Biomedical subjects

Tae-Cheon Kang

Publications and source records attributed to Tae-Cheon Kang.

At least 91 records · Page 5Linked to original sources

PubMed2004

Expression and changes of endogenous insulin-like growth factor-1 in neurons and glia in the gerbil hippocampus and dentate gyrus after ischemic insult.

In the present study, we focused upon expression and changes of endogenous insulin-like growth factor-1 (IGF-1) in the hippocampus of the Mongolian gerbil after ischemic insult. In sham-operated animals, IGF-1 immunoreactivity was absent from the hippocampus. IGF-1-immunoreactive (IR) neurons were detected at 12 h and 1 day after ischemic insult. In the hippocampal CA1 area, the IGF-IR neurons were non-pyramidal cells (GABAergic neurons). In the hippocampal CA2/3 areas, the IGF-1-IR neurons were pyramidal and non-pyramidal cells, and in the dentate gyrus the IGF-1-IR neurons were hilar neurons. Four days after ischemia-reperfusion, IGF-1 immunoreactivity disappeared from neurons, and significantly increased in astrocytes and microglia. These results suggest that the induction of IGF-1 in the CA1 area during the early stage (12-24 h after ischemic insult) is associated with the relative vulnerabilities of pyramidal glutamatergic neurons and non-pyramidal GABAergic neurons. The later increase (4 days after ischemic insult) of IGF-1 expression and protein content was found to promote the activities of astrocytes and microglia. These increases of IGF-1 in astrocytes and in microglia are associated with mechanisms that compensate for the effects of delayed neuronal death.

Animals↗

PubMed2004

Altered Na+-K+ ATPase immunoreactivity within GABAergic neurons in the gerbil hippocampal complex induced by spontaneous seizure and vigabatrin treatment.

In the present study, the expression of Na(+)-K(+) ATPase in the gerbil hippocampus associated with various sequelae of spontaneous seizures were investigated in order to identify the roles of Na(+)-K(+) ATPase in the epileptogenesis and the recovery mechanisms in these animals. The population of Na(+)-K(+) ATPase immunoreactive neurons and Na(+)-K(+) ATPase immunodensity were significantly lower in the pre-seizure group of SS gerbils than those in SR gerbils. At 30-min postictal, the Na(+)-K(+) ATPase immunoreactivity was significantly elevated in the hippocampal complex. At 3-h postictal, the Na(+)-K(+) ATPase immunoreactivity in the hippocampus was declined, as compared to the 30-min postictal. At 12h after seizure on-set, Na(+)-K(+) ATPase expression was re-enhanced in the all regions of the hippocampal complex including the dentate hilus. Following administration of vigabatrin Na(+)-K(+) ATPase expression was also increased. The present data suggest that altered Na(+)-K(+) ATPase expression may contribute the regulation of the seizure activity in this animal.

Animals↗

PubMed2004

Altered GABAB receptor immunoreactivity in the gerbil hippocampus induced by baclofen and phaclofen, not seizure activity.

The present study was performed to determine whether the effects induced by GABA(B) receptor-acting drugs would be related with the alteration in GABA(B) receptor expression in the hippocampus using Mongolian gerbil, a genetic epilepsy model. The distribution patterns of both GABA(B) receptor 1A/B and GABA(B)receptor 2 immunoreactivities were similarly detected in the hippocampi of normal and seizure-prone gerbils. Following baclofen (GABA(B) receptor agonist) or phaclofen (GABA(B) receptor antagonist) treatment, GABA(B) receptor immunoreactivities were decreased or increased by dose-dependent manners, respectively. Vigabatrin (GABA transaminase inhibitor) or 3-mercaptopropionic acid (GAD inhibitor) treatment did not affect GABA(B) receptor expressions. These findings suggest that GABA(B) receptor expression in the gerbil hippocampus may be altered by baclofen or phaclofen treatment.

Animals↗

PubMed2004

Changes in the expression of calbindin D-28k in the gerbil hippocampus following seizure.

Previous studies have reported that calbindin D-28k (CB), a calcium-binding protein, containing neurons in the hippocampus play an important role in hippocampal excitability in epilepsy, because CB modulates the free calcium ion during seizure. Hence, in the present study, we investigated changes of CB expression in the hippocampus and its association in the Mongolian gerbil to identify roles of CB in epileptogenesis. CB immunoreactivity in the hippocampus was significantly lower in the pre-seizure group of seizure sensitive (SS) gerbils as compared with those seen in the seizure resistant (SR) gerbils. The distribution of CB immunoreactivity in the hippocampus showed significant difference after seizure on-set in SS gerbils. CB immunoreactivity in the hippocampal CA1, CA2 areas, and subiculum was lowest at 3h after seizure on-set; thereafter, the immunoreactivity became to increase to 12h after seizure on-set. Mossy fibers, Schaffer collaterals and dentate granule cells showed the highest CB immunoreactivity at 3h after seizure on-set; thereafter, the immunoreactivity became to decrease. In the case of the intrinsic and output connections of the hippocampus, a rapid decrease of CB serves an inhibitory function, which regulates the seizure activity and output signals from the hippocampus.

Animals↗

PubMed2004

Vigabatrin inhibits pyridoxine-5'-phosphate oxidase, not pyridoxal kinase in the hippocampus of seizure prone gerbils.

To identify the effects of vigabatrin (VGB) on the metabolism of pyridoxal 5'-phosphate (PLP) in the seizure prone gerbil hippocampus, we conducted a chronological and comparative analysis of pyridoxal kinase (PLK) and pyridoxine-5'-phosphate oxidase (PNP oxidase) expression. In the VGB treated animals, PNP oxidase immunoreactivity was reduced, although the distribution and immunodensity of PLK were unaltered, as compared with control animals. In a Western blot study, the densities of PNP oxidase immunoreactivities in VGB treated animals were found to have decreased significantly. However, no differences in PLK immunoreactive bands were observed in controls or in VGB treated animals. By enzyme activity assay, and in contrast to PLK, the specific activity of PNP oxidase in the VGB treated gerbils was significantly reduced. In conclusion, the present data presents a piece of in vivo evidence that supports the anti-epileptic effects mediated by pyridoxamine-5'-phosphate (PMP) metabolism, and which may be helpful in the development of an anti-epileptic drug.

Animals↗

PubMed2004

Soy isoflavones improve spatial delayed matching-to-place performance and reduce cholinergic neuron loss in elderly male rats.

To investigate the protective activity of soy isoflavones on neurons, the effects of isoflavones on cholinergic enzyme activity, immunoreactivities of cholinergic enzyme, and delayed matching-to-place (DMP) performance were measured in normal elderly rats. Male Sprague-Dawley rats (n = 48; 10 mo old) were assigned to 3 groups: CD (control diet), ISO 0.3 (0.3 g/kg soy isoflavones diet), and ISO 1.2 (1.2 g/kg soy isoflavones diet). After 16 wk of consuming these diets, choline acetyltransferase (ChAT) activity in the ISO 0.3 group was greater in cortex and basal forebrain (BF; P < 0.05) than in controls. In BF, ChAT activity was also significantly greater in the ISO 1.2 group than in control rats. Acetylcholine esterase (AChE) activity in the ISO 0.3 group was significantly inhibited in cortex, BF, and hippocampus and in the ISO 1.2 group in cortex and hippocampus. Choline acetyltransferase immunoreactivity (ChAT-IR) in the ISO 1.2 group was significantly greater than in controls in the medial septum area. ChAT-IR in the ISO 0.3 and ISO 1.2 groups was significantly higher than in the CD group in the hippocampus CA1 area. Spatial DMP performance by the ISO 0.3 group showed significantly shorter swimming time than by the CD group. These findings show that soy isoflavones can influence the brain cholinergic system and reduce age-related neuron loss and cognition decline in male rats.

Aging↗

PubMed2004

Genomic organization, tissue distribution and deletion mutation of human pyridoxine 5'-phosphate oxidase.

We used a combined computer and biochemical approach to characterize human pyridoxine 5'-phosphate oxidase (PNPO). The human PNPO gene is composed of seven exons and six introns, and spans approximately 8 kb. All exon/intron junctions contain the gt/ag consensus splicing site. The absence of TATA-like sequences, the presence of Sp1-binding sites and more importantly, the presence of CpG islands in the regulatory region of the PNPO gene are characteristic features of housekeeping genes. Northern blot analyses showed two species of poly(A)(+) RNA of approximately 2.4 and approximately 3.4 kb at identical intensity, whereas Western blot analysis showed that no protein isoform exists in any of the tissues examined. PCR-based analysis led to the idea that two messages are transcribed from a single copy gene, and that the size difference is due to differential usage of the polyadenylation signal. The major sites of PNPO expression are liver, skeletal muscle and kidneys while a very weak signal was detected in lung. The mRNA master dot-blot for multiple human tissues provided a complete map of the tissue distribution not only for PNPO but also for pyridoxal kinase and pyridoxal phosphatase. The data indicate that mRNA expression of all three enzymes essential for vitamin B(6) metabolism is ubiquitous but is highly regulated at the level of transcription in a tissue-specific manner. In addition, human brain PNPO cDNA was expressed in Escherichia coli, and the roles of both the N- and C-terminal regions were studied by creating sequential truncation mutants. Our results showed that deletion of the N-terminal 56 residues affects neither the binding of coenzyme nor catalytic activity.

Alternative Splicing↗

PubMed2004

Brain succinic semialdehyde dehydrogenase. Reactions of sulfhydryl residues connected with catalytic activity.

Incubation of an NAD+-dependent succinic semialdehyde dehydrogenase from bovine brain with 4-dimethylaminoazobenzene-4-iodoacetamide (DABIA) resulted in a time-dependent loss of enzymatic activity. This inactivation followed pseudo first-order kinetics with a second-order rate constant of 168 m(-1).min(-1). The spectrum of DABIA-labeled enzyme showed a characteristic peak of the DABIA alkylated sulfhydryl group chromophore at 436 nm, which was absent from the spectrum of the native enzyme. A linear relationship was observed between DABIA binding and the loss of enzyme activity, which extrapolates to a stoichiometry of 8.0 mol DABIA derivatives per mol enzyme tetramer. This inactivation was prevented by preincubating the enzyme with substrate, succinic semialdehyde, but not by preincubating with coenzyme NAD+. After tryptic digestion of the enzyme modified with DABIA, two peptides absorbing at 436 nm were isolated by reverse-phase HPLC. The amino acid sequences of the DABIA-labeled peptides were VCSNQFLVQR and EVGEAICTDPLVSK, respectively. These sites are identical to the putative active site sequences of other brain succinic semialdehyde dehydrogenases. These results suggest that the catalytic function of succinic semialdehyde dehydrogenase is inhibited by the specific binding of DABIA to a cysteine residue at or near its active site.

Aldehyde Oxidoreductases↗

PubMed2003

Elevated P/Q type (alpha1A) and L2 type (alpha1D) Purkinje cell voltage-gated calcium channels in the cerebella of seizure prone gerbils.

Differences in expression of N-methyl-D-aspartate (NMDA) receptor and voltage gated Ca2+ channels (VGCC) in the gerbil cerebellum were investigated to identify routes of Ca2+ influx that may be involved in Purkinje cell damage. Immunodensities of NR1 and NR2A/B were the same in seizure resistant (SR) and seizure sensitive (SS) gerbils. However, both P/Q type (alpha1A) and L2 type (alpha1D) VGCC levels were higher in the Purkinje cells of SS gerbils than in those of SR gerbils, whereas N type (alpha1B) and L1 type (alpha1C) VGCC levels were similar in the two groups. Our findings suggest that increases in P/Q type (alpha1A) and L2 type (alpha1D) VGCC are implicated in the degeneration of Purkinje cells in SS gerbils.

Animals↗

PubMed2003

Age-related changes of parvalbumin immunoreactive neurons in the rat main olfactory bulb.

Parvalbumin (PV) is found in the olfactory system, including the main olfactory bulb, and is thought to be one of the neuroactive substances in olfaction. Changes in PV immunoreactivity in the olfactory system during aging have not been examined. We investigated such changes in the main olfactory bulb (MOB) of the rat at postnatal month 1 (PM 1), PM 3, PM 6, PM 12 and PM 24. PV-IR neurons were almost completely restricted to the external plexiform layer. At PM 1 there were only a few PV-IR neurons; at PM 3, the number of PV-IR neurons was at its greatest but they were not well developed morphologically. At PM 6, the number of PV-IR neurons was similar to that at PM 3 and they had satellite somata with well-developed processes with many varicosities. By PM 12 the number of neurons and processes had declined, and by PM 24, they had fallen even further and the remaining processes had lost most of their varicosities. We conclude that age-related degeneration of PV-IR neurons in the MOB may reduce calcium buffering and affect olfactory function in senile species.

Aging↗

PubMed2003

Changes of glial Na+-K+ ATPase (alpha 1 subunit) immunoreactivity in the gerbil hippocampus after transient forebrain ischemia.

In the present study to evaluate the effects of ischemia on sodium-potassium adenosine triphosphatase (Na(+)-K+ ATPase) alpha1 subunit (alpha6F) expression in the glia, the immunodensities of both Na(+)-K+ ATPase and the glial fibrillary acidic protein in the hippocampus were measured and analyzed. In the sham hippocampus, alpha6F immunoreactivity was mainly observed in the both the molecular layer and the polymorphic layer of dentate gyrus. At 30 min after ischemic insult, the alpha6F immunoreactivity was markedly decreased in the molecular layer of the dentate gyrus, in contrast to the appearance of this immunoreactivity in the hilar neurons. Up to 12 h after ischemic insult, the alpha6F immunoreactivity was re-enhanced in the molecular layer of dentate gyrus. In addition, the alpha6F immunoreactivity appeared slightly in the glial components in the hippocampal region. Four days after ischemia-reperfusion, the intensity of alpha6F immunoreactivity in the glial cells was highest. At this time point, strong alpha6F immunoreactivity was colocalized with GFAP immunoreactivity in the strata radiatum of the CA1 and the molecular layer of the dentate gyrus. These results suggest that the enhancement of alpha6F immunoreactivity may be a compensatory response to regulate the ion homeostasis in the brain. In addition, the maintenance of Na(+)-K+ ATPase activity in the astrocytes may explain the resistant characteristics of these cells to ischemic insults.

Animals↗

PubMed2003

The somatostatin receptors in the normal and epileptic hippocampus of the gerbil: subtype-specific localization and its alteration.

We investigated the distribution of somatostatin receptors (SSTs) in the hippocampi of SR (seizure-resistant) and SS (seizure-sensitive) gerbils in order to characterize the alterations in SST expressions induced by seizure activity. SST2A immunodensity in the hippocampus of SS gerbils was lower than that of SR gerbils, though its localization in the hippocampus was similar in both SR and SS gerbils. SST3 immunodensity in the hippocampus of SS gerbils was lower than in SR gerbils. In SR gerbils, strong SST4 immunoreactivity was detected in the dentate gyrus and in the CA3 region, in contrast little immunoreactivity was detected in these regions in SS gerbils. In SR and SS gerbils, the strong SST5 immunoreactivity in the hippocampus was also detected in the stratum oriens of the CA2-3 regions and the septal area of CA1 region. However, SST5 immunodensity in the stratum radiatum in SS gerbils was lower than in SR gerbils. These results are the first comprehensive description of the distribution of SSTs in the normal and epileptic hippocampus of gerbils, and suggest that these alterations in the hippocampus of the SS gerbil may be related with a regulatory mechanism for seizure activity in these seizure prone animals.

Animals↗

PubMed2003

Human pyridoxal phosphatase. Molecular cloning, functional expression, and tissue distribution.

Pyridoxal phosphatase catalyzes the dephosphorylation of pyridoxal 5'-phosphate (PLP) and pyridoxine 5'-phosphate. A human brain cDNA clone was identified to the PLP phosphatase on the basis of peptide sequences obtained previously. The cDNA predicts a 296-amino acid protein with a calculated Mr of 31698. The open reading frame is encoded by two exons located on human chromosome 22q12.3, and the exon-intron junction contains the GT/AG consensus splice site. In addition, a full-length mouse PLP phosphatase cDNA of 1978 bp was also isolated. Mouse enzyme encodes a protein of 292 amino acids with Mr of 31512, and it is localized on chromosome 15.E1. Human and mouse PLP phosphatase share 93% identity in protein sequence. A BLAST search revealed the existence of putative proteins in organism ranging from bacteria to mammals. Catalytically active human PLP phosphatase was expressed in Escherichia coli, and characteristics of the recombinant enzyme were similar to those of erythrocyte enzyme. The recombinant enzyme displayed Km and kcat values for pyridoxal of 2.5 microM and 1.52 s(-1), respectively. Human PLP phosphatase mRNA is differentially expressed in a tissue-specific manner. A single mRNA transcript of 2.1 kb was detected in all human tissues examined and was highly abundant in the brain. Obtaining the molecular properties for the human PLP phosphatase may provide new direction for investigating metabolic pathway involving vitamin B6.

Amino Acid Sequence↗

PubMed2003

Isolation and identification of an antioxidant enzyme catalase stimulatory compound from Garnoderma lucidum.

Antioxidant enzymes are scavenger reactive-oxygen intermediates and are involved in many cellular defense systems. We previously reported that a crude extract of Garnoderma lucidum, a medicinally potent mushroom, profoundly increased the catalase gene expression and enzyme activities in mouse livers (Park et al., J. Biochem. Mol. Biol. 34. 144-149, 2001). In this study, we elucidated the detailed mechanism whereby G. lucidum stimulates the catalase activity and expression. The major active fraction was isolated from G. lucidum and methyl linoleate was considered the most major component of the fraction. In order to determine whether methyl linoleate increases mRNA and protein synthesis of catalase, Northern and Western blot analyses were performed in vivo with methyl linoleate-treated mouse liver homogenate after feeding methyl linoleate to the mice. Northern and Western blot analyses of the crude liver homogenates in the mice that were administered methyl linoleate revealed that the expression catalase was significantly increased when compared to the untreated controls. In addition, the catalase protein levels and enzymatic activities increased in the mouse liver homogenates. These results suggest that methyl linoleate that is produced by G. lucidum stimulates the catalase expression at the transcription level.

Agaricales↗

PubMed2003

Human glutamate dehydrogenase is immunologically distinct from other mammalian orthologues.

Five monoclonal antibodies (mAbs) that recognize human glutamate dehydrogenase (GDH) have been selected and designated as monoclonal antibodies hGDH60-6, hGDH60-8, hGDH63-10, hGDH63-11, and hGDH91-14. A total of five mAbs recognizing different epitopes of the enzyme were obtained, two of which inhibited human GDH activity. When total proteins of human homogenate separated by SDS- PAGE, were probed with mAbs, a single reactive protein band of 55 kDa, which co-migrated with purified recombinant human GDH was detected. When the purified GDH was incubated with each of the mAbs, its enzyme activity was inhibited by up to 58%. Epitope mapping analysis identified, two subgroups of mAbs recognizing different peptide fragments. Using the individual anti-GDH antibodies as probes, the cross reactivities of brain GDH obtained from human and other animal brain tissues were investigated. For the human and animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 55 kDa when hGHD60-6, hGHD60-8, or hGHD91-14 mAbs were used as probes. However, the anti-human GDH mAbs immunoreactive to bands on Western blots reacted differently on the immunoblots of the other animal brains tested, i.e., the two monoclonal antibodies hGDH63-10 and hGDH63-11 only produced positive results for human. These results suggest that human brain GDH is immunologically distinct from those of other mammalian brains. Thorough characterization of these anti-human GDH mAbs could provide potentially valuable tool as immunodiagnostic reagents for the detection, identification and characterization of the various neurological diseases related to the GDH enzyme.

Animals↗

PubMed2003

P2X2 and P2X4 receptor expression is regulated by a GABA(A) receptor-mediated mechanism in the gerbil hippocampus.

Fast responses to extracellular ATP are mediated by the activation of P2X receptors. Native and cloned P2X receptors are permeable to monovalent cations such as Na+ and K+ as well as divalent cations such as Ca2+. However, altered P2X receptor expression has not been definitively determined under pathological conditions, particularly in epilepsy. Here we show that, in the seizure-sensitive (SS) gerbil hippocampus, a recognized genetic epilepsy model, the expressions of both P2X2 and P2X4 receptors are markedly decreased as compared with that in the seizure-resistant (SR) gerbil. These alterations are closely related to changes in gamma-aminobutyric acid (GABA) concentrations induced by vigabatrin (VGB) or 3-mercaptopropionic acid (3-MPA) treatment. Furthermore, the regulation of both P2X receptor expression in the gerbil hippocampus was mediated by the GABA(A) receptor, not GABA(B). These results suggest that the GABA(A) receptor-mediated modulation of P2X receptor expression may play an important role in the regulation of neuronal excitability.

3-Mercaptopropionic Acid↗

PubMed2003

Presynaptic gamma-aminobutyric acid type B receptor-mediated regulation of vesicular gamma-aminobutyric acid transporter expression in the gerbil hippocampus.

Changes in vesicular gamma-aminobutyric acid (GABA) transporter (VGAT) expression in the gerbil hippocampus after treatment with baclofen (GABA(B) receptor agonist) or phaclofen (GABA(B) receptor antagonist) were investigated to identify the GABA(B) receptor-mediated regulation of VGAT expression. In the baclofen-treated seizure-resistant gerbils, VGAT expression was significantly reduced, as compared with the control animals, thus the VGAT immunoreactive pattern in these gerbils was similar to that in control seizure-sensitive (SS) gerbils. In the phaclofen-treated SS gerbils, VGAT expression was dramatically elevated, compared to SS gerbil controls. Our findings demonstrated that GABA(B) receptor-mediated regulation of VGAT expression may be another mechanisms for presynaptic GABA release, which is accompanied by a reduction in Ca(2+) conductance by the inhibition of voltage-gated Ca(2+) channels.

Animals↗

PubMed2003

Elevation of Na+-K+ ATPase immunoreactivity in GABAergic neurons in gerbil CA1 region following transient forebrain ischemia.

In a previous study, we suggested that GABAergic neurons might be resistant to ischemic insult, because of the maintenance of the GABA shunt, which is one of the ATP synthetic pathways in neurons. In the present study, we identified Na(+)-K(+) ATPase immunoreactivity in the gerbil hippocampus in order to determine whether changes in Na(+)-K(+) ATPase immunoreactivity correlate with GABA shunt following ischemic insult. At 12 h after ischemia-reperfusion, Na(+)-K(+) ATPase immunoreactivity accumulated in some neurons in the CA1 region. However, the protein content of Na(+)-K(+) ATPase was not altered. Interestingly, the density of Na(+)-K(+) ATPase immunoreactivity in neurons and the protein content in the CA1 region was intensified in the 24 h post-ischemic group. As a result of double immunofluorescence study, Na(+)-K(+) ATPase immunoreactive neurons were identified with GABAergic neurons. Therefore, our findings suggest that the increase of Na(+)-K(+) ATPase in GABAergic neurons may be able to explain the resistance of these cells to ischemic insult, and support our previous hypothesis that GABA may play an important role as a metabolite in the survival of GABAergic neurons after ischemic insult.

Animals↗