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T Hatae

Publications and source records attributed to T Hatae.

At least 19 recordsLinked to original sources
PubMed2001

Prostacyclin-dependent apoptosis mediated by PPAR delta.

Prostacyclin (PGI(2)) plays important roles in hemostasis both as a vasodilator and an endogenous inhibitor of platelet aggregation. PGI(2) functions in these roles through a specific IP receptor, a G protein-coupled receptor linked to G(s) and increases in cAMP. Here, we report that intracellular prostacyclin formed by expressing prostacyclin synthase in human embryonic kidney 293 cells promotes apoptosis by activating endogenous peroxisome proliferator-activated receptor delta (PPAR delta). In contrast, treatment of cells with extracellular prostacyclin or dibutyryl cAMP actually reduced apoptosis. On the contrary, treatment of the cells with RpcAMP (adenosine 3',5'-cyclic monophosphothioate, Rp-isomer), an antagonist of cAMP, enhanced prostacyclin-mediated apoptosis. The expression of an L431A/G434A mutant of PPAR delta completely blocked prostacyclin-mediated PPAR delta activation and apoptosis. These observations indicate that prostacyclin can act through endogenous PPAR delta as a second signaling pathway that controls cell fate.

Amino Acid Sequence↗
PubMed2001

Quasisteady high-confinement reversed shear plasma with large bootstrap current fraction under full noninductive current drive condition in JT-60U.

A quasisteady reversed shear plasma with a large bootstrap current fraction ( approximately 80%) has been obtained for the first time in the JT-60U tokamak. The shrinkage of reversed shear region was suppressed by the bootstrap current peaked at the internal transport barrier (ITB) layer and the ITBs at a large radius were sustained, which, by combination with an H-mode edge pedestal, resulted in a high confinement or 2.2 times the H-mode scaling for 6 times energy confinement time or 2.7 s. Furthermore, a full noninductive current drive was obtained by the bootstrap current and the beam driven current.

Journal Article↗
PubMed2001

Arginine-vasopressin neurons in the rat hypothalamus produce neurokinin B and co-express the tachykinin NK-3 receptor and angiotensin II type 1 receptor.

Secretion of arginine-vasopressin (AVP) from the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei is induced by neurokinin B (NKB) and angiotensin. To characterize the mechanisms by which this occurs, we used immunohistochemical techniques to assess the ability of AVP-producing neurons to express NKB, NKB receptor (NK-3 receptor) and angiotensin II type 1 receptor (AT-1 receptor). Double fluorescence immunohistochemistry indicated that AVP-immunoreactive cell bodies in the PVN and SON, as well as their axon varicosities in the posterior pituitary, co-express NKB. Almost all AVP-neuron perikarya also expressed both the NK-3 receptor and AT-1 receptor. Thus, AVP-producing neurons in the PVN and SON, which are regulated by NKB, are themselves a source of NKB. Furthermore, the regulation of AVP release by these neurons by NKB and angiotensin II is mediated by the NK-3 receptor and the AT-1 receptor, respectively.

Animals↗
PubMed2001

Effects of water deprivation on neurokinin B production by the arginine-vasopressin neurons of hypothalamic paraventricular and supraoptic nuclei.

In order to clarify the role of neurokinin B (NKB), the dynamic changes in NKB expression and synthesis following water deprivation were examined in the arginine-vasopressin (AVP) neurons of hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. In intact rats, NKB and AVP showed almost the same high level of immunohistochemical reactivity in the magnocellular neurons of the PVN and SON, as well as in the varicose fibers in the median eminence (ME). In contrast, NKB precursor peptide (NKBp) immunoreactivity in the SON and PVN were relatively weak. Five days after water deprivation, AVP and NKB immunoreactivity decreased drastically, while NKBp-immunoreactivity increased in both the PVN and SON magnocellular neurons. Reverse transcription-polymerase chain reaction analysis of control animals revealed high levels of AVP mRNA and substantial amounts of NKB mRNA in the SON. This was contrast to the relatively low levels of AVP mRNA and undetectable levels of NKB mRNA in the PVN. After five days of water deprivation, AVP mRNA in the PVN and NKB mRNA in both the PVN and the SON increased considerably. These results indicate that synthesis and release of NKB, which colocalizes to AVP neurons, are enhanced by water deprivation in the same manner as AVP in the PVN and SON. Thus, NKB seems to be involved in the central control of body fluid levels. The results also suggest that the production rate of NKB under normal conditions in SON dominant.

Animals↗
PubMed2000

Actinin-4 is preferentially involved in circular ruffling and macropinocytosis in mouse macrophages: analysis by fluorescence ratio imaging.

We have applied fluorescence ratio imaging to the analysis of an actin-binding protein concentration relative to F-actin in macrophages, in order to explore the role of a novel (alpha)-actinin isoform, actinin-4, relative to that of the classical isoform, actinin-1. Conventional immunofluorescence images showed that both isoforms were enriched in F-actin-rich regions such as cell surface ruffles. However, ratio images further demonstrated that actinin-4 concentrations relative to F-actin were higher in peripheral inward curved ruffles and dorsal circular ruffles, presumed precursor forms of macropinosomes, than in straight linear ruffles, while actinin-1 concentrations were uniform among the different types of ruffles. Macropinosome pulse-labeling and chase experiments indicated that actinin-4 was also closely associated with newly formed macropinosomes and gradually dissociated with their maturation. Consistent with ratio imaging data, macrophages scrape-loaded with anti-actinin-4 showed a more reduced rate of macropinocytosis than those loaded with anti-actinin-1. Altogether, these results indicate that actinin-4 and actinin-1 contribute differently to F-actin dynamics, that actinin-4 is more preferentially involved in early stages of macropinocytosis than actinin-1. A similar redistribution of actinin-4 was also observed during phagocytosis, suggesting that actinin-4 may play the same role in the two mechanistically analogous types of endocytosis, i.e. macropinocytosis and phagocytosis.

Actinin↗
PubMed1999

Fluorescent probe and permeability to cells of isopoly (S-carboxymethyl-L-cysteine) derivative of nucleic acid bases.

Isopoly(S-carboxymethyl-L-cysteine) derivatives of nucleic acid bases were found to form stable complex with oligo-DNA in vitro. Fluorescent probed isopoly(S-carboxymethyl cysteine) derivatives of nucleic acid bases were prepared as antisense oligomers. The transfection of the oligomer into cells was carried out by HVJ-liposome method. Fluorescence was observed from the cells treated with HVJ-liposome including fluorescent probed oligomers.

Animals↗
PubMed1999

Gene transfer of human prostacyclin synthase prevents neointimal formation after carotid balloon injury in rats.

BACKGROUND AND PURPOSE: A disordered proliferative process in the vascular wall is thought to underlie the pathogenesis of restenosis after percutaneous transluminal angioplasty and carotid endarterectomy. A growth inhibitory property of overexpressed prostacyclin (PGI2) synthase (PGIS) was recently implicated in the pathological proliferation of vascular smooth muscle cells (VSMC) in vitro. Here, we investigated the effects of increased PGI2 synthesis on the pathological proliferation of VSMCs. METHODS: The cDNA encoding human PGIS was transfected into endothelium-denuded rat carotid arteries after arterial balloon injury with the use of hemagglutinating virus Japan (HVJ). HVJ liposome vector complex without PGIS cDNA was used for vehicle control. The level of 6-keto PGF1alpha, a stable hydrolyzed metabolite of PGI2, the histological distribution of the immunoreactivity for human PGIS and the ratio of neointimal/medial area were analyzed. RESULTS: In the analyses of 6-keto PGF1alpha, the level in the carotid arteries was significantly elevated 3 days after PGIS expression-vector transfection compared with that in the arteries after vehicle transfection. Seven days after human PGIS expression-vector transfection, the PGIS cDNA-transfected neointimal cells were strongly positive for human PGIS immunoreactivity in 81% sections examined. Fourteen days after the injury, the ratio of neointimal/medial area was 1.2+/-0.4 in the PGIS expression-vector transfected group, which was significantly smaller than that of the vehicle control group, 1.7+/-0.5; P<0.01. CONCLUSIONS: It was thus demonstrated that the gene transfer of human PGIS expression-vector into rat carotid arteries resulted in the increased production of human PGI2 in the vascular wall, the expression of human PGIS in the developing neointima and significantly inhibited the neointimal formation generated after balloon injury.

6-Ketoprostaglandin F1 alpha↗
PubMed1997

The effect of rolipram on the production of cytokines in HTLV-I infected cell lines and peripheral blood mononuclear cells of patients with HTLV-I-associated myelopathy (HAM).

Previous studies have reported that the levels of pro-inflammatory cytokines, such as TNF-alpha and IFN-gamma, are elevated in the serum as well as in the cerebrospinal fluid of HAM/TSP patients. To evaluate the effect of the phosphodiesterase type IV inhibitor, rolipram on cytokine production, peripheral blood mononuclear cells (PBMCs) of HAM/TSP patients or HTLV-I infected T-cell lines (HUT102, MT2) were cultured in the presence of different doses of rolipram. The amount of cytokines in the supernatants of the cultured cells was determined by ELISA for TNF-alpha, IFN-gamma and TGF-beta. Rolipram inhibited TNF-alpha production by HUT102 and PBMCs from all the HAM/TSP patients in a dose-dependent manner. The suppression of IFN-gamma varied and was weaker in some HAM/TSP patients compared to that of TNF-alpha. The concentration of TGF-beta in the culture supernatants was not influenced by rolipram. The levels of TNF-alpha mRNA determined by competitive PCR were not changed in the cultured cells in the presence of rolipram, suggesting that rolipram inhibits TNF-alpha production at the post-transcriptional level. These findings suggest the possible benefit of rolipram as a therapeutic agent for HAM/TSP patients.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1997

Apical tubular network in the rat kidney proximal tubule cells studied by thick-section and scanning electron microscopy.

The apical cytoplasm of several absorbing epithelia contains well-developed apical tubules (AT) which contribute to membrane recycling from endocytic vacuoles to the apical cell membrane. In this study, we examined three-dimensional structures of the AT in rat kidney proximal tubule cells by transmission and scanning electron microscopy. In thin sections, the AT appeared as straight tubules with a rather constant diameter (70-90 nm), but others were curved and, occasionally, branching. No AT were labeled with the marker for the external cell surface (ruthenium red) or exhibited histochemical enzyme activity for lysosomal hydrolase (acid phosphatase). After intravenous injection of horseradish peroxidase, it was absorbed in the kidney proximal tubule cells and the AT were labeled with HRP reaction products. Stereo-viewing of the labeled AT in thick sections revealed that they formed an interconnected tubular network. Scanning electron microscopy allowed a three-dimensional view of the AT, in which a network of branching and anastomosing tubules was revealed. These observations indicate that the AT are intracellular endosomal compartments which form an extensive tubular network in the apical cytoplasm. The possibility that this apical tubular network serves as a large membrane store for membrane recycling is discussed.

Acid Phosphatase↗
PubMed1997

Expression of CD23 in the germinal center of thymus from myasthenia gravis patients.

In order to investigate a pathogenic role of germinal centers which appear in the hyperplastic thymus of myasthenia gravis (MG) patients, we performed an immunohistochemical study using various monoclonal antibodies including CD23. In contrast with tonsilar germinal centers from non-MG individuals, CD23 was strongly and diffusely expressed in the whole area of germinal centers of MG thymi, including the outer zone. In addition, we measured the serum level of soluble CD23 (sCD23) in MG patients at various clinical stages. The high serum sCD23 levels, which were noted in the unthymectomized patients, fell to within normal range over 5 years after thymectomy, and the decline of serum sCD23 correlated well with clinical improvement. CD23 is thought to be responsible for preventing unselected germinal center B cells from entering apoptosis and, in turn, leads to the survival of auto-reactive B cell clones.

Antigens, CD19↗
PubMed1997

The regional distribution and cellular localization of mRNA encoding rat prostacyclin synthase.

The cloned cDNA for rat prostacyclin synthase was found to contain a 1503-bp open reading frame which encoded a 501-amino acid protein sharing 84.0% identity with the human enzyme. RNA blot analysis revealed that the rat prostacyclin synthase mRNA, as a single species of 2.1 kb, is expressed abundantly in the aorta and uterus. High levels of expression were also observed in the stomach, lung, heart, testis, liver, and skeletal muscle. Low but significant expression was also seen in the brain and kidney. Furthermore, the regional distribution and cellular localization of prostacyclin synthase mRNA were examined by in situ hybridization analysis of rat tissue sections. The definitive signals for the mRNA were localized in smooth muscle cells of the arteries, bronchi and uterus, and in the cells of the fibrous tunic surrounding the seminiferous tubules, which are characterized as smooth muscle cells. Besides smooth muscle cells, signals were also detected in the fibroblasts of the heart myocardium, lung parenchyma cells and kidney inner medulla tubules and interstitial cells.

Amino Acid Sequence↗
PubMed1997

Direct measurement of endosomal pH in living cells of the rat yolk sac epithelium by laser confocal microscopy.

Endocytosis of dual-fluorescent dextran by epithelial cells was observed in the perfused rat yolk sac using a confocal laser scanning microscope. Endosomal pH was quantified from confocal images using a dual-fluorescence ratiometry technique. Changes in endosomal pH were followed over time up to 32 min with and without Brefeldin A treatment. After 4 min of endocytosis without Brefeldin A, endosomes had a pH of 6.1 +/- 0.3. After 16 min and 32 min, their pH levels varied widely from 4.0 to 6.6. When Brefeldin A (10 microM) was added to the perfusion medium, endosomal pH remained fairly stable in the range of 6.0 to 6.2 from 4 min to 32 min. The corresponding endosomal structures were examined by electron microscopy after endocytic labeling with horseradish peroxidase. After 4 min, small endocytic vesicles and large endosomal vacuoles with tubular extensions were labeled. After 16 min and 32 min, multivesicular bodies and dense lysosomal structures were progressively labeled, but their labeling was almost undetectable after Brefeldin A treatment. These results suggested that the pH within the sorting compartment of early endosomes is 6.1 +/- 0.3. This is the first quantitative measurement of pH within sorting endosomes in intact cells of the living yolk sac epithelium.

Animals↗
PubMed1996

Antidiuretic action of tachykinin NK-3 receptor in the rat paraventricular nucleus.

Studies were performed on the central antidiuretic actions via the tachykinin NK-3 receptor in the rat hypothalamic paraventricular nucleus (PVN). Microinjections of the selective tachykinin NK-3 receptor agonist senktide (2-200 pmol) into the PVN resulted in prolonged inhibition of urine output in water-loaded rats, its effect being dose-dependent. The antidiuretic action of senktide was blocked by pretreatment with the vasopressin V2 receptor antagonist OPC-31260 (1 mg/kg, i.v.), but not by microinjection of the angiotensin II AT-1 receptor antagonist losartan (1 nmol) into the PVN. NK-3 receptor mRNA was strongly detected in the magnocellular part of the PVN and the supraoptic nucleus (SON) of the hypothalamus as detected by in situ hybridization histochemistry. Moreover, [3H]senktide binding sites were also detected in the PVN and the SON by receptor autoradiography. These findings suggest that NK-3 receptors in the PVN may be involved in water regulation by stimulation of vasopressin secretion from the posterior pituitary gland, and that vasopressin caused water reabsorbtion via the kidney V2 receptor.

Angiotensin Receptor Antagonists↗
PubMed1996

Human gene encoding prostacyclin synthase (PTGIS): genomic organization, chromosomal localization, and promoter activity.

The prostacyclin synthase gene isolated from human genomic libraries (PTGIS) consists of 10 exons spanning approximately 60 kb. All the splice donor and acceptor sites conform to the GT/AG rule. Genomic Southern blot and fluorescence in situ hybridization analyses revealed that the human prostacyclin synthase gene is present as a single copy per haploid genome and is localized on chromosome 20q13. 11-q13.13. The 1.5-kb sequence of the 5'-upstream of the translational initiation site contained both GC-rich and pyrimidine-rich regions and consensus sequences of the transcription factor recognition sites such as Sp1, AP-2, the interferon-gamma response element, GATA, NF-kappaB, the CACCC box, and the glucocorticoid response element. The core binding sequence (GAGACC) of the shear stress responsive element was also found in the 5'-flanking region of the gene. The major product of the primer extension analysis suggested that the transcription of the gene started from the positions around 49 bp upstream of the translational initiation codon. Transient transfection experiments using human aortic and bovine arterial endothelial cells demonstrated that the GC-rich region (positions -145 to -10) possessed a significant promoter activity. The 6-kb downstream sequence of the translational termination codon contained multiple polyadenylation signals, Alu repeat sequences, and the consensus sequence of the primate-repetitive DNA element, MER1. Two sizes of the prostacyclin synthase mRNAs (approximately 6 and 3.3 kb) were detected with the human aorta and lung. RNA blot hybridization analysis using the 3'-untranslated region as probe indicated that the sizes of the 3'-flanking regions were different in the major 6-kb and minor 3.3-kb mRNAs.

Animals↗
PubMed1996

Site-directed mutagenesis of human prostacyclin synthase: Alteration of Cys441 of the Cys-pocket, and Glu347 and Arg350 of the EXXR motif.

The possible active site Cys441 in the Cys-pocket and Glu347 and Arg350 of the EXXR motif of the human prostacyclin synthase, which catalyzes the conversion of prostaglandin H2 to prostacyclin, were subjected to site-directed mutagenesis in order to understand the role of these residues in expressing the enzymatic activity. Five expression vectors encoding the mutant enzymes with a single replacement, Cys441 Ala, Cys441 Ser, Cys441 His, Glu347 Ala and Arg350 Ala, as well as the wild-type enzyme were expressed in 293 cells. The microsomal fraction of the cells expressing the wild-type enzyme showed a specific activity of 96 nmol 6-keto-PGF1alpha/min per mg protein. All of the mutant enzymes examined showed no detectable enzyme activity, although immunoblot analysis demonstrated that levels of all the expressed mutant enzymes were similar to that of the wild-type enzyme. These results indicated that the Cys441 in the Cys-pocket, and Glu347 and Arg350 of the EXXR motif of human prostacyclin synthase are important for expressing the enzymatic activity.

Amino Acid Sequence↗
PubMed1996

Three-dimensional structure of myeloid bodies in the retinal pigment epithelium of the frog, Rana pipiens.

The three-dimensional structures of myeloid bodies (MBs) in the retinal pigment epithelium (RPE) of the frog (Rana pipiens) were studied by transmission and scanning electron microscopy. MBs were observed to be an assemblies of numerous membranous lamellae that were formed by a flattened saccule of the paired membranes in the RPE. On thin-section, they appeared as lenticular, crescentic, circular and spherical forms and were associated with adjacent smooth endoplasmic reticulum (SER) and nuclear membranes. The lenticular MBs were observed to be in connection with the crescentic, circular and spherical forms. Under scanning electron microscopy, the lenticular and crescentic MBs showed a pile of disc-liked lamellae, while the circular and spherical MBs displayed an onion-liked appearance. It was noted that some tubular SER remained in the center of the circular MBs, while the lamellae were compactly arranged and filled the central region in the spherical MBs. All crescentic and spherical MBs were continuous with the lenticular and crescentic MBs. These results suggest that the lenticular MBs may be a precursor form of another variety of MBs which may be transformed into the crescentic, circular and spherical forms by the curving and fusion of the lamellar membranes at the MB's free ends.

Animals↗
PubMed1995

Heritable formation of neuroectodermal tumor in transgenic mice carrying the combined E1 region gene of adenovirus type 12 with the deregulated human renin promoter.

Adenovirus early 1 (E1) region gene products, including E1A and E1B, are required for transcriptional regulation of viral and cellular promoters in infected and transfected culture cells and for transformation of primary rodent cells. Here, we established a line of transgenic mice carrying the E1 region gene of human adenovirus type 12 under the control of the human renin promoter, in which a neuroectodermal tumor derived from retroperitoneal, olfactory, and/or pelvic regions was heritably developed with varying degrees of incidence and the phenotype was successfully passed through six generations. The transgenes were located in the region E2-E3 bands of chromosome 7 with which no genetic linkage to neuroectodermal tumors was previously demonstrated, and expressed only in the tumors but not in another tissue examined. Notably, in addition to the expression of a neural marker gene N-CAM, the three nuclear oncogenes, c-, L-, and N-myc, were coexpressed in the tumors. These results suggest that E1A and E1B are cooperatively involved in the heritable formation of neuroectodermal tumors associated with co-expression of the three sets of myc family genes.

Adenovirus E1 Proteins↗