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N Ueno

Publications and source records attributed to N Ueno.

At least 73 records · Page 4Linked to original sources
PubMed2000

Characterization of follistatin isoforms in early Xenopus embryogenesis.

Follistatin is expressed in Spemann's organizer in the Xenopus gastrula and mimics the activity of the organizer, inducing a neural fate directly in the ectoderm. We have previously shown that follistatin inhibits BMP activity through a direct interaction. In this study, we have characterized the localization and function of two follistatin isoforms to examine the functional differences between them. One notable difference, previously described, is that the shorter form (xFSS or xFS319) but not the C-terminally extended long form (xFSL) associates with cell-surface matrices. Here, we show that the spatial-temporal expression pattern of xFSL and xFSS is indistinguishable. Interestingly, however, xFSS was found to have a more potent inhibitory activity against BMP-4 than xFSL. Furthermore, using a surface plasmon resonance biosensor, xFSS was shown to have a higher binding capacity for BMP subtypes. The diffusion rates of xFSS and xFSL ectopically expressed in Xenopus embryos were similar. Taken together, our results suggest that the difference in BMP-inhibiting activity of the two follistatin isoforms is mainly attributable to a difference in their BMP binding properties rather than to their diffusion rates.

Amino Acid Sequence↗
PubMed1999

Identification of transforming growth factor-beta- regulated genes in caenorhabditis elegans by differential hybridization of arrayed cDNAs.

Members of the transforming growth factor-beta family play critical roles in body patterning, in both vertebrates and invertebrates. One transforming growth factor-beta-related gene, dbl-1, has been shown to regulate body length and male ray patterning in Caenorhabditis elegans. We screened arrayed cDNAs to identify downstream target genes for the DBL-1 signaling by using differential hybridization. C. elegans cDNAs representing 7,584 independent genes were arrayed on a nylon membrane at high density and hybridized with (33)P-labeled DNA probes synthesized from the mRNAs of wild-type, dbl-1, sma-2, and lon-2 worms. Signals for all the spots representing hybridized DNA were quantified and compared among strains. The screening identified 22 and 2 clones, which were positively and negatively regulated, respectively, by the DBL-1 signal. Northern hybridization confirmed the expression profiles of most of the clones, indicating good reliability of the differential hybridization using arrayed cDNAs. In situ hybridization analysis revealed the spatial and temporal expression patterns of each clone and showed that at least four genes, including the gene for the type I receptor for DBL-1, sma-6, were transcriptionally regulated by the DBL-1 signal.

Animals↗
PubMed1999

Isolation and characterization of bone morphogenetic protein-binding proteins from the early Xenopus embryo.

Using a surface plasmon resonance biosensor as a sensitive and specific monitor, we have isolated two distinct bone morphogenetic protein (BMP)-binding proteins, and identified them as lipovitellin 1 and Ep45, respectively. Lipovitellin 1 is an egg yolk protein that is processed from vitellogenin. Both vitellogenin and Ep45 are synthesized under estrogen control in the liver, secreted, and taken up by developing oocytes. In this paper, we have shown that of the TGF-beta family members tested, Ep45 can bind only to BMP-4, whereas lipovitellin 1 can bind to both BMP-4 and activin A. Because of this difference in specificity, we have focused on and further studied Ep45. Kinetic parameters were determined by surface plasmon resonance studies and showed that Ep45 associated rapidly with BMP-4 (k(a) = 1.06 x 10(4) M(-1)s(-1)) and dissociated slowly (k(d) = 1.6 x 10(-4) s(-1)). In Xenopus embryos microinjected with Ep45 mRNA, Ep45 blocked the ability of follistatin to inhibit BMP activity and to induce a secondary body axis in a dose-dependent manner, whereas it had no effect on other BMP antagonists, chordin and noggin. These results support the possibility that Ep45 interacts with BMP to modulate its activities in vivo.

Amino Acid Sequence↗
PubMed1999

A functional bone morphogenetic protein system in the ovary.

Bone morphogenetic proteins (BMPs) comprise a large group of polypeptides in the transforming growth factor beta superfamily with essential physiological functions in morphogenesis and organogenesis in both vertebrates and invertebrates. At present, the role of BMPs in the reproductive system of any species is poorly understood. Here, we have established the existence of a functional BMP system in the ovary, replete with ligand, receptor, and novel cellular functions. In situ hybridization histochemistry identified strong mRNA labeling for BMP-4 and -7 in the theca cells and BMP receptor types IA, IB, and II in the granulosa cells and oocytes of most follicles in ovaries of normal cycling rats. To explore the paracrine function of this BMP system, we examined the effects of recombinant BMP-4 and -7 on FSH (follicle-stimulating hormone)-induced rat granulosa cytodifferentiation in serum-free medium. Both BMP-4 and -7 regulated FSH action in positive and negative ways. Specifically, physiological concentrations of the BMPs enhanced and attenuated the stimulatory action of FSH on estradiol and progesterone production, respectively. These effects were dose- and time-dependent. Furthermore, the BMPs increased granulosa cell sensitivity to FSH. Thus, BMPs have now been identified as molecules that differentially regulate FSH-dependent estradiol and progesterone production in a way that reflects steroidogenesis during the normal estrous cycle. As such, it can be hypothesized that BMPs might be the long-sought "luteinization inhibitor" in Graafian follicles during their growth and development.

Animals↗
PubMed1999

A BMP-inducible gene, dlx5, regulates osteoblast differentiation and mesoderm induction.

Bone morphogenetic proteins (BMPs), members of the transforming growth factor beta superfamily, have been identified by their ability to induce cartilage and bone from nonskeletal cells and have been shown to act as a ventral morphogen in Xenopus mesoderm. We isolated a murine homeobox-containing gene, distal-less 5 (mDlx5), as a BMP-inducible gene in osteoblastic MC3T3-E1 cells. Stable transfectants of MC3T3-E1 that overexpress mDlx5 mRNA showed increase in various osteogenic markers, a fourfold increase in alkaline phosphatase activity, a sixfold increase in osteocalcin production, and appearance in mineralization of extracellular matrix. Furthermore, mDlx5 was induced orthotopically in mouse embryos treated with BMP-4 and in fractured bone of adult mice. Consistent with these observations, we also found that injection of mDlx5 mRNA into dorsal blastomeres enhanced the ventralization of Xenopus embryos. These findings suggest that mDlx5 is a target gene of the BMP signaling pathway and acts as an important regulator of both osteogenesis and dorsoventral patterning of embryonic axis.

3T3 Cells↗
PubMed1999

Negative regulation of Wingless signaling by D-axin, a Drosophila homolog of axin.

Wnt/Wingless directs many cell fates during development. Wnt/Wingless signaling increases the amount of beta-catenin/Armadillo, which in turn activates gene transcription. Here the Drosophila protein D-Axin was shown to interact with Armadillo and D-APC. Mutation of d-axin resulted in the accumulation of cytoplasmic Armadillo and one of the Wingless target gene products, Distal-less. Ectopic expression of d-axin inhibited Wingless signaling. Hence, D-Axin negatively regulates Wingless signaling by down-regulating the level of Armadillo. These results establish the importance of the Axin family of proteins in Wnt/Wingless signaling in Drosophila.

Adaptor Proteins, Signal Transducing↗
PubMed1999

XIAP, a cellular member of the inhibitor of apoptosis protein family, links the receptors to TAB1-TAK1 in the BMP signaling pathway.

Signals elicited by transforming growth factor-beta (TGF-beta) superfamily ligands are generated following the formation of heteromeric receptor complexes consisting of type I and type II receptors. TAK1, a member of the MAP kinase kinase kinase family, and its activator, TAB1, participate in the bone morphogenetic protein (BMP) signaling pathway involved in mesoderm induction and patterning in early Xenopus embryos. However, the events leading from receptor activation to TAK1 activation remain to be identified. A yeast interaction screen was used to search for proteins that function in the pathway linking the receptors and TAB1-TAK1. The human X-chromosome-linked inhibitor of apoptosis protein (XIAP) was isolated as a TAB1-binding protein. XIAP associated not only with TAB1 but also with the BMP receptors in mammalian cells. Injection of XIAP mRNA into dorsal blastomeres enhanced the ventralization of Xenopus embryos in a TAB1-TAK1-dependent manner. Furthermore, a truncated form of XIAP lacking the TAB1-binding domain partially blocked the expression of ventral mesodermal marker genes induced by a constitutively active BMP type I receptor. These results suggest that XIAP participates in the BMP signaling pathway as a positive regulator linking the BMP receptors and TAB1-TAK1.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Decreased food intake and body weight in pancreatic polypeptide-overexpressing mice.

BACKGROUND & AIMS: Pancreatic polypeptide (PP) is a 36-amino acid hormone produced by F cells within the pancreatic islets and the exocrine pancreas. The definitive function of PP in mammalian physiology remains to be determined. This study examined the effects of chronic overexpression of PP through the development of PP transgenic mice. METHODS: PP transgenic mice were created by using mouse PP complementary DNA under the control of the cytomegalovirus immediate early enhancer-chicken beta-actin hybrid promoter (pCAGGS expression vector). RESULTS: A unique line of transgenic mice was created that overexpresses PP in the pancreatic islets with low levels of expression in other tissues including the brain. Plasma PP concentrations were more than 20 times higher than those of control littermates. However, PP overproduction led to postnatal lethality in half of the pups because of markedly decreased milk intake. The remaining PP transgenic mice gained less weight with specifically reduced food intake and fat mass compared with controls, a result that was more evident in male than in female mice. The transgenic mice exhibited a reduced rate of gastric emptying of a solid meal but had normal oxygen consumption and fasting leptin levels. Immunoneutralization with anti-PP antiserum reversed the phenotypic changes of transgenic animals. CONCLUSIONS: PP could be involved in feeding and body weight regulation partly through regulation of gastric emptying.

Animals↗
PubMed1999

Urocortin reduces food intake and gastric emptying in lean and ob/ob obese mice.

BACKGROUND & AIMS: Gastric emptying plays an important role in regulating food intake. This study was designed to investigate whether intraperitoneally injected urocortin reduces gastric emptying, feeding, and body weight in lean and ob/ob obese mice. METHODS: Food intake and body weight were measured after intraperitoneal injections of one of the following: urocortin, deamidated form of urocortin (urocortin OH), corticotropin-releasing factor (CRF), CRF6-33, cholecystokinin octapeptide (CCK-8), and leptin in 16-hour food-deprived animals. Gastric emptying was assessed 2, 4, or 8 hours after intraperitoneal injection. Repeated injections of urocortin were continued for 5 days in ob/ob mice. RESULTS: Urocortin (0.003-3 nmol) dose-dependently and potently decreased food intake and body weight gain in lean mice. The ranking order of potency was urocortin > urocortin OH >/= CRF > CCK-8 > CRF6-33 > leptin. Gastric emptying was also potently reduced by urocortin with a similar ranking order of potency of urocortin > CRF > urocortin OH > CCK-8. Simultaneous administration of urocortin and CRF receptor antagonist, alpha-helical CRF9-41, blocked the effects of urocortin. Urocortin reduced food intake and body weight gain, as well as the rate of gastric emptying, in ob/ob mice, which was significantly faster than that of lean mice. Five daily injections of urocortin significantly lowered body weight and improved glycemic control in ob/ob mice. CONCLUSIONS: The urocortin-induced decrease in food intake and body weight in lean and ob/ob mice is closely related to gastric emptying and opens new possibilities for the treatment of obesity.

Animals↗
PubMed1999

Assessment of residual bile duct stones with use of intraductal US during endoscopic balloon sphincteroplasty: comparison with balloon cholangiography.

BACKGROUND: We sought to determine the utility of intraductal ultrasonography (IDUS) in detecting residual bile duct stones during endoscopic balloon sphincteroplasty. METHODS: Eighty-one consecutive patients with bile duct stones who underwent IDUS during endoscopic balloon sphincteroplasty were studied. IDUS was performed with a thin-caliber ultrasonic probe (diameter 2.0 mm, frequency 20 MHz) by a transpapillary route after stone extraction. When IDUS or balloon-retrograde cholangiography suggested residual stones, the bile duct was cleared again with a Dormia basket. Extraction of the stones was confirmed by direct duodenoscopic visualization. Videotapes of IDUS and cholangiograms were reviewed retrospectively without knowledge of the results of other diagnostic modalities. RESULTS: In 27 of 81 patients (33%), IDUS detected small residual stones not seen on cholangiography. When stones were fragmented with mechanical lithotripsy, the accuracy of IDUS in detecting small residual stones was significantly greater than that of balloon-endoscopic retrograde cholangiography (95% vs 50%, p < 0. 001). When the bile duct was greater than 10 mm in diameter, the accuracy of IDUS in detecting small residual stones was significantly greater than that of cholangiography (92% vs. 56%, p < 0.001). CONCLUSIONS: IDUS is useful for detecting small residual bile duct stones during endoscopic balloon sphincteroplasty when stones are fragmented by mechanical lithotripsy or when there is evidence of a dilated bile duct (>10 mm).

Adult↗
PubMed1999

Pancreatitis induced by endoscopic balloon sphincter dilation and changes in serum amylase levels after the procedure.

BACKGROUND: The short-term safety of endoscopic balloon sphincter dilation remains to be defined. We prospectively investigated the rate of pancreatitis and changes in pancreatic enzyme values induced by endoscopic balloon sphincter dilation. METHODS: One hundred nine patients with bile duct stones were treated using endoscopic balloon sphincter dilation between July 1995 and September 1997. Serum amylase values were determined before and after the procedure in 100 patients treated 106 times, and these values were compared with the incidence of pancreatitis. Patients were classified into 3 categories according to serum amylase levels before therapy (group A, within the normal range; group B, above the normal upper limit up to 500 mU/mL; group C, above 500 mU/mL). The severity of pancreatitis was categorized into 4 grades (minimal, postprocedural abdominal pain lasting no longer than the next morning; mild, elevation of amylase values to at least three times normal accompanied by postprocedural abdominal pain necessitating treatment for 3 nights; moderate, required 4 to 10 nights of treatment; severe, required intensive care). RESULTS: Postprocedural serum amylase levels in groups A and B were significantly elevated compared with baseline levels. In contrast, these levels in group C were decreased. Overall, postprocedural amylase levels were elevated in 95 studies. Minimal or mild pancreatitis developed in 21 (15 minimal and 6 mild) patients in groups A and B only. CONCLUSIONS: Although pancreatic serum enzymes were elevated in almost all patients after endoscopic balloon sphincter dilation, procedure-related pancreatitis developed in a small percentage. There does not seem to be any contraindication to the use of endoscopic balloon sphincter dilation in patients with high amylase levels and an indication for the procedure, although such patients sometimes have stone-related pancreatitis or may be predisposed to developing pancreatitis.

Aged↗
PubMed1999

Treatment of gastric tumors by endoscopic mucosal resection with a ligating device.

BACKGROUND: This study attempted to determine the indication for endoscopic mucosal resection with a ligating device (EMRL) and to assess the efficacy of radical (complete) resection of early gastric carcinoma and adenoma. METHODS: Sixteen patients with early gastric carcinoma (17 lesions) and 21 patients with gastric adenoma (23 lesions) underwent EMRL with an endoscope with a ligating device. After epinephrine solution was injected into the submucosa, the lesions were aspirated, ligated, and resected. RESULTS: Twelve of 17 early carcinomas (70.6%) and 18 of 23 adenomas (78.3%) were radically resected by EMRL. The average size of the resected specimens was 12.8 x 11.0 mm. The rate of successful radical resection by EMRL, including piecemeal resection, was 100% (15/15) for lesions located in the antrum, 80% (4/5) in the angle, 61.1% (11/18) in the body, and 0% (0/2) for lesions at the cardia. Repeat EMRL was performed successfully in cases of partial resection (n = 3). No serious complication was encountered. No recurrence of the tumors was identified in cases of radical resection during a median follow-up period of 22.8 months. CONCLUSION: EMRL is suitable for the treatment of gastric tumorous lesions. For the treatment of early carcinoma, well-differentiated mucosal carcinomas smaller than 10 mm located in the distal stomach represent the best indication for EMRL.

Adenocarcinoma↗
PubMed1999

Mouse pancreatic polypeptide modulates food intake, while not influencing anxiety in mice.

This study was designed to investigate the effects of synthetic mouse pancreatic polypeptide (mPP) on feeding and anxiety in mice. The intracerebroventricular (i.c.v.) injection of mPP (0.003-3 nmol) dose-dependently increased food intake. A significant increase was observed 20 min after i.c.v. injection and continued for 4 h. The intraperitoneal (i.p.) injection of mPP (0.03-30 nmol) dose-dependently decreased food intake. A significant decrease was observed 20 min after i.p. injection and continued for 4 h. In the elevated plus maze test, the i.c.v. injection of mPP (0.003-3 nmol) did not affect anxiety behavior. These results suggest that mPP modulates food intake and the Y4 receptor in the brain may contribute to the regulation of feeding, whereas appearing not to influence anxiety in mice.

Animals↗
PubMed1999

Restricted expression of the receptor serine/threonine kinase BMPR-IB in zebrafish.

Bone morphogenetic proteins (BMPs) comprise a rapidly expanding subclass of the transforming growth factor-beta superfamily. They are known to regulate a diverse range of developmental phenomena including cell differentiation, morphogenesis and apoptosis. In this study, we have isolated a zebrafish homolog of BMP type IB receptor (BMPR-IB) and examined the localization of the transcripts during embryogenesis. Whole-mount in situ hybridization analysis revealed that unlike other type I and type II receptors that mediate BMP signal, it is expressed in developing somite and in mid-hind brain region in a restricted manner.

Amino Acid Sequence↗
PubMed1999

Differential response of macrophage subpopulations to myelin degradation in the injured rat sciatic nerve.

Molecular mechanisms of myelin removal by macrophages were explored by examining the immunophenotypes of macrophages following injury of rat sciatic nerve, using a combined method of immunohistochemistry and confocal laser microscopy. In the crush injury model, the involvement in myelin clearance of a cytoplasmic antigen specific for monocytes/macrophages, ED1, was evident. The obvious recruitment of ED1-immunoreactive (-ir) cells was detected first at the crush injury site and then in the distal stump within which Wallerian degeneration had occurred. Double labelling revealed that the ED1-ir cells, except for monocyte-like round cells, always phagocytosed myelin basic protein-ir myelin debris. On the other hand, the expression of ED2, a surface antigen specific for resident macrophages, was significantly different; ED2-ir cells also increased while myelin removal was progressing from day 3 to day 7, but only some of the cells were engaged in myelin phagocytosis. The poor capacity of myelin phagocytosis by ED2-ir cells was supported by the transection model, in which the proximal stump was ligated to suppress regeneration. ED2 may be involved in events other than myelin removal, providing a local environment conducive to axonal regeneration. Our findings thus seem to suggest that ED1 is one of the most reliable markers for cells carrying out myelin phagocytosis, whereas ED2 may participate in entirely different functions. The expression of complement receptor type 3, OX42, was similar to that of ED1 in terms of the swift recruitment of immunopositive cells, their distribution with close association to myelin debris and their high phagocytotic capacity. This supports previously reported in vitro evidence that myelin phagocytosis by macrophages may be complement-mediated.

Animals↗
PubMed1999

Identification and characterization of E-APC, a novel Drosophila homologue of the tumour suppressor APC.

BACKGROUND: Mutations in the adenomatous polyposis coli (APC) tumour suppressor gene are implicated in the genesis of colorectal cancers. The product of the APC gene forms a complex with beta-catenin, glycogen synthase kinase 3beta (GSK-3beta) and Axin/conductin, and induces the degradation of beta-catenin. RESULTS: We have identified a novel Drosophila homologue of APC, E-APC, which is similar to but differs in several respects from D-APC. The E-APC cDNA encodes a protein of predicted 1067 amino acids, with seven armadillo repeats, two copies of the 15-amino acid repeat, five copies of the 20-amino acid repeat, and one Axin/conductin binding site. E-APC directly interacts with D-Axin and Armadillo (Arm, the Drosophila homologue of beta-catenin) in vitro, destabilizes intracellular beta-catenin, and suppresses beta-catenin/TCF-regulated transcription in APC-/- colon cancer cells. The E-APC mRNA is ubiquitously expressed throughout all developmental stages in Drosophila. CONCLUSION: Our findings suggest that E-APC may be universally involved in the regulation of the Wingless signalling pathway by down-regulating the level of Arm in Drosophila.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Smad8B, a Smad8 splice variant lacking the SSXS site that inhibits Smad8-mediated signalling.

BACKGROUND: Members of the TGF-beta superfamily of ligands bind to and activate surface serine/threonine-kinase receptors. Transduction of these signals requires the Smad proteins, which transiently interact with the activated receptor complex and are phosphorylated on their C terminus, SSXS site, by the type I receptor. Smad8 is a downstream signalling mediator of ALK2/ActRIA. RESULTS: We have cloned a splice variant of Smad8, designated Smad8B. The Smad8 and Smad8B cDNAs are identical in sequence, except that Smad8B lacks a portion encoding 47 amino acids, including the SSXS phosphorylation site, in the C-terminal MH2 region. Both Smad8 and Smad8B were expressed in many of the same cell types. Smad8B was capable of specific complex formation with either Smad8 or Smad4 in mammalian cells. In cells expressing constitutively activated ALK2, Smad8B was localized to the cytoplasmic region, whereas Smad8 was translocated into the nucleus. In mammalian cells, Smad8B acted as a dominant inhibitor of BMP signalling. CONCLUSIONS: Smad8B, a splice variant of Smad8, was isolated and found to specifically associate with both Smad8 and Smad4. Smad8B inhibited BMP signalling. Smad8 and Smad8B thus represent novel signal transduction proteins that may regulate the BMP signalling pathway.

Alternative Splicing↗