Stroke.
The Korean Journal of Physiology and Pharmacology 2004;8(4):187-194
Middle cerebral artery occlusion (MCAO) results in cell death by activation of complex signal pathways for cell death and survival. Hypothermia is a robust neuroprotectant, and its effect has often been attributed to various mechanisms, but it is not yet clear. Upstream from the cell death promoters and executioners are several enzymes that may activate several transcription factors involved in cell death and survival. In this study, we immunohistochemically examined the phosphorylation of mitogen- activated protein kinase, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 kinase during early period of the ischemic injury, following 2 hours (h) of transient MCAO. Increased phosphorylation of ERK and p38 was observed in the vessels at 3 h, neuron-like cells at 6 and 12 h and glia-like cells at 12 h. Activation of JNK was not remarkable, and a few cells showed active JNK following ischemia. Phosphorylation of Elk-1, a transcription factor, was reduced by ischemic insult. Hypothermia attenuated the activation of ERK, p38 and JNK, and inhibited reduction of Elk-1. These data suggest that signals via different MAPK family members converge on the cell damage process and hypothermia protects the brain by interfering with these pathways.
Brain
;
Cell Death
;
Humans
;
Hypothermia*
;
Infarction, Middle Cerebral Artery
;
Ischemia
;
JNK Mitogen-Activated Protein Kinases
;
Mitogen-Activated Protein Kinases*
;
Phosphorylation
;
Phosphotransferases
;
Protein Kinases
;
Signal Transduction
;
Stroke*
;
Transcription Factors
10.Activation of Vestibular Neurons Projecting to Autonomic Brain Stem Nuclei Following Acute Hypotension in Rats.
Dong Ok CHOI ; Chon Il YON ; Myoung Ae CHOI ; Byung Rim PARK ; Min Sun KIM
The Korean Journal of Physiology and Pharmacology 2004;8(4):181-185
Extracellular regulated protein kinase1/2 (pERK1/2) is one of the major regulatory factors for transcription of the c-fos oncogene in neurons. The purpose of this study was to evaluate the expression of phosphorylated ERK1/2 within the vestibular nuclei (VN) of rats following acute arterial hypotension. Following the acute arterial hypotension induced by rapid hemorrhage, a significant number of pERK1/2-immunoreactive neurons appeared bilaterally in the caudal aspect of the medial and inferior VN. No labeling of pERK1/2 was observed in the lateral VN. The peak expression of pERK1/2 in these nuclei occurred within 5 min after hemorrhage. However, in bilaterally labyrinthectomized rats, the appearance of pERK1/2-immunoreactive neurons was eliminated in the VN. Western blot confirmed the effect of bilateral labyrinthectomy on pERK1/2 protein expression in the medial vestibular nucleus 5 min after hemorrhage. These results suggest that, following acute hypotension, afferent signals from the peripheral vestibular receptors are required for activation of ERK 1/2 in the VN.
Animals
;
Blotting, Western
;
Brain Stem*
;
Brain*
;
Hemorrhage
;
Hypotension*
;
Neurons*
;
Oncogenes
;
Rats*
;
Vestibular Nuclei