Biomedical subjects

Moo Ho Won

Publications and source records attributed to Moo Ho Won.

At least 55 records · Page 3Linked to original sources

PubMed2005

Antioxidant-like protein 1 is altered in non-pyramidal cells and expressed in astrocytes in the gerbil hippocampal CA1 region after transient forebrain ischemia.

In the present study, we observed chronological changes of antioxidant-like protein 1 (AOP-1) in the gerbil hippocampal CA1 region after 5 min of transient forebrain ischemia using immunohistochemistry and western blot. AOP-1 was significantly altered in the CA1 region after transient ischemia. In the sham-operated group, AOP-1 immunoreactivity was detected in pyramidal and non-pyramidal cells of the CA1 region. At 30 min after ischemic insult, AOP-1 immunoreactivity and protein level was decreased in the CA1 region. At 12 h after ischemic insult, AOP-1 immunoreactivity and protein level was highest in this region. At this time, after ischemia, AOP-1 immunoreactivity in non-pyramidal cells was high compared to the sham-operated group. Based on double immunofluorescence study, AOP-1-immunoreactive neurons were identified as GABAergic, which were stained with GAD or parvalbumin. Thereafter, AOP-1 immunoreactivity and protein levels were decreased time-dependently. From 4 days after ischemic insult, AOP 1 immunoreactivity was generally expressed in astrocytes. Five days after ischemic insult, AOP-1 immunoreactivity and protein level was increased again to 1.4 folds compared to that of the sham-operated group. In brief, AOP-1 immunoreactivity was increased in GABAergic non-pyramidal cells in the hippocampal CA1 region at early time after ischemic insult and was expressed in astrocytes at late time after ischemia. This result suggests that AOP-1 may be important role in homeostasis of GABAergic neurons because these neurons are resistant to ischemic damage.

Animals↗

PubMed2005

Age-dependent changes of pyridoxal phosphate synthesizing enzymes immunoreactivities and activities in the gerbil hippocampal CA1 region.

In the present study, age-related changes of pyridoxal 5'-phosphate (PLP) synthesizing enzymes, pyridoxal kinase (PLK) and pyridoxine 5'-phosphate oxidase (PNPO), their protein contents and activities were examined in the gerbil hippocampus proper. Significant age-dependent changes in PLK and PNPO immunoreactivities were found in the CA1 region, but not in the CA2/3 region. In the postnatal month 1 (PM 1) group, PLK and PNPO immunoreactivities were detected mainly in the stratum pyramidale of the CA1 region. PLK and PNPO immunoreactivities and their protein contents were highest in the PM 6 group, showing that many CA1 pyramidal cells had strong PLK and PNPO immunoreactivities. Thereafter, PLK and PNPO immunoreactivities started to decrease and were very low at PM 24. Alterations in the change patterns in protein contents and total activities of PLK and PNPO corresponded to the immunohistochemical data, but their specific activities were not altered in any experimental group. Based on double immunofluorescence study, PLK and PNPO immunoreactive cells in the strata oriens and radiatum were identified as GABAergic cells. Therefore, decreases of PLK and PNPO in the hippocampal CA1 region of aged brains may be involved in aging processes related with gamma-aminobutyric acid (GABA) function.

Age Factors↗

PubMed2005

Bilateral enhancement of excitation via up-regulation of vesicular glutamate transporter subtype 1, not subtype 2, immunoreactivity in the unilateral hypoxic epilepsy model.

In the present study, the change of vesicular glutamate transporter (VGLUT) immunoreactivity on long-term impaired excitability in the hippocampus after recovery from unilateral hypoxic-ischemic insult was investigated in order to extend our understanding of the mechanism of epileptogenesis using unilateral hypoxic epilepsy models. Both the lesioned (submitted to ischemia) and the unlesioned hippocampi exhibited the frequent occurrence of interictal spikes and occasionally the sustained ictal discharges. However, paired-pulse inhibition was significantly reduced in the unlesioned dentate gyrus, not in the lesioned dentate gyrus. VGLUT1 immunoreactivity was significantly elevated in both hippocampi following hypoxic ischemia, although VGLUT2 immunodensity was unaltered. These findings suggest that the enhancement of VGLUT1 immunoreactivity in both hippocampi after unilateral hypoxic ischemia may contribute to the hyperexcitability, which may play an important role in the epileptogenesis (presumably accompanied by altered inhibitory transmission) after neurodegeneration.

Animals↗

PubMed2005

Transient ischemia-induced changes of neurofilament 200 kDa immunoreactivity and protein content in the main olfactory bulb in gerbils.

This study was carried out to investigate alterations of neurofilament 200 kDa (NF-200) and its polyphosphorylation form (RT97) immunoreactivity and protein content in the main olfactory bulb (MOB) after 5 min of transient forebrain ischemia in gerbils. In the sham-operated group, weak NF-200 immunoreactivity was detectable in a few somata of mitral cells, which projected weak NF-200-immunoreactive processes to the external plexiform layer (EPL). At 1-5 days after ischemia, strong NF-200 and RT97 immunoreactivity was shown by the mitral cell processes; however, somata of mitral cells did not show NF-200 immunoreactivity. At this time point, strong NF-200-immunoreactive mitral cell processes ran to the EPL and glomerular layer (GL). Thereafter, NF-200 and RT97 immunoreactivity was decreased up to 30 days after ischemia. In the 15 days post-ischemic group, the distribution pattern of NF-200 and RT97 immunoreactivity was slightly lower than that in the 1-5 days post-ischemic groups. In the 30 days post-ischemic group, moderate NF-200 and RT97 immunoreactivity was found in the mitral cells processes, but the immunoreactivity in the EPL and GL nearly disappeared. A Western blot study showed a pattern of NF-200 and RT97 expression at all post-ischemic time points similar to that of immunohistochemistry after ischemia. This result indicates that NF-200 and RT97 accumulates in injured mitral cell processes a few days after transient ischemia, which suggests that the axonal transport in the MOB may be disturbed during this period after transient ischemia.

Animals↗

PubMed2005

Expression and changes of Ca2+-ATPase in neurons and astrocytes in the gerbil hippocampus after transient forebrain ischemia.

Ca2+-ATPase is one of the most powerful modulators of intracellular calcium levels. In this study, we focused on chronological changes in the immunoreactivity and protein levels of Ca2+-ATPase in the hippocampus after 5 min of transient forebrain ischemia. Ca2+-ATPase immunoreactivity was significantly altered in the hippocampal CA1 region and in the dentate gyrus, but not in the CA2/3 region after ischemic insult. In the sham-operated group, Ca2+-ATPase immunoreactivity was detected in the hippocampus. Ca2+-ATPase immunoreactivity in the CA1 region and in the dentate gyrus, and its protein levels peaked 3 h after ischemic insult. At this time, CA1 pyramidal cells and dentate polymorphic cells showed strong Ca2+-ATPase immunoreactivity. Thereafter, Ca2+-ATPase immunoreactivity reduced in the CA1 region and in the dentate gyrus. One day after ischemic insult, Ca2+-ATPase immunoreactivity was observed in some CA1 non-pyramidal cells, and 4 days after ischemic insult, Ca2+-ATPase immunoreactivity was detected in astrocytes throughout the CA1 region, but Ca2+-ATPase immunoreactivity in the dentate gyrus had nearly disappeared. Our results suggest that Ca2+-ATPase changes may be associated with a response to ischemic damage in hippocampal CA1 pyramidal cells, and that increased Ca2+-ATPase immunoreactivity in the reactive astrocytes may be associated with the maintenance of intracellular calcium levels.

Animals↗

PubMed2005

TNF-related activation-induced cytokine enhances leukocyte adhesiveness: induction of ICAM-1 and VCAM-1 via TNF receptor-associated factor and protein kinase C-dependent NF-kappaB activation in endothelial cells.

Inflammation is a basic pathological mechanism leading to a variety of vascular diseases. The inflammatory reaction involves complex interactions between both circulating and resident leukocytes and the vascular endothelium. In this study, we report evidence for a novel action of TNF-related activation-induced cytokine (TRANCE) as an inflammatory mediator and its underlying signaling mechanism in the vascular wall. TRANCE significantly increased endothelial-leukocyte cell interactions, and this effect was associated with increased expression of the cell adhesion molecules, ICAM-1 and VCAM-1, on the endothelial cells. RT-PCR analysis and promoter assays revealed that expression of these cell adhesion molecules was transcriptionally regulated mainly by activation of the inflammatory transcription factor, NF-kappaB. TRANCE induced IkappaB-alpha phosphorylation and NF-kappaB activation via a cascade of reactions involving the TNFR-associated factors, phospholipase C, PI3K, and protein kinase C (PKC-alpha and PKC-zeta). It also led to the production of reactive oxygen species via PKC- and PI3K-dependent activation of NADPH oxidase in the endothelial cells, and antioxidants suppressed the responses to TRANCE. These results demonstrate that TRANCE has an inflammatory action and may play a role in the pathogenesis of inflammation-related diseases.

Base Sequence↗

PubMed2005

Ischemia-induced changes of platelet endothelial cell adhesion molecule-1 in the hippocampal CA1 region in gerbils.

We observed chronological changes of platelet endothelial cell adhesion molecule-1 (PECAM-1), final mediator of neutrophil transendothelial migration, immunoreactivity, and protein level in the gerbil hippocampus proper after 5 min of transient ischemia. One day after ischemic insult, PECAM-1 immunoreactivity and protein level increased slightly in the hippocampus proper. Thereafter, PECAM-1 immunoreactivity and protein level increased significantly in the hippocampus proper by 4 days after ischemic insult. Especially, PECAM-1 in the hippocampal CA1 region was higher than that in the CA2/3 region. Five days after ischemic insult, PECAM-1 immunoreactivity decreased compared to the 4 days post-ischemic group. However, the RNA levels of PECAM-1 in the hippocampus proper were significantly decreased in the 4 days post-ischemic groups compared to that in the sham-operated group. This result suggests that the increase of PECAM-1 and decrease of PECAM-1 RNA in the CA1 region 4 days after ischemia may be associated with transmigration of neurotrophil.

Animals↗

PubMed2005

Calbindin D-28k is expressed in the microvascular basal lamina in the ventral horn at early time after transient spinal cord ischemia in the rabbit.

Much evidence has been accumulated that the increased expression of calbindin D-28k (CB) is involved in the blockade of calcium-evoked excitotoxicity in cerebral ischemia. We investigated the expression of CB in the basal lamina of microvessels in the ventral horn of the rabbit spinal cord after transient spinal cord ischemia. Spinal cord sections at the level of L7 were immunostained using monoclonal antibody raised against CB at light and electron microscopic levels. CB immunoreactivity was detected in the basal lamina of microvessels at 30 min after ischemic insult. By 3 h after ischemia, CB immunoreactivity was increased in the basal lamina of the microvessels. CB immunoreactivity began to decrease at 6 h after ischemia and nearly disappeared at 48 h after ischemic insult. For calcium detection in the blood vessels of spinal cord, we conducted an alizarin red staining. Alizarin red reactivity was detected in some microvessels at 3 h after ischemic insult. Our results suggest that the ectopic expression of CB in the microvascular basal laminae may be associated with the buffering of calcium in the endothelial cells of microvessels after ischemic damage.

Animals↗

PubMed2005

Effects of GABAergic transmissions on the immunoreactivities of calcium binding proteins in the gerbil hippocampus.

Although reduced calcium binding protein (CBP) immunoreactivities in the epileptic hippocampus have been well established, it has been controversial that these changes may directly indicate neuronal degeneration. In the present study, therefore, we investigated CBP expressions in the gerbil hippocampus following treatment with gamma-aminobutyric acid (GABA) receptor antagonists in order to assess whether altered CBP expressions are the result of either abnormal excitation or indicative of neuronal damage/degeneration. Seizure-sensitive (SS) gerbils showed a loss/decline of CBP immunoreactivities in some hippocampal neurons as compared with seizure-resistant (SR) gerbils. In muscimol (GABA(A) receptor agonist) treated SS gerbils, expression levels of CBP were enhanced as compared with saline-treated SS gerbils. Bicuculline (a GABA(A) receptor antagonist) treatment markedly reduced CBP immunoreactivities in hippocampal neurons of the SR gerbil. Baclofen (a GABA(B) receptor agonist) treatment increased CBP immunoreactivities in the hippocampus of SS gerbils, although its effect was lower than that of muscimol treatment. Moreover, phaclofen (GABA(B) receptor antagonist) treated SR gerbil showed reduction in calbindin D-28K immunoreactivity, not parvalbumin immunoreactivity, in the hippocampus. These findings therefore suggest that reduced CBP immunoreactivities may be the consequence of abnormal discharge caused by loss of GABAergic inhibition rather than an indication of the neuronal damage/degeneration.

Animals↗

PubMed2005

Enhanced transduction of Cu,Zn-superoxide dismutase with HIV-1 Tat protein transduction domains at both termini.

The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.

Animals↗

PubMed2005

Copper chaperone for Cu,Zn-SOD supplement potentiates the Cu,Zn-SOD function of neuroprotective effects against ischemic neuronal damage in the gerbil hippocampus.

In the present study, we investigated the chronological alterations in SOD1 and its copper chaperone (chaperone for superoxide dismutase, CCS) immunoreactivities and their neuroprotective effects against neuronal damage in the gerbil hippocampus after 5 min of transient forebrain ischemia. SOD1 and CCS immunoreactivities were significantly increased in the stratum pyramidale of the CA1 region at 24 and 12 h after ischemic insult, respectively. At 24 h after ischemic insult, the SOD1 and CCS immunoreactivities were colocalized in the CA1 pyramidal cells of the stratum pyramidale. Thereafter, their immunoreactivities were significantly decreased in the CA1 region. To elucidate the effects of CCS or CCS/SOD1, we constructed the expression vectors PEP-1-SOD and PEP-1-CCS. In the CCS-treated group and the CCS/SOD1-treated group, 43.9 and 78.9% pyramidal cells, respectively, compared to the sham-operated group, were stained with cresyl violet 5 or 7 days after ischemic insult. The distribution pattern of active astrocytes and microglia in the PEP-CCS/SOD1-treated group 5 days after ischemic insult was similar to that of the sham-operated group. In addition, the SOD activity in the PEP-CCS- or PEP-CCS/SOD1-treated group was maintained by 10 days after ischemic insult. The SOD activity was higher in the PEP-CCS/SOD1-treated group vs the CCS-treated group. These results suggest that the enhanced expression of SOD1 and CCS may be related to compensatory mechanisms against ischemic damage and that cotreatment with CCS and SOD1 has a greater neuroprotective effect than treatment with CCS or SOD1 in isolation.

Animals↗

PubMed2005

Chronological changes of neurofilament 200 kDa immunoreactivity in the lateral olfactory tract after transient forebrain ischemia in gerbils.

This study was carried out to investigate the transient ischemia-induced changes of neurofilament 200 kDa (NF-200) immunoreactivity and protein content in the gerbil lateral olfactory tract (LOT) after 5 min of transient forebrain ischemia. Weak NF-200 immunoreactivity was detectable in the LOT in the sham-operated group. In this group, a few somata of mitral cells showed weak NF-200 immunoreactivity. One day after transient ischemia, NF-200 immunoreactivity in the LOT was increased significantly. NF-200 immunoreactivity in the LOT by 15 days after ischemia was similar to that in the 1 day post-ischemic group. In this time period, strong NF-200 immunoreactivity was expressed in the mitral cell processes, but the immunoreactivity in the mitral cell somata was significantly decreased. Thereafter, NF-200 immunoreactivity in the LOT was decreased significantly by 30 days after ischemic insult. At this time after ischemia, NF-200 immunoreactivity in the mitral cell dendrites was significantly decreased. The result of Western blot study showed that the pattern of NF-200 expression was similar to that of immunohistochemistry after ischemia-reperfusion. Our result suggests that changes of NF-200 protein in the gerbil LOT may be related to response to ischemic damage and that the axonal transport followed transient ischemia may be disturbed.

Animals↗

PubMed2005

Tyrosine kinase A but not phosphacan/protein tyrosine phosphatase-zeta/beta immunoreactivity and protein level changes in neurons and astrocytes in the gerbil hippocampus proper after transient forebrain ischemia.

In the present study, ischemia-related changes in tyrosine kinase A (trkA) and phosphacan/protein tyrosine phosphatase-zeta/beta (PTP-zeta/beta) immunoreactivities and protein contents were examined in the hippocampus proper after transient forebrain ischemia for 5 min in a gerbil model. Our investigations showed that ischemia-induced changes occurred in trkA immunoreactivity in the hippocampal CA1 region, but not in the CA2/3 region of the hippocampus proper. In the sham-operated group, trkA immunoreactivity was barely detectable. trkA immunoreactivity increased from 30 min after ischemia and peaked at 12 h. Four days after ischemic insult, trkA immunoreactivity was observed in GFAP-immunoreactive astrocytes in the strata oriens and radiatum. In addition, we found that ischemia-related changes in trkA protein content were similar to immunohistochemical changes. On the other hand, PTP-zeta/beta immunoreactivities in the hippocampus proper were unaltered by forebrain ischemia. These results suggest that chronological changes of trkA after transient forebrain ischemia may be associated with an ischemic damage compensatory mechanism in CA1 pyramidal cells.

Adaptation, Physiological↗

PubMed2005

Transduced Tat-SOD fusion protein protects against ischemic brain injury.

Reactive oxygen species (ROS) are implicated in reperfusion injury after transient focal cerebral ischemia. The antioxidant enzyme, Cu,Zn-superoxide dismutase (SOD), is one of the major means by which cells counteract the deleterious effects of ROS after ischemia. Recently, we reported that when Tat-SOD fusion protein is transduced into pancreatic beta cells it protects the beta cells from destruction by relieving oxidative stress in ROS-implicated diabetes (Eum et al., 2004). In the present study, we investigated the protective effects of Tat-SOD fusion protein against neuronal cell death and ischemic insults. When Tat-SOD was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Immunohistochemical analysis revealed that Tat-SOD injected intraperitoneally (i.p.) into mice has access to various tissues including brain neurons. When i.p. injected into gerbils, Tat-SOD prevented neuronal cell death in the hippocampus in response to transient fore-brain ischemia. These results suggest that Tat-SOD provides a strategy for therapeutic delivery in various hu-man diseases, including stroke, related to this anti-oxidant enzyme or to ROS.

Animals↗

PubMed2005

Production and characterization of monoclonal antibodies against human ceruloplasmin.

Ceruloplasmin (CP) is the major plasma antioxidant and copper transport protein. Monoclonal antibodies (mAbs) against human CP were produced and characterized. A total of five hybridoma cell lines were established (CP2, CP10, CP20, CP25, CP30). From the epitope mapping analysis, two subgroups of mAbs recognize different peptide fragments were identified. When the purified CP was incubated with the mAbs, the ferroxidase activity of CP was inhibited up to a maximum 57 %. Immunoblotting with various tissue homogenates indicated that all the mAbs specifically recognize a single protein band of 130 kDa. They also appear to be extensively cross-reactive among different mammalian including human and avian sources. These results demonstrated that only one type of immunologically similar CP is present in all of the mammalian tissues including human. The CP mAbs could be of great benefit to design the diagnostic kit for CP-related diseases such as Wilson's disease.

Animals↗

PubMed2005

Calcium-independent CaMKII activity is involved in ginsenoside Rb1-mediated neuronal recovery after hypoxic damage.

Recent studies have indicated that Ginsenoside Rb1, one of the major components of ginseng root, may play an important role in protecting cells from damage. Here, we investigated the neuroprotective activity of Rb1 after hypoxic injury in young rats. About 50% animals were dead by exposing hypoxic condition three times in three consecutive days. Then, the pretreatment with Rb1 prior to hypoxic stimulation reduced animal death to 12%, and also significantly reduced the recovery time from hypoxia-related, compromised symptoms in survived animals. Rb1 also significantly reduced levels of lactate dehydrogenase (LDH) release from primary hippocampal neurons which were maintained at low oxygen concentration, indicating increased neuronal survival by Rb1. Ca(2+)/calmodulin-dependent kinase II (CaMKII) activity in the hippocampal tissues of hypoxia-induced rats was decreased to about 50% of the control animal. Then Rb1-treatment prior to hypoxic stimulation significantly elevated Ca(2+)-independent kinase II activity when measured 48 hr after hypoxic stimulation. Thus, the present data suggest that calcium independent CaMKII activity may be involved in the process of ginsenoside Rb1-mediated recovery of neuronal cells after hypoxic damage.

Animals↗

PubMed2005

Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells.

Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B6 to pyridoxal-5'-phosphate (PLP). A human brain PK gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-PK fusion protein. The expressed and purified Tat-PK fusion proteins transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced Tat-PK proteins showed catalytic activity and are stable for 48 h. The intracellular concentration of PLP, which is known as a biologically active form of vitamin B6, was increased by pre-treatment of Tat-PK to the PC12 cells. Those results suggest that the transduction of Tat-PK fusion protein can be one of the ways to regulate the PLP level and to replenish this enzyme in the various neurological disorders related to vitamin B6.

Animals↗

PubMed2005

Elevated voltage gated Cl- channel expression enhances fast paired-pulse inhibition in the dentate gyrus of seizure sensitive gerbil.

The present study was performed to determine whether the voltage gated Cl- channel (CLC) expression is altered in the hippocampus of seizure sensitive (SS) gerbils, and to identify the strong fast paired-pulse inhibition in the dentate gyrus of SS gerbils is associated with altered CLC expression. In the hippocampal proper and the granule cell layer of the dentate gyrus of the SS gerbils, strong CLC-2 immunoreactivity was detected, as compared with seizure resistant (SR) gerbils. In addition, CLC-3 immunoreactivity was observed in the CA1-3 pyramidal cells, and the granule cell and the molecular layer of the dentate gyrus in the SS gerbils, whereas its immunoreactivity was rarely detected in the SR gerbils. However, CLC-3 immunoreactivity in the mossy fiber was reduced, as compared with SR gerbils. Moreover, infusion of the potential CLC inhibitor (4,4'-diisothiocyanostibene-2,2'-disulfanic acid, DIDS) reduced fast paired-pulse inhibition in the dentate gyrus of SS gerbils, although evoked responses in the dentate gyrus between SR and SS gerbils were similarly detected. These findings suggest that enhancement of CLC expression in the dentate gyrus of SS gerbils may be one of the compensatory responses for reduced GABA(A) receptor-mediated fast postsynaptic inhibitory potentials.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗