Biomedical subjects

Moo Ho Won

Publications and source records attributed to Moo Ho Won.

At least 73 records · Page 4Linked to original sources

PubMed2005

Seizure-induced changes of mineralocorticoid and glucocorticoid receptors in the hippocampus in seizure sensitive gerbils.

Abnormal corticosteroid hormone levels during stress and resultant mineralocorticoid receptor (MR)/glucocorticoid receptor (GR) imbalance enhance the vulnerability of specific hippocampal neurons. In the present study, we investigated the distribution of MR and GR in seizure resistant (SR) and seizure sensitive (SS) gerbils, and observed the seizure-induced changes of MR and GR in the hippocampus of SS gerbils using immunohistochemistry and western blot analysis. MR and GR immunoreactivities were higher in the SS pre-seizure gerbils than that in SR gerbils. In the SR gerbils, the immunodensity of GR was high compared to that of MR. The changes of MR and GR immunoreactivities were significant in the stratum pyramidale of the hippocampal CA1 region and the infrablade of the dentate gyrus after seizure on-set. MR immunoreactivity in the CA1 region was significantly increased at 12h after seizure on-set, thereafter MR immunoreactivity was decreased. MR immunoreactivity in the dentate gyrus was decreased time-dependently after seizure on-set. GR immunoreactivity was decreased in the CA1 region and dentate gyrus time-dependently after seizure on-set. At 12h after seizure on-set, differences in MR and GR immunodensity diminished in the CA1 region and dentate gyrus. This imbalance of MR and GR immunoreactivity in these regions may be associated with seizure generation in the Mongolian gerbil, which is a hereditary seizure model.

Animals↗

PubMed2005

Ischemia-related changes in galanin expression in the dentate hilar region after transient forebrain ischemia in gerbils.

Although galanin (GAL) protects hippocampal neurons from ischemic damage, no study has examined ischemia-related changes in endogenous GAL in the hippocampal dentate gyrus. We investigated the chronological changes of GAL, well-known as the potassium channel opener, expression in the dentate gyrus at various times after 5 min of transient forebrain ischemia in gerbils. A few GAL-immunoreactive (IR) neurons were found in the polymorphic layer of the sham-operated group. Three hours after ischemia-reperfusion, the pattern of GAL immunoreactivity was similar to that of the sham-operated group and the number of GAL-IR neurons and immunoreactivity were highest 12 h after ischemic insult. At this time, GAL-IR neurons in the polymorphic layer showed strong GAL immunoreactivity. Thereafter, GAL-IR neurons and immunoreactivity significantly decreased in the dentate hilar region. Four days after ischemic insult, GAL-IR neurons were not detectable. In addition, the results of a Western blot study showed a pattern of GAL expression similar to the immunohistochemical changes. GAL protein content also was highest 12 h after ischemia. In conclusion, the increased expression of endogenous GAL in the dentate gyrus after ischemia is related to response to the ischemic damage.

Animals↗

PubMed2005

Spontaneous ophthalmic diseases in 586 New Zealand white rabbits.

The purpose of this study was to investigate spontaneous eye disease in New Zealand White (NZW) rabbits, which are commonly used for toxicity tests, and to provide reference materials for pharmaceutical companies and research centers. A total of 586 NZW rabbits were randomly chosen without sex preference and were examined using ocular equipment, including a direct ophthalmoscope, an indirect ophthalmoscope, a slit-lamp biomicroscope, a focal illuminator, and a fundus camera. This study showed that the incidence rate of temporary cataracts, regarded as a change within normal variation, was 0.5% in the NZW rabbits. Regarding abnormal ophthalmic disease, blepharitis was the most commonly observed ocular disease. Other findings included cataract, conjunctivitis, choroidal hypoplasia, keratitis, corneal scarring, eyelid laceration, posterior synechiae, uveitis, dacryocystitis, and persistent pupillary membrane. In total, the incidence rate of ophthalmic diseases was 9.6%. Based on sex and age distributions, females had more ocular diseases than males, and rabbits were less susceptible to eye diseases as they got older. In this study, photographs were taken to document findings, such as normal fundus, normal variations, ophthalmic disease, and histopathologic examination.

Animals↗

PubMed2004

Site-directed mutagenesis of human brain GABA transaminase: lysine-357 is involved in cofactor binding at the active site.

gamma-Aminobutyrate transaminase (GABA-T), a key enzyme of the GABA shunt, converts the major inhibitory neurotransmitter, GABA, to succinic semialdehyde. Although GABA-T is a pivotal factor implicated in the pathogenesis of various neurological disorders, its function remains to be elucidated. In an effort to clarify the structural and functional roles of specific lysyl residue in human brain GABA-T, we constructed human brain GABA-T mutants, in which the lysyl residue at position 357 was mutated to various amino acids including asparagine (K357N). The purified mutant GABA-T enzymes displayed neither catalytic activity nor absorption bands at 330 and 415 nm that are characteristic of pyridoxal-5'-phosphate (PLP) covalently linked to the protein. The wild type apoenzyme reconstituted with exogenous PLP had catalytic activity, while the mutant apoenzymes did not. These results indicate that lysine 357 is essential for catalytic function, and is involved in binding PLP at the active site.

4-Aminobutyrate Transaminase↗

PubMed2004

Elevated voltage-gated Ca2+ channel immunoreactivities in the hippocampus of seizure-prone gerbil.

In present study, we investigated voltage-gated Ca2+ channel (VGCC) expressions in the hippocampus of the Mongolian gerbil and its association with different sequelae of spontaneous seizures, in an effort to identify the epileptogenesis in this animal. In the hippocampus of pre-seizure seizure sensitive (SS) gerbils, VGCC subunit expressions were significantly elevated, as compared with seizure-resistant (SR) gerbils. In 3 h postictal group, the alteration of VGCC expressions showed regional- and neuronal-specific manners; VGCC immunoreactivities in principal neurons were markedly decreased; however, their immunoreactivities in interneurons were significantly elevated. These results are the first comprehensive description of the distribution of VGCC immunoreactivities in the normal and epileptic hippocampus of gerbils, and suggest that these alterations in the hippocampus of the SS gerbil may be related with tissue excitability and have a role in modulating recurrent excitation following seizures.

Animals↗

PubMed2004

Protein disulfide isomerase immunoreactivity and protein level changes in neurons and astrocytes in the gerbil hippocampal CA1 region following transient ischemia.

We investigated the temporal and spatial alterations of protein disulfide isomerase (PDI) immunoreactivity and protein level in the hippocampus proper after 5 min transient forebrain ischemia in gerbils. PDI immunoreactivity was significantly altered in the hippocampal CA1 region. PDI immunoreactivity in the sham-operated animals was found in non-pyramidal cells. At 30 min after ischemia, PDI immunoreactivity was shown in the pyramidal cells of the stratum pyramidale (SP): the PDI immunoreactivity in the pyramidal cells was increased up to 12 h after ischemia. Thereafter PDI immunoreactivity was decreased, and the PDI immunoreactivity was shown in non-pyramidal cells 2 days after ischemia. Four to 5 days after ischemia, almost pyramidal cells in the CA1 region were lost because the delayed neuronal death occurred. At this time period, PDI immunoreactivity was expressed in some astrocytes as well as some neurons. The results of the Western blot analysis were consistent with the immunohistochemical data. These findings suggest that increase of PDI in pyramidal cells may play a critical role in resistance to ischemic damage at early time after ischemic insult, and that expression of this protein in astrocytes at late time after ischemic insult is partly implicated in the acquisition of tolerance against ischemic stress.

Adaptation, Physiological↗

PubMed2004

Elevated substance P (NK-1) receptor immunoreactivity in the cerebellum of seizure prone gerbil.

In the present study, we performed a comparative analysis of the distribution of substance P (SP) receptor (NK-1) immunoreactivity in order to determine the characteristics of the SP system in the cerebelli of rat and gerbils. In the rat cerebellar cortex, only a few Purkinje cells exhibited weak NK-1 receptor immunoreactivity. Similar to the case of rat, NK-1 receptor immunoreactivity in the cerebellar cortex of seizure resistant (SR) gerbils was rarely detected. In contrast, in the cerebellar cortex of seizure sensitive (SS) gerbils, dendrites and cell bodies of Purkinje cell showed strong NK-1 receptor immunoreactivity. Similar to the cerebellar cortex, little NK-1 receptor immunoreactivity in deep cerebellar nuclei was observed in the rat. In SR gerbils, however, deep cerebellar nuclei showed weak NK-1 receptor immunoreactivity. NK-1 receptor immunoreactivity in the deep cerebellar nuclei of SS gerbils was markedly increased, as compared with SR gerbils. Based on the present data, we suggest that the SP system of cerebellar circuit in gerbil are different from rat, and over-expression of NK-1 receptor immunoreactivity in Purkinje cells of SS gerbils may be relevant to Purkinje cell loss induced by seizure activity.

Animals↗

PubMed2004

In vivo protein transduction: biologically active intact pep-1-superoxide dismutase fusion protein efficiently protects against ischemic insult.

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 denatured Tat-SOD fusion protein is transduced into cells and skin tissue. Moreover, PEP-1 peptide, which has 21 amino acid residues, is a known carrier peptide that delivers full-length native proteins in vitro and in vivo. In the present study, we investigated the protective effects of PEP-1-SOD fusion protein after ischemic insult. A human SOD gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-SOD fusion protein. The expressed and purified fusion proteins were efficiently transduced both in vitro and in vivo with a native protein structure. Immunohistochemical analysis revealed that PEP-1-SOD injected intraperitoneally (i.p.) into mice can have access into brain neurons. When i.p.-injected into gerbils, PEP-1-SOD fusion proteins prevented neuronal cell death in the hippocampus caused by transient forebrain ischemia. These results suggest that the biologically active intact forms of PEP-1-SOD provide a more efficient strategy for therapeutic delivery in various human diseases related to this antioxidant enzyme or to ROS, including stroke.

Animals↗

PubMed2004

Cysteine-321 of human brain GABA transaminase is involved in intersubunit cross-linking.

Gamma-aminobutyrate transaminase (GABA-T), a key homodimeric enzyme of the GABA shunt, converts the major inhibitory neurotransmitter GABA to succinic semialdehyde. We previously overexpressed, purified and characterized human brain GABA-T. To identify the structural and functional roles of the cysteinyl residue at position 321, we constructed various GABA-T mutants by site-directed mutagenesis. The purified wild type GABA-T enzyme was enzymatically active, whereas the mutant enzymes were inactive. Reaction of 1.5 sulfhydryl groups per wild type dimer with 5,5 cent-dithiobis-2-nitrobenzoic acid (DTNB) produced about 95% loss of activity. No reactive -SH groups were detected in the mutant enzymes. Wild type GABA-T, but not the mutants, existed as an oligomeric species of Mr = 100,000 that was dissociable by 2-mercaptoethanol. These results suggest that the Cys321 residue is essential for the catalytic function of GABA-T, and that it is involved in the formation of a disulfide link between two monomers of human brain GABA-T.

4-Aminobutyrate Transaminase↗

PubMed2004

Expression and changes of galanin in neurons and microglia in the hippocampus after transient forebrain ischemia in gerbils.

In the present study, we investigated chronological changes of galanin (GAL), well known as the potassium channel opener, immunoreactivity and GAL protein level in the hippocampus of the gerbil at the various times after 5 min transient forebrain ischemia. In the sham-operated group, weak GAL immunoreactivity was found in non-pyramidal cells. At 12 h after ischemia-reperfusion, the number of GAL-immunoreactive neurons and GAL immunoreactivity were significantly increased in the hippocampus compared to 3 h after ischemic insult, especially in the hippocampal CA1 region. Thereafter the number of GAL-immunoreactive neurons and GAL immunoreactivity decrease time-dependently in the hippocampus. Four days after transient ischemia, GAL immunoreactivity was low as compared with the sham-operated group. At this time point after ischemic insult, GAL immunoreactivity was shown in microglia in the CA1 region because delayed neuronal death happened in the CA1 pyramidal cells. The result of Western blot showed the pattern of GAL expression similar to that of immunohistochemical data. These results suggest that the early increase of GAL in the CA1 pyramidal cells may be associated with the reduction of the excitotoxic damage, that long-lasting enhanced expression of endogenous GAL at 12 h-2 days after ischemia may be associated with efflux of potassium ion into the extracellular space, and that GAL expression in microglia 4 days after ischemia may be associated with reduction of ischemic damage.

Analysis of Variance↗

PubMed2004

Expression and changes of calbindin D-28k immunoreactivity in the ventral horn after transient spinal cord ischemia in rabbits.

We examined ischemia-related changes of calbindin D-28k (CB) immunoreactivity in L(7) of the spinal ventral horn after transient spinal cord ischemia in rabbits. In the sham-operated group, CB immunoreactivity was not present in the spinal ventral horn, but CB immunoreactivity was detectable in the dorsal horn. CB immunoreactivity was detectable in the ventral horn at 30 min after ischemia: the CB immunoreactivity was found in glial cells identified as astrocytes. At 1 h after ischemia, CB immunoreactivity was highest and present at a few somata located in the lamina VII as well as many glial cells. CB immunoreactivity was lower in the lamina VII at 3 h after ischemia compared to 1 h post-ischemic group. By 2 days after ischemia, CB immunoreactivity was decreased in this region. In addition, the result of Western blot result showed the pattern of CB expression similar to that of immunohistochemistry. In conclusion, the ischemia-related changes of CB immunoreactivity in neurons and glial cells in the ischemic spinal ventral horn in rabbits may be related to modulation of intracellular calcium following transient ischemia.

Animals↗

PubMed2004

Neuroprotective effects of grape seed extract on neuronal injury by inhibiting DNA damage in the gerbil hippocampus after transient forebrain ischemia.

Grape seed extract (GSE) possess cardioprotective abilities by functioning as in vivo antioxidants and by virtue of their ability to directly scavenge ROS including hydroxyl and peroxyl radicals. In the present study, we investigated the neuroprotective effects of grape seed extract (GSE) in the gerbil hippocampus after 5 min transient forebrain ischemia. Neuronal cell density in GSE-treated ischemic animals was significantly increased as compared with vehicle-treated ischemic animals 4 days after ischemic insult. In the GSE-treated groups, about 60% of pyramidal cells of the sham-operated group were stained with cresyl violet 4 days after ischemic insult. In this study, we found that GSE had neuroprotective effects on neuronal injury by inhibiting DNA damage in the CA1 region after ischemia. In vehicle-treated groups, 8-hydroxy-2'-deoxyguanosine (8-OHdG) immunoreactivity was significantly changed time-dependently, whereas the immunoreactivity in the GSE-treated group was similar to the sham-operated group. In addition, we confirmed that astrocytes and microglia did not show significant activation in the CA1 region 4 days after ischemia-reperfusion, because many CA1 pyramidal cells were not damaged. Therefore, these results suggest that GSE can protect ischemic neuronal damage by inhibiting DNA damage after transient forebrain ischemia.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed2004

Very delayed neuronal loss occurs in the glomerular layer of the main olfactory bulb following transient ischemia in gerbils.

Olfactory dysfunction could happen following various insults such as ischemic-hypoxic state. Neurons of the main olfactory bulb (MOB) are resistant to ischemic damage. In the present study, we investigated the ischemia-related changes of neurons and glial cells in the glomerular layer (GL) of the gerbil MOB after transient ischemia. The number of NeuN-immunoreactive neurons became to decrease from 10 days after ischemic insult. Fifteen days after ischemic insult, astrocytes and microglia were increased in number. By 60 days after ischemia, NeuN-immunoreactive neurons were significantly decreased by 42% per glomerulus. At this time period, astrocytes and microglia were pronouncedly increased. This result indicates that neuronal loss must be much delayed in the GL following transient ischemia.

Animals↗

PubMed2004

Age-related changes of gamma-aminobutyric acid transaminase immunoreactivity in the hippocampus and dentate gyrus of the Mongolian gerbil.

We investigated the age-related changes of gamma-aminobutyric acid transaminase (GABA-T, a GABA degradation enzyme) in the hippocampus and dentate gyrus of the gerbil at postnatal month 1 (PM 1), PM 3, PM 6, PM 12, and PM 24. Age-related changes of GABA-T immunoreactivity were distinct in the hippocampal CA1 region and in the dentate gyrus. GABA-T immunoreactivity was weak at PM 1, but at PM 3, it had increased significantly, and then increased further. Between PM 6 and PM 12, strong GABA-T immunoreactivity was found in nonpyramidal cells (GABAergic) in the stratum pyramidale of the CA1 region, and at PM 6, strong GABA-T immunoreactivity was found in neurons of the dentate gyrus subgranular zone. At PM 24, CA1 pyramidal cells showed strong GABA-T immunoreactivity. Western blot analysis showed a pattern of GABA-T expression similar to that shown by immunohistochemistry at various ages. In conclusion, our results suggest that the age-related changes of GABA-T provide important information about the aged brain with GABA dysfunction.

4-Aminobutyrate Transaminase↗

PubMed2004

Chronological alterations of neurofilament 150 immunoreactivity in the gerbil hippocampus and dentate gyrus after transient forebrain ischemia.

In this study, we observed the chronological alterations of neurofilament 150 (NF-150) immunoreactivity in the gerbil hippocampus and dentate gyrus after 5 min transient forebrain ischemia. NF-150 immunoreactivity in the sham-operated group was mainly detected in mossy fibers and in the hilar region of the dentate gyrus. NF-150 immunoreactivity and protein contents of NF-150 and RT 97 (polyphosphorylation epitopes of neurofilament) were significantly decreased at 15 min after ischemic insult. Between 30 min and 12 h after ischemic insult, NF-150 immunoreactivity and protein content were significantly increased as compared with the sham-operated group. Thereafter, NF-150 immunoreactivity and protein content started to decrease. At 12 h after ischemic insult, unlike dentate gyrus, NF-150 immunoreactivity increased in pyramidal cells of the CA1 region. Thereafter, NF-150 immunoreactivity in the CA1 region started to decrease, and 4 days after ischemic insult, NF-150 immunoreactivity nearly was similar to that of the sham-operated group. These biphasic patterns of NF-150 immunoreactivity in the hippocampus and dentate gyrus are reverse correlated with that of the intracellular calcium influx. For calcium detection in the CA1 region, we also conducted alizarin red staining. Alizarin red positive neurons were detected in some neurons at 15-30 min after ischemic insult. At 12 h after ischemia, alizarin red positive neurons were decreased. Thereafter, alizarin red positive neurons started to decrease, but alizarin positive neurons were significantly increased in dying neurons 4 days after ischemia. These results suggest that ischemia-related changes of NF-150 expression may be caused by the calcium following transient forebrain ischemia.

Animals↗

PubMed2004

Changes in parvalbumin immunoreactivity in the parietofrontal cortex after transient forebrain ischemia in the Mongolian gerbil.

We investigated the changes in parvalbumin (PV)-immunoreactive (IR) neurons in the parietofrontal cortex after transient forebrain ischemia. In the sham-operated group, PV-IR neurons were present in all layers of the parietofrontal cortex except layer I. Shortly after ischemia the number of PV-IR neurons in layer II/III first increased, and then declined dramatically 12 h after ischemic insult, followed by a second increase after 2 days. At this time the PV immunoreactivity was very weak and only present in the peripheral neuronal cytoplasm. The reversible increase in the number of PV-IR neurons and in the level of their immunoreactivity could result from a transient ischemia-induced increase in intracellular calcium. This pattern of expression was particularly pronounced in layer II/III of the parietofrontal cortex, suggesting that these neurons are especially\susceptible to ischemic insult.

Animals↗

PubMed2004

GABAA, not GABAB, receptor shows subunit- and spatial-specific alterations in the hippocampus of seizure prone gerbils.

In the present study, we investigated site-specific expressions of GABA(A) and GABA(B) receptor subunits in the seizure-sensitive (SS) and seizure-resistant (SR) gerbil hippocampus to elucidate the function of the gamma-aminobutyric acid (GABA) receptor in seizure activity in this animal. There were no differences of the immunoreactivities of GABA(B) receptor and some GABA(A) receptor subunits (alpha3, alpha4, pan beta and delta) in the hippocampus between SR and SS gerbils. The alpha1 subunit expression was mainly detected in interneurons of stratum radiatum and hilar region of dentate gyrus in the SR gerbil. However, in SS gerbil, interneurons were nearly devoid of alpha1 subunit immunoreactivity and mainly detected in the molecular layer of dentate gyrus. In SR gerbil, alpha2 subunit immunoreactivity was detected in Ammon's horn, particularly in the CA2 region. In SS gerbil, granule cell layer of the dentate gyrus in SS gerbil showed strong alpha2 subunit immunoreactivity. The distribution of alpha5 and gamma2 subunit immunoreactivity in the hippocampus was similarly detected in SR and SS gerbil. However, alpha5 immunodensity of SR gerbil was slightly lower than that of SS gerbil in CA1 region and was slightly strong than that of SS gerbil in subiculum. These differences in distribution of GABA(A) receptor, not GABA(B) receptor, in the SR and SS gerbil hippocampus may indicate that abnormal hyperactive neuronal discharges are occurred in SS gerbil, which presumably result in spontaneous and repetitive seizure activity in this animal.

Animals↗

PubMed2004

GABA(B) receptor-mediated regulation of P2X7 receptor expression in the gerbil hippocampus.

In the present study, the P(2)X(7) receptor expression in the gerbil hippocampus and GABA-mediated responses of its expression was investigated in order to identify the roles of the P(2)X(7) receptor on seizure activity and recovery mechanisms. P(2)X(7) receptor immunoreactivity in seizure-resistant (SR) gerbils was similar to that in pre-seizure group of seizure-sensitive (SS) gerbils. The administration of baclofen, a GABA(B) receptor agonist, P(2)X(7) receptor immunoreactivity was decreased in the mossy fiber, compared with that of non-treated gerbils, whereas treatment with phaclofen, a GABA(B) receptor antagonist, elevated P(2)X(7) receptor expression. Neither the treatments with GABA(A) receptor agonist nor antagonist affected P(2)X(7) receptor expression in the hippocampus. These findings suggest that altered P(2)X(7) receptor expression may not be involved in the epileptogenesis or seizure activity in gerbils, and presynaptic GABA(B) receptor-mediated actions may be closely related with the regulation of P(2)X(7) receptor expression in the gerbil hippocampus.

Analysis of Variance↗