Bac k g r o u n d: Polybrominated diphenyl ethers (PBDEs) have been used as flame retardants and are becoming a ubiquitous environmental contaminant. Adverse effects in the developing brain are of great health concern. Obj e c t i v e: We... more
Bac k g r o u n d: Polybrominated diphenyl ethers (PBDEs) have been used as flame retardants and are becoming a ubiquitous environmental contaminant. Adverse effects in the developing brain are of great health concern. Obj e c t i v e: We investigated the effect of PBDEs/hydroxylated PBDEs (OH‑PBDEs) on thyroid hormone (TH) receptor (TR)-mediated transcription and on TH-induced dendrite arborization of cerebellar Purkinje cells. Met h o d s: We examined the effect of PBDEs/OH‑PBDEs on TR action using a transient transfection-based reporter gene assay. TR–cofactor binding was studied by the mammalian two-hybrid assay, and TR–DNA [TH response element (TRE)] binding was examined by the liquid chemiluminescent DNA pull-down assay. Chimeric receptors generated from TR and glucocorticoid receptor (GR) were used to identify the functional domain of TR responsible for PBDE action. The change in dendrite arborization of the Purkinje cell in primary culture of newborn rat cerebellum was also ...
Pratt, T., Tian, N.M., Simpson, T.I., Mason, J.O., Price, D.J., 2004. The winged helix transcription factor Foxg1 facilitates retinal ganglion cell axon crossing of the ventral midline in the mouse. Development 131, 3773–3784. Xuan, S.,... more
Pratt, T., Tian, N.M., Simpson, T.I., Mason, J.O., Price, D.J., 2004. The winged helix transcription factor Foxg1 facilitates retinal ganglion cell axon crossing of the ventral midline in the mouse. Development 131, 3773–3784. Xuan, S., Baptista, C.A., Balas, G., Tao, W., Soares, V.C., Lai, E., 1995. Winged helix transcription factor BF-1 is essential for the development of the cerebral hemispheres. Neuron 14, 1141–1152.
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From the information of nucleotide sequences and deduced amino acid sequences of flaviviruses including JEV, we can postulate processing mechanisms of a polyprotein translated from single long open reading frame of the genome and... more
From the information of nucleotide sequences and deduced amino acid sequences of flaviviruses including JEV, we can postulate processing mechanisms of a polyprotein translated from single long open reading frame of the genome and mechanisms of construction of antigenic structures of structural proteins with biologically active forms after these proteins are translated. The results of comparative analysis of amino acid sequences among flaviviruses and epitope analysis on the E proteins which are the most important antigens for protective immunity suggest that the E protein of flaviviruses may have a similar structure closely related to each other. PrM and E proteins which had predictable signal sequences upstream on the N terminals were expressed with antigenically active form and molecular size the same as the authentic ones by the recombinant viruses. However, the recombinant viruses which had no such signal sequence expressed unprocessed proteins with antigenically denatured forms...
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Research Interests: Cognitive Science, Biology, Cell Biology, Extracellular Matrix, Axon, and 15 moreCerebellum, Cerebral Cortex, Animals, Central Nervous System, Astrocytes, Analysis of Variance, Lesion, Cell Proliferation, Axons, Antigens, Gene Expression Regulation, fibroblasts, Glial scar, Cell count, and coculture techniques
Research Interests: Microbiology, Medical Microbiology, Biology, Virology, Medicine, and 15 moreCerebrospinal Fluid, Humans, Virus, Sheep, Animals, Monoclonal Antibodies, Japanese encephalitis, Antibody, Microbial Sensitivity Tests, Monoclonal Antibody, Hemagglutination, Japanese Encephalitis Virus, Enzyme Linked Immunosorbent Assay, Antigen, and immunoglobulin M
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Research Interests: Kinetics, Biology, Virology, Membrane Proteins, Medicine, and 15 moreBiological Sciences, Cercopithecus aethiops, Cell line, Virus, Animals, Monoclonal Antibodies, Japanese encephalitis, Fluorescent Antibody Technique, Rats, Monoclonal Antibody, Japanese Encephalitis Virus, Vero cells, Indirect fluorescent antibody, Cell Membrane, and Medical and Health Sciences
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A cDNA clone representing the genome of structural proteins of Japanese encephalitis virus (JEV) was inserted into the thymidine kinase gene of vaccinia virus strains LC16mO and WR under the control of a strong early-late promoter for the... more
A cDNA clone representing the genome of structural proteins of Japanese encephalitis virus (JEV) was inserted into the thymidine kinase gene of vaccinia virus strains LC16mO and WR under the control of a strong early-late promoter for the vaccinia virus 7.5-kilodalton polypeptide. Indirect immunofluorescence and fluorescence-activated flow cytometric analysis revealed that the recombinant vaccinia viruses expressed JEV E protein on the membrane surface, as well as in the cytoplasm, of recombinant-infected cells. In addition, the E protein expressed from the JEV recombinants reacted to nine different characteristic monoclonal antibodies, some of which have hemagglutination-inhibiting and JEV-neutralizing activities. Radioimmunoprecipitation analysis demonstrated that two major proteins expressed in recombinant-infected cells were processed and glycosylated as the authentic PreM and E glycoproteins of JEV. Inoculation of rabbits with the infectious recombinant vaccinia virus resulted ...
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In a study on Fischer rats, all animals infected with Japanese encephalitis virus (JEV) before the age of 13 days died, but animals infected after the age of 14 days did not die, confirming the age-dependent resistance to JEV infection in... more
In a study on Fischer rats, all animals infected with Japanese encephalitis virus (JEV) before the age of 13 days died, but animals infected after the age of 14 days did not die, confirming the age-dependent resistance to JEV infection in the rat brain. A study of the kinetics of JEV infection in the developing rat brain disclosed that JEV antigen disappeared in a particular pattern, i.e., from the deeper layers to the upper layers of the motor cortex, which paralleled neuronal maturation in the cortex. Fifteen-day-old rats, which were resistant to JEV infection, received intracerebral transplants of neurons taken from 19-day embryos. When these animals were infected with JEV after transplantation, viral antigen was detected only in the embryonal neurons soon after transplantation. Thus, it can be concluded that the susceptibility to JEV infection in the rat brain is closely associated with neuronal immaturity.
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Research Interests: Biology, Immunohistochemistry, Medicine, Cell Division, Dengue Virus, and 15 moreBiological Sciences, Cell line, Humans, Mice, Dengue fever, Female, Animals, Animal Model, Dengue, Cell Transplantation, Blood Urea Nitrogen, Mouse Model, Severe combined immunodeficiency, Liver neoplasms, and Medical and Health Sciences
Research Interests: Electron Microscopy, Biology, Public Health, Medicine, Adsorption, and 15 moreCercopithecus aethiops, Cell fusion, Endocytosis, Animals, Glycoproteins, Japanese encephalitis, Activity, Antibody, Monoclonal Antibody, Neutralisation, Japanese Encephalitis Virus, Neutralization, Antibody dependent enhancement, Neutralization tests, and Medical and Health Sciences
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... 21 Yahi, N., Sabatier, JM, Nickel, E, Mabrouk, K., Gonzalez-Scarano, E and Fantini, J., J. Biol. Chem., 269 (1994) 24349. ... Neurol., 23 (1988) 339. 48 Merrill, JE, Koyanagi, Y., Zack, J., Thomas, L., Martin, F. and Chen, ISY, J.... more
... 21 Yahi, N., Sabatier, JM, Nickel, E, Mabrouk, K., Gonzalez-Scarano, E and Fantini, J., J. Biol. Chem., 269 (1994) 24349. ... Neurol., 23 (1988) 339. 48 Merrill, JE, Koyanagi, Y., Zack, J., Thomas, L., Martin, F. and Chen, ISY, J. Virol., 66 (1992) 2217. ...
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We assessed the validity of immunoblotting, immunohistochemistry (IHC), and immunocytochemistry (ICC) to detect anti-neuronal antibodies in an attempt to establish a diagnostic approach for pediatric autoimmune encephalitis. Both IHC and... more
We assessed the validity of immunoblotting, immunohistochemistry (IHC), and immunocytochemistry (ICC) to detect anti-neuronal antibodies in an attempt to establish a diagnostic approach for pediatric autoimmune encephalitis. Both IHC and ICC had higher sensitivity than immunoblotting and could differentiate between antibodies directed towards intracellular and cell surface antigens. There was a significant correlation between the IHC and ICC results. When patients were divided into encephalitis and non-encephalitis groups, there was no difference in the positivity rate and staining pattern of IHC and ICC between them. In conclusion, IHC and ICC are useful methods to screen for anti-neuronal antibodies. A combination of IHC, ICC, and specific cell-based assays is expected to be an efficient approach for the diagnosis of autoantibody-mediated encephalitis.
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Research Interests: Calcium, Multidisciplinary, Insecticides, Brain, Humans, and 11 moreCerebellum, Nicotine, Animals, Neurons, PLoS one, Neurotoxins, Rats, Pyridines, Mecamylamine, Imidazoles, and acetamiprid
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to DPAA at 10 M induced significant increase of mRNA expression of heat shock protein 70 (hsp70), heme oxygenase-1 (hmox1), and superoxide dismutase-1, which suggested that DPAA induced oxidative stress in cultured cerebellar cells.... more
to DPAA at 10 M induced significant increase of mRNA expression of heat shock protein 70 (hsp70), heme oxygenase-1 (hmox1), and superoxide dismutase-1, which suggested that DPAA induced oxidative stress in cultured cerebellar cells. Immunocytochemical analyses revealed that glial fibrillary acidic protein (GFAP) positive cerebellar astrocytes, but not neurons, increased hsp70 and hmox1 protein contents in response to DPAA. Comprehensive analysis of gene expression by cDNA microarray and further validations by real-time PCR revealed siginificant alterations in mRNA expression of some neuroactive and vasoactive peptides by DPAA at 10 M for 24 h; up-regulation of neuropeptide Y (npy), adrenomedullin (adm), chemokine (C-C motif) ligand 2 (ccl2), and fibroblast growth factor 2 (fgf2), and down-regulation of cocaine and amphetamine regulated transcript (cartpt) and somatostatin (sst). GFAP-positive cultured cerebellar astrocytes produced npy, ccl2, and fgf2 peptides in response to DPAA. When adult male Wistar rats were exposed to DPAA for 21 days via drinking water at 100 mg/L, in their cerebellum, mRNA expression of these peptides except for cartpt were significantly altered by DPAA exposure as in DPAAexposed cultured cerebellar cells. These results implicated substantial involvement of these neuroactive and vasoactive peptides produced by astrocytes in the etiology of the cerebellar symptom and abnormal brain blood flow in DPAA-exposed individuals.
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A sensitive, quantitative, short-time, and reproducible focus assay for Japanese encephalitis (JE) virus is described. After 2 or 3 days of incubation, the infected cells were treated with anti-JE virus serum and complement, and... more
A sensitive, quantitative, short-time, and reproducible focus assay for Japanese encephalitis (JE) virus is described. After 2 or 3 days of incubation, the infected cells were treated with anti-JE virus serum and complement, and subsequently stained with trypan blue; then clear foci were produced. This method made it easy to titrate the infectivities not only of all seven JE virus strains tested but also of West Nile (WN), Murray Valley encephalitis (MVE), and St. Louis encephalitis (SLE) viruses using hyperimmune anti-JE virus serum for the latter. Moreover, even cell lines which hardly formed plaques by the agar overlay method easily produced foci within 2 or 3 days by this method.
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Transforming growth factor-β (TGF-β), a multifunctional cytokine, plays a crucial role in wound healing in the damaged central nervous system. To examine effects of the TGF-β signaling inhibition on formation of scar tissue and axonal... more
Transforming growth factor-β (TGF-β), a multifunctional cytokine, plays a crucial role in wound healing in the damaged central nervous system. To examine effects of the TGF-β signaling inhibition on formation of scar tissue and axonal regeneration, the small molecule inhibitor of type I TGF-β receptor kinase LY-364947 was continuously infused in the lesion site of mouse brain after a unilateral transection of the nigrostriatal dopaminergic pathway. At 2 weeks after injury, the fibrotic scar comprising extracellular matrix molecules including fibronectin, type IV collagen, and chondroitin sulfate proteoglycans was formed in the lesion center, and reactive astrocytes were increased around the fibrotic scar. In the brain injured and infused with LY-364947, fibrotic scar formation was suppressed and decreased numbers of reactive astrocytes occupied the lesion site. Although leukocytes and serum IgG were observed within the fibrotic scar in the injured brain, they were almost absent in the injured and LY-364947-treated brain. At 2 weeks after injury, tyrosine hydroxylase (TH)-immunoreactive fibers barely extended beyond the fibrotic scar in the injured brain, but numerous TH-immunoreactive fibers regenerated over the lesion site in the LY-364947-treated brain. These results indicate that inhibition of TGF-β signaling suppresses formation of the fibrotic scar and creates a permissive environment for axonal regeneration.
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Research Interests: Biology, Virology, Medicine, Dengue Virus, Biological Sciences, and 14 moreVirus, Monoclonal Antibodies, Japanese encephalitis, West nile virus, Antibody, Flavivirus, Monoclonal Antibody, Species Specificity, Epitopes, Japanese Encephalitis Virus, Enzyme Linked Immunosorbent Assay, West Nile, Neutralization tests, and Medical and Health Sciences
To identify structures that determine the 90 degree orientation of thin espalier dendritic trees of Purkinje cells with respect to parallel fibers (axonal neurite bundles of granule cells) in the cerebellar cortex, we designed five types... more
To identify structures that determine the 90 degree orientation of thin espalier dendritic trees of Purkinje cells with respect to parallel fibers (axonal neurite bundles of granule cells) in the cerebellar cortex, we designed five types of two-dimensional and three-dimensional cell and tissue cultures of cerebella from postnatal mice and analyzed the orientation of Purkinje cell dendrites with respect to neurite bundles and astrocyte fibers by immunofluorescence double or triple staining. We cultured dissociated cerebellar cells on micropatterned substrates and preformed neurite bundles of a microexplant culture two-dimensionally and in matrix gels three-dimensionally. Dendrites, but not axons, of Purkinje cells extended toward the neurites of granule cells and oriented at right angles two-dimensionally to aligned neurite bundles in the three cultures. In a more organized explant proper of the microexplant culture, Purkinje cell dendrites extended toward thin aligned neurite bundles not only consistently at right angles but also two-dimensionally. However, in the "organotypic microexplant culture," in which three-dimensionally aligned thick neurite bundles mimicking parallel fibers were produced, Purkinje cell dendrites often oriented perpendicular to the thick bundles three-dimensionally. Astrocytes were abundant in all cultures, and there was no definite correlation between the presence of and orientation to Purkinje cell dendrites, although their fibers were frequently associated in parallel with dendrites in the organotypic microexplant culture. Therefore, Purkinje cells may grow their dendrites to the newly produced neurite bundles of parallel fibers in the cerebellar cortex and be oriented at right angles three-dimensionally mainly via "perpendicular contact guidance."
Research Interests: Zoology, Biology, Immunohistochemistry, Medicine, Cerebellum, and 15 moreMice, Animals, Dendrites, Medical Physiology, Glial Fibrillary Acidic Protein, Three Dimensional, Tubulin, Purkinje Cells, Cell communication, Comparative Neurology, Neurosciences, Purkinje cell, Cell count, coculture techniques, and Cell culture techniques
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Research Interests: Chemistry, Medicine, Multidisciplinary, Cerebellum, Low Dose, and 15 morebisphenol A, Mice, Animals, Polychlorinated Biphenyls, Dendrites, Neuronal Development, Nonylphenol, Polychlorinated Biphenyl, Estrogens, Purkinje Cells, Environmental Pollutants, Chemical Structure, Brain Diseases, Purkinje cell, and Thyroid hormone
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Research Interests: Electron Microscopy, HIV, Biological Sciences, In Vitro, Brain, and 15 moreVirus, Animals, Liposomes, Monoclonal Antibodies, Neurons, Astrocytes, Ultrastructure, Rats, Rat, Recombinant Proteins, Enzyme Linked Immunosorbent Assay, Myelin Sheath, Chick embryo, Medical and Health Sciences, and myelin basic protein
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Ten monoclonal antibodies directed against envelope glycoprotein V3 (E) of Japanese encephalitis virus were obtained. They were characterized by hemagglutination inhibition (HI), neutralization, and enzyme-linked immunosorbent assay and... more
Ten monoclonal antibodies directed against envelope glycoprotein V3 (E) of Japanese encephalitis virus were obtained. They were characterized by hemagglutination inhibition (HI), neutralization, and enzyme-linked immunosorbent assay and divided into four types: flavivirus-cross-reactive HI and non-neutralizing antibody (group 1), subgroup-specific HI and non-neutralizing antibody (group 2), low HI and neutralizing antibody (group 3), and non-HI and neutralizing antibody (groups 4 and 5, respectively). Competitive binding assays were performed to analyze the topography of antigenic determinants by enzyme-linked immunosorbent assay. The results of the competitive binding assay separated non-HI and neutralizing antibody into groups 4 and 5, respectively, and demonstrated the existence of at least five distinct antigenic determinants on V3. The site of group 1 was distinct from any other site. The sites of groups 2 and 3 seemed to be located close together. Our results suggest the follo...