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T Hatae

Publications and source records attributed to T Hatae.

At least 37 records · Page 2Linked to original sources
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↗
PubMed1994

Neuroectodermal tumors expressing c-, L-, and N-myc in transgenic mice that carry the E1A/E1B gene of human adenovirus type 12.

Adenovirus early 1 (E1) region gene products, including E1A and E1B, are required for transformation of primary cultured rodent cells. In order to investigate in vivo action of the E1 region, we established a line of transgenic mice carrying the oncogenic E1A and E1B genes of human adenovirus type 12 under control of the human angiotensinogen promoter. Histopathological analyses indicated that transgenic mice heritably develop neuroectodermal tumors arising from the pelvic region with varying degrees of incidence. The transgene was expressed in the neuroectodermal tumors as well as in TNT-1 cells, a cell line established from the tumors, where the human angiotensinogen promoter was constitutively active. The high level expression of c-, L-, and N-myc without gene amplification was notable in the original tumors and TNT-1 cells, but not in another tissue examined. The co-expression of the three sets of myc family genes in both the original tumors and the established cell line provided the possibility that the target cells for transformation may belong to a specific cell type that expresses all these oncogenes during development.

3T3 Cells↗
PubMed1994

Detection of human T lymphotrophic virus type I (HTLV-I) proviral DNA and analysis of T cell receptor V beta CDR3 sequences in spinal cord lesions of HTLV-I-associated myelopathy/tropical spastic paraparesis.

Identification of the localization of human T lymphotrophic virus type I (HTLV-I) proviral DNA in the central nervous system (CNS) is crucial to the understanding of the pathogenesis of HTLV-I-associated myelopathy (HAM)/tropical spastic paraparesis (TSP) pathogenesis. We have developed a sensitive detection method, called two-step polymerase chain reaction (PCR) in situ hybridization, which enabled us to detect the HTLV-I proviral DNA in paraffin-embedded spinal cord tissue sections from HAM/TSP patients. HTLV-I proviral DNA was detected only in the nucleus of lymphocytes that had infiltrated into the spinal cord. However, no proviral DNA was amplified in any neuronal cells, including neurons and glial cells. This indicates that the demyelination of the spinal cord by HTLV-I as a result of viral infection of oligodendrocytes or neuronal cells is unlikely. The T cell receptor V beta gene sequence from lymphocytes in the spinal cord lesions taken from the same HAM/TSP autopsy cases revealed unique and restricted CDR3 motifs, CASSLXG(G) (one-letter amino acid. X is any amino acid), CASSPT(G), and CASSGRL which are similar to those described in T cells from brain lesions of multiple sclerosis (MS) and in a rat T cell clone derived from experimental allergic encephalomyelitis (EAE) lesions. The present results suggest that T cells containing restricted V beta CDR3 motifs, which are also found in MS and EAE, become activated upon HTLV-I infection and infiltrate into the spinal cord lesions of HAM/TSP patients.

Adult↗
PubMed1994

Comparative studies on species-specific reactivity between renin and angiotensinogen.

The renin-angiotensin system (RAS) is the most important regulator of electrolyte homeostasis and blood pressure. Our recently generated transgenic mice carrying either the human renin (hREN) or human angiotensinogen (hANG) genes did not develop hypertension but dual gene strains obtained by cross-mating separate lines of mice exhibited a chronically sustained increase in blood pressure, suggesting the presence of species-specific reactivity between renin and angiotensinogen. In order to examine this specificity, the present study was designed to perform a strictly comparative study on hydrolysis of hANG by hREN and mouse submandibular renin (mREN) in vitro by using pure proteins. The recombinant hANG (rhANG) and the synthetic human-type tridecapeptide (hTDP), Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Val-Ile-His, corresponding to the N-terminal sequences of hANG, were used to determine the species specificity of recombinant hREN (rhREN) and mREN. While hTDP was cleaved by both rhREN and mREN with similar Km and with the same order of kcat, rhANG was cleaved by mREN with 16.7-fold higher Km and with 28.2-fold lower kcat than by rhREN. These results showed that kcat/Km value of mREN for rhANG was 468-fold lower than that for rhREN acting on rhANG.

Amino Acid Sequence↗
PubMed1994

Three-dimensional architecture of the tubular endocytic apparatus and paramembranous networks of the endoplasmic reticulum in the rat visceral yolk-sac endoderm.

The three-dimensional architecture of the tubular endocytic apparatus and the endoplasmic reticulum in the rat yolk-sac endoderm was investigated after loading with horseradish peroxidase-conjugated concanavalin A by intrauterine administration. After 30 min, small vesicles (50-150 nm in diameter), small tubules (80-100 nm in diameter) and large vacuoles (0.2-1.0 microns in diameter) in the apical cytoplasm were labeled with the tracer, but lysosomes (1.0-3.5 microns in diameter) in the supranuclear cytoplasm were not labeled until 60 min after loading. Stereo-viewing of the labeled small tubules in thick sections revealed that they were not isolated structures but formed three-dimensional anastomosing networks, which were also confirmed by scanning electron microscopy after maceration with diluted osmium tetroxide. Their earlier labeling with the endocytic tracer, localization in the apical cytoplasm and three-dimensional network formation indicated that the labeled small tubules represented tubular endosomes (tubular endocytic apparatus). These well-developed membranous networks provided by the tubular endosomes are suggested to facilitate the receptor-mediated endocytosis and transcytosis of the maternal immunoglobulin in the rat yolk-sac endoderm. Scanning electron microscopy further revealed lace-like networks of the smooth endoplasmic reticulum near the lateral plasma membrane. Their possible involvement in transport of small molecules or electrolytes is discussed.

Animals↗
PubMed1994

Collagen types in human posterior capsule opacification.

The fibrous type of human posterior capsule opacification was examined by electron microscopy and immunoelectron microscopy to determine which types of collagen were present. The opacification consisted of lens epithelial cells and a large amount of extracellular matrix. The extracellular matrix comprised collagen fibrils and basal lamina-like material. Immunoelectron microscopy revealed that collagen types I, III, and IV were present. Types I and III were localized to the collagen fibrils. Type IV was present in the basal lamina of the lens epithelial cells and in the basal lamina-like material of the extracellular matrix.

Aged↗
PubMed1994

Species-specific expression of the hepatic renin gene.

The catalytic reaction of renin, an aspartyl proteinase, with angiotensinogen is the rate-limiting step fo the renin-angiotensin system involved in the maintenance of blood pressure and electrolyte balance in mammals. We have characterized species-specific expression of the hepatic renin gene by RNase protection experiment, primer extension analysis, and promoter assay using an in vitro DNA transfection. RNase protection experiments revealed that the renin gene is expressed in rat liver, but neither in mouse nor in human. Primer extension analysis identified the putative promoter region of the rat renin gene, which contains TATAAAA sequence, a canonical regulatory DNA element. In order to test whether the upstream region of the renin gene with respect to the putative transcription initiation site is a functional promoter, we have examined the ability of the 5'-flanking sequences of the rat renin gene as well as the human and mouse genes to activate expression of a reporter gene containing the bacterial chloramphenicol acetyltransferase (CAT)-coding sequences, by transient transfection assays. In transfected HepG2 cells, a hepatoma cell line, only the rat renin promoter was capable of driving the CAT gene expression. These results suggested that the rat-specific renin gene expression in the liver could be primarily determined by its promoter specificity.

Animals↗
PubMed1993

Chimeric renin-angiotensin system demonstrates sustained increase in blood pressure of transgenic mice carrying both human renin and human angiotensinogen genes.

A reaction between enzyme renin and its only natural substrate angiotensinogen is the initial and rate-limiting step for producing a potent vasoconstrictor angiotensin II as the final product of the renin-angiotensin system, a contributory factor in the pathogenesis of hypertension. In order to assess the role of the interaction of human renin with human angiotensinogen in the development of high blood pressure, we have constructed the chimeric renin-angiotensin cascade in mice comprising both human renin and human angiotensinogen as well as the endogenous angiotensin-converting enzyme and angiotensin II receptor by cross-mating separate lines of transgenic mice carrying either the human renin or human angiotensinogen genes. Although each single gene carrier did not develop hypertension despite the observed normal tissue-specific expression of the transgenes, dual gene strains exhibited a chronically sustained increase in blood pressure. Administration of a human renin-specific inhibitor (ES-8891) was effective in reducing the elevated blood pressure only against the cross-mated hybrid mice, but treatment of an angiotensin-converting enzyme inhibitor (captopril) and a selective antagonist (DuP 753) directed at the angiotensin II receptor decreased the basal level of blood pressure even in single gene carriers as well as in dual gene mice. These results clearly demonstrated that the sustained increase in blood pressure of the hybrid mice was initiated by the interaction between the products of the two human genes.

Angiotensin I↗
PubMed1993

Rapid freezing replica studies of membrane specializations in the proximal tubules of mouse kidney.

The membrane specializations of the fresh unfixed kidney cortex of adult and neonatal ICR mice were examined by using rapid freezing replica methods. In proximal tubular cells, numerous apical intracellular tubules exhibited helical patterns on the E face with a pitch of about 12 nm. This regular pattern was often continuous with similar striped indentations on the edge of the vacuoles connecting with the tubules. On the luminal surface (ES) of these vacuoles, membrane surface particles were arranged regularly in striped patterns with a center-to-center spacing of about 12 nm. We could not identify differentiations on the PF or PS of the same membrane systems. Another membrane specialization was a plaque or patch of clear pits in tilted lattice alignments on the P face of the large vacuoles with a center-to-center spacing of about 20 nm. This type of specialization was often observed in the neonatal mice proximal tubular cells. These membrane specializations may indicate the active membrane functions in the proximal tubules and suggest the functional continuity and structural relationship of these apical endocytic membrane systems.

Animals↗
PubMed1993

Immunocytochemistry of types I-IV collagen in human anterior subcapsular cataracts.

Human anterior subcapsular cataracts were examined by immunocytochemistry to investigate which types of collagen constitutes the subcapsular fibrous matrix. Types I, III, and IV collagen were shown to be immunologically positive by both light and electron microscopy. In light microscopy, types I and III collagen were stained diffusely whereas type IV collagen was stained in a streaky pattern in the opacities. The lens capsules were stained with antibody to type IV collagen, although the staining patterns were not homogeneous; the inner layers of the capsules were labeled more strongly than the outer layers. Neither type I nor III collagen was detected in the capsules. In electron microscopy, collagen fibrils were labeled with the antibodies to both type I and type III collagen. The lens capsules and multilamellae of the basement membranes in the opacities were labeled with the antibody to type IV collagen.

Basement Membrane↗
PubMed1993

Localization of gap junction proteins, connexins 32 and 26, in rat and guinea pig liver as revealed by quick-freeze, deep-etch immunoelectron microscopy.

By use of site-specific antibodies against synthetic oligopeptides, we examined the localizations of the gap junction proteins connexin 32 (Cx32) and connexin 26 (Cx26) in rat and guinea pig liver. Double-labeling immunofluorescence microscopy revealed that in guinea pig liver both proteins were spread throughout the liver lobules and seemed to localize together within the same gap junction plaque. In rat liver, co-localization of both Cx32 and Cx26 in the same plaques was also suggested in periportal zones. Quick-freeze, deep-etch immunoelectron microscopy showed that immunolabeling of isolated guinea pig liver gap junction plaques with either Cx32 or Cx26 antiserum yielded complete and dense antibody decoration of the cytoplasmic surface of the plaques. In isolated rat liver plaques, the cytoplasmic surfaces were densely decorated with Cx32 antiserum, whereas Cx26 labeling yielded diffuse decoration with variable intensity of the plaques. In both species we did not observe any focal or patchy clusters of the labeling in any plaques examined. Double-labeling immunoelectron microscopy confirmed that both Cx32 and Cx26 are co-localized in the same gap junction plaques. These results suggest that in hepatocytes expressing both Cx32 and Cx26, both types of gap junction proteins are not segregated but intermingle randomly within the same plaques.

Amino Acid Sequence↗
PubMed1993

Dependence of angiotensin production in transgenic mice carrying either the human renin or human angiotensinogen genes on species-specific kinetics of the renin-angiotensin system.

In order to study the functional role of the renin-angiotensin system in the control of blood pressure, projects were initiated using transgenic mice carrying either the human renin gene or the human angiotensinogen gene. To extend the usefulness of the transgenic model system of hypertension research, a two-tiered strategy for generation of transgenic mice with high blood pressure has been developed. This unique system allows the establishment of transgenic lines with a strict species specificity of the renin-angiotensinogen reaction exhibited by transgene products that are regulated in a tissue specific manner. In this report, the strategy concerning the "two-tiered method" is presented and evidence is provided the overproduction of angiotensin occurs only in the combined reaction with human renin and human angiotensinogen expressed in transgenic mice.

Amino Acid Sequence↗
PubMed1993

Cytoplasmic surface ultrastructures of gap junctions in bovine lens fibers.

PURPOSE: To examine the cytoplasmic surface ultrastructures of lens fiber gap junctions, where the cytoplasmic domains of connexons were expected to be exposed. METHODS: Bovine lens fiber gap junctions, both in situ and in the form of isolated membranes, were examined with the deep etching replica methods. Isolated membranes were also examined with the same methods after the treatment with endoproteinase glu-C, which is known to cleave off the cytoplasmic domain of a putative lens fiber connexin MP70 to determine whether any structural changes should occur between proteolyzed and nonproteolyzed gap junctions. In addition, both proteolyzed and nonproteolyzed gap junctions were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunolabeling with the monoclonal antibody that recognized cytoplasmic domain of MP70 to clarify whether MP70 lost its cytoplasmic domain by the treatment with endoproteinase glu-C. RESULTS: Gap junctions were shown to have particulate substructures on their cytoplasmic surfaces; the distributions of the particles were restricted within gap junctional plaques and the non-gap-junctional areas showed smooth cytoplasmic surfaces. Although the treatment with endoproteinase glu-C failed to remove the cytoplasmic particles of gap junctions in deep etching replica study, MP70 was shown to have lost its cytoplasmic domain in sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunolabeling studies. CONCLUSIONS: Each particle revealed on the cytoplasmic surfaces of lens fiber gap junctions corresponded to the cytoplasmic domain of a connexon. The particles were not removed by the treatment with endoproteinase glu-C, whereas MP70 was cleaved by the same treatment.

Animals↗