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Y Nabeshima

Publications and source records attributed to Y Nabeshima.

At least 73 records · Page 4Linked to original sources
PubMed1996

Promoter structure of the mouse TATA-binding protein (TBP) gene.

5'-RACE and genomic cloning were used to determine that the mouse TBP (mTBP) gene consists of one 5'-terminal non-coding exon followed by seven protein-coding exons. The region upstream of the first exon lacks a TATA-box. Hence, as with the case of other genes carrying TATA-less promoter, transcription starts from a cluster of sites which are located at the restricted region of mTBP gene. Interestingly, sequences of this region are well conserved between human and mouse TBP genes, suggesting that both mouse and human TBP genes drive mRNA synthesis in a similar way, and the sequence homology between two genes was used to assign a putative start point for the human TBP gene. Mouse, human, and Trimersurus gramineus (green habu snake) TBP genes share two GC-rich regions in their promoter regions. Thus, it is probable that diverse species of vertebrates commonly use TATA-less promoter bearing GC-rich regions to direct ubiquitous TBP expression.

Amino Acid Sequence↗
PubMed1996

Uniaxial tension inhibits tendon collagen degradation by collagenase in vitro.

Tendon structure is governed largely by factors regulating the anabolic and catabolic phases of tenocyte metabolism. Little is known about the mechanisms that regulate the synthesis, activation, and action of metalloproteinases, which are key enzymes in a multifactorial cascade controlling homeostasis of the extracellular matrix. In the present study, we investigated the effect of tension on collagenase-induced degradation of the tendon in vitro by assessing changes in structural and material properties measured during tensile failure tests. Devitalized right-left pairs of rabbit patella-patellar tendon-tibia units were maintained under culture conditions in the presence of 60 U/ml highly purified collagenase for 20 hours. One randomly selected unit from each animal was subjected to a tension that produced a constant 4% elongation or strain (n = 10); the contralateral unit served as a slack comparison (n = 10). In one series of experiments (immediate, n = 5), the tension was applied immediately prior to collagenase exposure. In a second series (delayed, n = 5), it was delayed for 4 hours to allow time for the collagenase to diffuse into the tendon. Additional devitalized and nonincubated units (n = 6) were used as normal controls. Collagenase exposure caused large decreases in stiffness and elongation to failure in slack units. This resulted in greater than 80% reductions in both maximum failure force and energy to failure. In contrast, the loaded unit in both experimental protocols had significantly greater stiffness than control units. In both the immediate and the delayed protocols, the loaded tendons had significantly higher stiffness and failed at significantly higher elongations and maximum forces than the slack tendons. Diffusion studies with and without tension showed the tension did not inhibit diffusion of collagenase into the tendon but did significantly decrease the water content from 64.6 to 57.8%. The data suggest that stresses and strains of the extracellular matrix may modify the kinetics of the bacterial collagenase-collagen interaction. Matrix stress and strain may be an important and overlooked factor that modulates the susceptibility of collagen to proteolytic degradation.

Animals↗
PubMed1996

Lysophosphatidic acid and bFGF control different modes in proliferating myoblasts.

Myogenic cells provide excellent in vitro models for studying the cell growth and differentiation. In this study we report that lysophosphatidic acid (LPA), a bioactive phospholipid contained in serum, stimulates the growth and inhibits the differentiation of mouse C2C12 myoblast cells, in a distinct manner from basic fibroblast growth factor (bFGF) whose mitotic and anti-differentiation actions have been well investigated. These actions of LPA were both blocked by pertussis toxin, suggesting the involvement of Gi class of G proteins, whereas bFGF acts through receptor tyrosine kinases. Detailed analysis revealed that LPA and bFGF act differently in regulating the myogenic basic helix-loop-helix (bHLH) proteins, the key players in myogenic differentiation process. LPA stimulates the proliferation of undifferentiated myoblasts allowing the continued expression of MyoD, but in contrast, bFGF does so with the MyoD expression suppressed at the mRNA level. Both compounds maintain the myf-5 expression, and suppress the myogenin expression. In addition, while LPA did not inhibit cell-cell contact-induced differentiation, bFGF strongly inhibited this process. Furthermore, LPA and bFGF act cooperatively in their mitogenic and anti-differentiation abilities. These findings indicate that LPA and bFGF differently stimulate intracellular signaling pathways, resulting in proliferating myoblasts each bearing a distinct expression pattern of myogenic bHLH proteins and distinct differentiation potentials in response to cell-cell contact, and illustrate the biological significance of Gi-mediated and tyrosine kinase-mediated signals.

Animals↗
PubMed1996

Hikaru genki protein is secreted into synaptic clefts from an early stage of synapse formation in Drosophila.

The development of neural circuits is regulated by a large number of factors that are localized at distinct neural sites. We report here the localization of one of these factors, hikaru genki (hig) protein, at synaptic clefts in the pupal and adult nervous systems of Drosophila. In hig mutants, unusually frequent bursting activity of the muscles and abnormal motor behavior during the adult stage suggest the misfunction of neuromuscular circuitry. Our immunohistochemical analyses revealed that hig protein, produced by neurons, is secreted from the presynaptic terminals into the spaces between the presynaptic and postsynaptic terminals. In addition, we have found that the localization of this protein in the synaptic spaces temporally correlates with its functional requirement during a critical period that occurs in the middle stage of pupal formation, a period when a number of dendrite and axon growth cones meet to form synapses. These findings indicate that hig protein functions in the formation of functional neural circuits from the early stages of synapse formation.

Aging↗
PubMed1995

Asymmetric segregation of the homeodomain protein Prospero during Drosophila development.

Asymmetric divisions that produce two distinct cells play fundamental roles in generating different cell types during development. In the Drosophila central nervous system, neural stem cells called neuroblasts divide unequally into another neuroblast and a ganglion mother cell which is subsequently cleaved into neurons. Correct gene expression of ganglion mother cells requires the transcription factor Prospero. Here we demonstrate the asymmetric segregation of Prospero on neuroblast division. Prospero synthesized in neuroblasts is retained in the cytoplasm and at mitosis is exclusively partitioned to ganglion mother cells, in which it is translocated to the nucleus. Differential segregation of Prospero was also found in the endoderm. We have identified a region in Prospero that is responsible for this event. The region shares a common motif with Numb, which also shows unequal segregation. We propose that asymmetric segregation of transcription factors is an intrinsic mechanism for establishing asymmetry in gene expression between sibling cells.

Amino Acid Sequence↗
PubMed1995

A metalloprotease-disintegrin participating in myoblast fusion.

Skeletal muscle development involves the formation of multi-nucleated myotubes. This is thought to proceed by the induction of differentiation (acquisition of fusion competence) of myoblast cells, their aggregation, and union of their plasma membranes. Various membrane proteins including N- and M-cadherins, N- and V-CAMs and integrins participate in myotube formation, but the molecular mechanisms of muscle cell fusion are poorly understood. Here we report the identification of three new, myoblast-expressed gene products, meltrin-alpha, beta and gamma, with homology to both viper haemorrhagic factors and fertilin (PH-30), a membrane protein involved in egg-sperm fusion. Meltrin-alpha, a member of the metalloproteinase/disintegrin protein family, appears to be required for myotube formation. Involvement of a fertilin-related protein in myogenesis suggests that there are common mechanisms in gamete and myoblast fusion.

ADAM Proteins↗
PubMed1995

Exercise at high temperature causes maternal hyperthermia and fetal anomalies in rats.

Hyperthermia is thought to be a teratogen in many animal species and also in humans. It has been reported that hyperthermia caused by sauna, hot tub, or fever during the early stages of pregnancy is related to an increased risk for neural tube defects. During exercise, especially in hot conditions, body temperature can also rise to fairly high levels. Thus, we can surmise that hyperthermia induced by exercise can also cause fetal malformation. To investigate this hypothesis, pregnant rats at 9 days of gestation were divided into four groups. In the first group, the animals were made to swim for 30 minutes in water at a temperature of 40.5 degrees C. In the second group, they were restrained and immersed in water for the same time at the same temperature. In the third group, the rats were forced to swim in water at 36.0 degrees C. The fourth group were controls. The core temperature of the rats was measured during these procedures. On the 18th gestational day, fetuses were extracted by cesarean section. The elevation of maternal core temperature was significantly greater in the first group than in the other groups. In the first group, 69% of fetuses had various external anomalies. No anomalies were found in the other groups. Our results show that exercise in hot conditions caused the elevation of core temperature and resulted in fetal anomalies in rats.

Animals↗
PubMed1995

Effect of fibrin glue and endothelial cell growth factor on the early healing response of the transplanted allogenic meniscus: a pilot study.

Twentyfour meniscal allotransplantations were conducted in 12 adult mongrel dogs. The medial meniscus was replaced using a deep-frozen meniscal allograft. The junction between the meniscus and capsule was treated in one of the three ways. In the control group, the meniscus was sutured only to the adjacent capsular tissue (group C). In the second group, fibrin glue was injected at the junction (group F), and in the third group, fibrin glue and endothelial cell growth factor (ECGF) were injected at the juncture between the transplanted meniscus and the adjacent capsule before the meniscus was sutured (group FE). Histological observation was performed to investigate the effect of fibrin glue and ECGF on the healing process of transplants at various intervals of 1, 4, 8 and 12 weeks. No immunological response was noted in any of the knees. The healing of the transplanted meniscus was first observed at the peripheral attachment. Also, the pannus-like tissue extended from the synovium to the surface of the meniscus. The healing rate in each group at 1 week and 12 weeks was 22% and 77% in group C, 52% and 80% in group F, and 64% and 80% in group FE, respectively. At 4 and 8 weeks, early cellular repopulation was found in group FE and the area which contained new cells was larger than the acellular central core at 8 weeks. However, there was no difference among the three groups at 12 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

The delta-crystallin enhancer-binding protein delta EF1 is a repressor of E2-box-mediated gene activation.

The repressor delta EF1 was discovered by its action on the DC5 fragment of the lens-specific delta 1-crystallin enhancer. C-proximal zinc fingers of delta EF1 were found responsible for binding to the DC5 fragment and had specificity to CACCT as revealed by selection of high-affinity binding sequences from a random oligonucleotide pool. CACCT is present not only in DC5 but also in the E2 box (CACCTG) elements which are the binding sites of various basic helix-loop-helix activators and also the target of an unidentified repressor, raising the possibility that delta EF1 accounts for the E2 box repressor activity. delta EF1 competed with E47 for binding to an E2 box sequence in vitro. In lymphoid cells, endogenous delta EF1 activity as a repressor was detectable, and exogenous delta EF1 repressed immunoglobulin kappa enhancer by binding to the kappa E2 site. Moreover, delta EF1 repressed MyoD-dependent activation of the muscle creatine kinase enhancer and MyoD-induced myogenesis of 10T1/2 cells. Thus, delta EF1 counteracts basic helix-loop-helix activators through binding site competition and fulfills the conditions of the E2 box repressor. In embryonic tissues, the most prominent site of delta EF1 expression is the myotome. Myotomal expression as well as the above results argues for a significant contribution of delta EF1 in regulation of embryonic myogenesis through the modulation of the actions of MyoD family proteins.

Base Sequence↗
PubMed1993

Myogenin gene disruption results in perinatal lethality because of severe muscle defect.

Myogenin is a member of the basic helix-loop-helix (bHLH) gene family and converts multipotential mesodermal cells to myoblasts. The four members of the myoD family show unique spatio-temporal expression patterns and therefore may have different functions during myogenesis. Here we inactivate the myogenin gene in order to understand its role in myogenesis. Homozygous mutations are lethal perinatally owing to the resulting major defects in skeletal muscle. The extent of disorganization of muscle tissue differs in three regions. In the latero-ventral body wall, most cells, including myogenic cells, disappear and there is rapid accretion of fluid. In the limbs, cells of the myogenic lineage exist, but they are severely disrupted, and some of them are mono-nucleate with properties of myoblasts. In contrast, there are many axial, intercostal and back muscle fibres to be seen, although fibres are mainly disorganized and Z-lines are not present in most myofibrils. These findings are evidence that myogenin is crucial for muscle development in utero and demonstrate that other members of the myogenic gene family cannot compensate for the defect.

Animals↗
PubMed1993

Expression of myogenic factors in denervated chicken breast muscle: isolation of the chicken Myf5 gene.

In this study, we have isolated and characterized the chicken Myf5 gene, and cDNA clones encoding chicken MyoD1 and myogenin. The chicken Myf5 and MRF4 genes are tandemly located on a single genomic DNA fragment, and the chicken Myf5 gene is organized into at least three exons. Using genomic and cDNA probes, we further analyzed the mRNA levels of four myogenic factors during chicken breast muscle development. This analysis revealed that myogenin expression is restricted to in ovo stages in breast muscle, and is not detectable in neonatal and adult stages. On the other hand, Myf5 expression is detectable until day 7 post-hatching, and is not found in adult muscle, whereas high levels of MyoD1 and MRF4 are detectable at all stages. To further understand the roles of innervation on muscle maturation, we analyzed the expression of the four myogenic factors in denervated adult breast muscle. We found that MyoD1, myogenin, and MRF4 are induced at high levels in denervated muscle, whereas no change occurs in the level of Myf5. These studies suggest that innervation controls the relative abundance and type of myogenic factors that are expressed in adult muscle, and that when nerve control is removed, the muscle reverts to a neonatal phenotype, with the enhanced expression of three myogenic factors (MyoD1, myogenin, and MRF4).

Amino Acid Sequence↗
PubMed1993

Upstream region of the myogenin gene confers transcriptional activation in muscle cell lineages during mouse embryogenesis.

Myogenin, one of the MyoD-related factors containing basic-helix-loop-helix motifs, is transcriptionally activated in skeletal muscle lineages in somites and limb buds during embryogenesis. In an attempt to understand regulatory mechanisms which govern transcriptional activation of the myogenin gene, transgenic mice bearing the lacZ gene driven by the upstream region of the myogenin gene were generated. Stereoscopic visualization of LacZ-positive cells of these transgenic mouse embryos revealed that the upstream region of the myogenin gene conferred its transcriptional activation in cells of the skeletal muscle lineages in somites, limb buds, and visceral arches. Moreover, transient LacZ expression in newly formed somites in addition to its strong activation in myotomal regions of mature somites with a rostro-caudal gradient raised the possibility that myogenin is transcriptionally activated in immature somites before myotome formation.

Animals↗
PubMed1993

hikaru genki, a CNS-specific gene identified by abnormal locomotion in Drosophila, encodes a novel type of protein.

We have identified a gene, hikaru genki (hig), whose mutant phenotype includes abnormal locomotor behavior. Mutant first instar larvae have uncoordinated movements, and both larvae and adults have reduced locomotion. Sequence analyses revealed that this gene encodes a novel type of protein with a signal sequence, but without transmembrane regions. One of its domains has similarities with immunoglobulin domains; three or four regions are similar to a complement-binding domain found in complement-related proteins and selectins. In situ hybridization to embryos revealed that accumulation of the hig transcripts is restricted to subsets of cells in the CNS. Our data suggest that hig has a role in the development of CNS functions involved in locomotor activity.

Amino Acid Sequence↗