Biomedical subjects

S Yuasa

Publications and source records attributed to S Yuasa.

At least 37 records · Page 2Linked to original sources

PubMed2001

Testicular germ cell apoptosis in Bcl6-deficient mice.

Bcl6 protein has been detected in testicular germ cells, mainly spermatocytes, of normal mice, but its physiological role is largely unknown. The number of spermatozoa in the cauda epididymis of adult Bcl6-deficient (Bcl6-/-) mice is lower than that of Bcl6+/+ mice. We have found numerous apoptotic spermatocytes at the metaphase I stage with induction of Bax protein in adult Bcl6-/- testes. Developmentally, the incidence of germ cell apoptosis of Bcl6-/- mice was similar to that of Bcl6+/+ mice until six weeks of age and increased after eight weeks of age. The incidence of apoptosis in heterozygous Bcl6+/- mice was also higher than that of Bcl6+/+ mice. Since the activated form of p38 MAP kinase was detected in spermatocytes of adult Bcl6-/- mice, the germ cell apoptosis may be induced by stressors. Treatment of testes of adult Bcl6+/+ mice with a mild hyperthermia resulted in germ cell apoptosis predominantly in metaphase I spermatocytes with induction of Bax protein and activation of p38 MAP kinase and this apoptosis mimics that in adult Bcl6-/- mice. Thus, Bcl6 may play a role as a stabilizer in protecting spermatocytes from apoptosis induced by stressors.

Animals↗

PubMed2001

The role of presenilin 1 during somite segmentation.

The Notch signalling pathway plays essential roles during the specification of the rostral and caudal somite halves and subsequent segmentation of the paraxial mesoderm. We have re-investigated the role of presenilin 1 (Ps1; encoded by Psen1) during segmentation using newly generated alleles of the Psen1 mutation. In Psen1-deficient mice, proteolytic activation of Notch1 was significantly affected and the expression of several genes involved in the Notch signalling pathway was altered, including Delta-like3, Hes5, lunatic fringe (Lfng) and Mesp2. Thus, Ps1-dependent activation of the Notch pathway is essential for caudal half somite development. We observed defects in Notch signalling in both the caudal and rostral region of the presomitic mesoderm. In the caudal presomitic mesoderm, Ps1 was involved in maintaining the amplitude of cyclic activation of the Notch pathway, as represented by significant reduction of Lfng expression in Psen1-deficient mice. In the rostral presomitic mesoderm, rapid downregulation of the Mesp2 expression in the presumptive caudal half somite depends on Ps1 and is a prerequisite for caudal somite half specification. Chimaera analysis between Psen1-deficient and wild-type cells revealed that condensation of the wild-type cells in the caudal half somite was concordant with the formation of segment boundaries, while mutant and wild-type cells intermingled in the presomitic mesoderm. This implies that periodic activation of the Notch pathway in the presomitic mesoderm is still latent to segregate the presumptive rostral and caudal somite. A transient episode of Mesp2 expression might be needed for Notch activation by Ps1 to confer rostral or caudal properties. In summary, we propose that Ps1 is involved in the functional manifestation of the segmentation clock in the presomitic mesoderm.

Alleles↗

PubMed2001

Development of astrocytes in the mouse embryonic cerebrum tracked by tenascin-C gene expression.

The development of astrocytes in the rodent cerebrum has been considered to occur mainly in the perinatal stage after the end of neurogenesis, and the mode of generation and migration of the astroglial precursors in the embryonic cerebrum remains poorly understood. Tenascin-C (TN-C) is an astroglia-derived extracellular matrix molecule, and its expression can be traced to the ventricular germinative zone of the embryonic cerebrum. TN-C expression may therefore be used as a marker of astroglial precursors prior to the expression of the glial fibrillary acidic protein (GFAP), a marker for the late stage of astroglial differentiation. The development of astrocytes in the mouse embryonic cerebrum was examined by in situ hybridization histochemistry for TN-C mRNA as an early marker. Observations showed that TN-C-positive cells were aligned in the ventricular germinative zone and began to migrate radially at E (embryonic day) 15. They were dispersed into the cortex as early as E17, and some were still in the DNA synthetic phase (S phase) even after lodgement in the cortex. TN-C-positive cells differentiated into GFAP-immunopositive astrocytes in the cortical and subcortical areas at the neonatal stage. The distribution of their active generation sites in the ventricular germinative zone was not homogeneous but displayed a restricted pattern. The mode of their migration towards the cortical and subcortical areas also showed a characteristic pattern that correlated to the radial glial arrangement. These findings indicate that TN-C-positive cells represent a new subset of astroglial precursors which arise directly from the ventricular germinative zone, and that astrogliogenesis from this precursor occurs as early as the period of neurogenesis at the embryonic stage.

Animals↗

PubMed2001

Development of astrocytes in the mouse hippocampus as tracked by tenascin-C gene expression.

Tenascin-C (TN-C) is an astroglia-derived extracellular matrix protein that has been shown to be an early marker for astroglial precursors in the embryonic mouse brain. This study examined astroglial generation, migration, and differentiation in the developing mouse hippocampus by in situ hybridization histochemistry for TN-C mRNA. Special reference was given to the difference in the mode of astroglial development between the two cortical structures of the hippocampus: the dentate gyrus and Ammon's horn. TN-C-positive cells were found in the ventricular germinative zone of the hippocampus as early as the 15th gestational day, and the labeled cells in the zone apposed to the fimbria migrated tangentially through the subpial area towards the forming dentate gyrus. The TN-C-positive cells aligned in the dentate gyrus exhibited the characteristic morphology of unipolar astrocytes as revealed by double labeling with glial fibrillary acidic protein (GFAP)-immunohistochemistry. On the other hand, the TN-C-positive cells ranging over a wide area of the ventricular germinative zone facing the forming Ammon's horn migrated radially towards the cortex, with most of them aligned in the Ammon's horn exhibiting a GFAP-positive stellate morphology. The onset of migration towards the dentate gyrus was two days earlier than that towards the Ammon's horn. TN-C-positive cells in both cortical structures exhibited a DNA-replicating activity after settlement in the early postnatal stage and were considered to further generate astrocytes. On the other hand, TN-C-positive cells with DNA-replicating activity were also found in the subpial migratory stream moving towards the dentate gyrus and were considered to form the subpial matrix for the generation of the dentate astrocytes. Migratory TN-C-positive cells directed towards both the dentate gyrus and Ammon's horn were apposed to radial glial processes and were believed to be guided by contact with these processes in a manner similar to migratory immature neurons. These findings indicate that TN-C-positive cells for the dentate gyrus and those for the Ammon's horn have different migratory patterns and undergo different morphological differentiations depending on their site of origin at the early stage of astrogliogenesis and corresponding to the different modes of neurogenesis in the two cortical structures.

Animals↗

PubMed2000

Interaction of the Unc-51-like kinase and microtubule-associated protein light chain 3 related proteins in the brain: possible role of vesicular transport in axonal elongation.

We identified two mammalian ULK1 (Unc-51-like kinase involved in neurite extension) binding proteins by yeast two-hybrid screening. Both proteins showed high structural similarity to microtubule-associated protein (MAP) light chain 3 (LC3). One is identical to the Golgi-associated ATPase Enhancer of 16 kDa (GATE-16), an essential factor for intra-Golgi transport [39]. The other is identical to the gamma 2-subunit of GABA-A receptor associated protein (GABARAP) which has a possible role in receptor transport [46]. Using the yeast two-hybrid system and the in vitro GST pull-down assay, we found that the N-terminal proline/serine rich (PS) domain of ULK1 (amino acid 287-416) is required for ULK1-GATE-16 and ULK1-GABARAP protein interactions. However, the kinase activity of ULK1 affected neither ULK1-GATE-16 nor ULK1-GABARAP interaction. Immunohistochemical analysis using ULK1 and GABARAP antibodies showed that the ULK1 and the GABARAP proteins co-localized to many kind of neurons such as pyramidal cells of the hippocampus, mitral cells of the olfactory bulb, and Purkinje cells of the cerebellum. In HeLa cells, endogenous ULK1 and tagged GABARAP showed punctate structures in the cytosol, and were colocalized. These results suggest that the interaction of ULK1 and GABARAP is important to vesicle transport and axonal elongation in mammalian neurons.

Adaptor Proteins, Signal Transducing↗

PubMed2000

A new approach for glomerular lesions: evaluation of scanning acoustic microscopy (SAM) for experimental glomerular disease in rats.

Most pathological evaluations using ordinary optical microscopy are usually qualitative and subjective. The beneficial properties of scanning acoustic microscopy (SAM) include not only observation of microstructure but also quantitative measurement of acoustic propagation speed, indicating the tissue elasticity. In this study, we examined the capability of SAM to evaluate pathological findings in glomeruli using anti-Thy.1 glomerulonephritis. Light microscopic observations of the glomeruli showed severe cell proliferation and mesangial matrix expansion at 10 days after induction of glomerulonephritis and, yet, to a lower extent at day 21 with onset of healing. C-mode scanning of SAM enabled imaging of glomerular structure compatible to findings of ordinary light microscopy. In addition, glomerular propagation speed in nephritic rats was significantly increased at day 10, and then decreased at day 21. These results indicate that SAM evaluation may be a new, useful technique for quantitative evaluation of proliferative glomerular lesions.

Animals↗

PubMed2000

Identification of a novel HLA-B46 allele, B*4602, in Japanese.

The B*4602 was identified in a healthy Japanese donor. The B46 variant antigen differed serologically from B46. The nucleotide sequence of B*4602 differed from that of B*4601 by a single base (from A to G) at position 293 in exon 2. The substitution changed the amino acid Asp at position 74 to Gly. The gene frequency of B*4602 in the Japanese population is extremely low. The presumed haplotype of B*4602 was HLA-A2-Cw1-B*4602-DRB1*09. This is the first report of polymorphism of the B46 antigen.

Alleles↗

PubMed2000

Decapitated and decaudated spermatozoa in man, and pathogenesis based on the ultrastructure.

The ultrastructure of decapitated tails and decaudated heads of ejaculated spermatozoa from an infertile man were investigated. The decaudated heads had the nucleus and acrosome, but neither the implantation fossa nor the basal plate were observed at the caudal pole of the nucleus. The nuclear membrane at this portion showed numerous nuclear pores. The decapitated tails contained the proximal centriole at the proximal end. In addition, most decapitated tails had segmented columns, outer coarse fibres, axoneme, mitochondrial sheath and fibrous sheath in the normal position. The ultrastructure reflects the high motility of the decapitated tails of the spermatozoa. Drastic abnormalities were found in the developing spermatids in the biopsied testes. In spermatids later than the Golgi phase, the pair of centrioles were floating in the cytoplasm of the spermatid, but the distal centriole elongated the flagellum. Neither the implantation fossa nor the basal plate were observed at the caudal pole of the nucleus. It is concluded that the tail might be separated from the head before or after completion of tail formation in spermatids. The explanation for why the proximal centriole failed to connect to the nucleus is unknown.

Adult↗

PubMed2000

Sensitive detection of myocardial contraction abnormality in chronic hemodialysis patients by ultrasonic tissue characterization with integrated backscatter.

Since acoustic properties of the myocardium are sensitive to the myocardial structure and the contractile conditions of myocyte, the authors evaluated cardiac dysfunction based on the integrated ultrasonic backscatter in 18 hemodialysis (HD) patients (duration: 102 +/- 84 months, mean age: 57.6 +/- 9.7 years) and 11 age-matched normals. The cyclic variation of integrated backscatter (CV-IB) at interventricular septum (IVS) and left ventricular posterior wall (PW) was measured and compared with percent fractional shortening (%FS) and percent wall thickening (%Th). The CV-IB of HD patients was smaller than that of control subjects (IVS: 6.2 +/- 1.1 dB vs 8.2 +/- 1.1 dB, p = 0.0003 and PW: 8.4 +/- 2.2 vs 10.3 +/- 1.3, p= 0.025). No significant difference was observed in %FS and %Th between HD patients and control subjects. In HD, the ratio of velocities of early diastolic inflow (E) to late atrial inflow was decreased (0.7 +/- 0.2 vs 1.1 +/- 0.7, p = 0.049) and deceleration time of E was prolonged significantly (200 +/- 28 msec vs 159 +/- 30 msec, p = 0.0082). In the absence of overt cardiac systolic dysfunction, myocardial damage indicated as a decrease in CV-IB and diastolic dysfunction identified on transmitral velocity waveform were detected, which may reflect from the myocardial fibrosis. As a mechanism, pressure overload, hyperparathyroidism, and anemia were neglected, and the other humoral factors may contribute to the myocardial damage in chronic renal failure.

Aged↗

PubMed2000

Differential transport properties of D-leucine and L-leucine in the archaeon, Halobacterium salinarum.

The transport of D-leucine was compared with that of L-leucine in Halobacterium salinarum. When a high-outside/low-inside Na+ gradient was imposed, D-leucine as well as L-leucine accumulated in envelope vesicles, supporting the hypothesis that D-leucine is transported via a symport system along with Na+. Kinetic analyses, including inhibition experiments, indicated that both enantiomers are transported via a common carrier. However, a Hill plot indicated a single binding site for Na+ during L-leucine transport, but dual binding sites for Na+ during D-leucine transport. Furthermore, D-leucine transport was dependent on electrical membrane potential, suggesting that a transporter bound with D-leucine is positively charged. L-leucine transport was slightly, if at all, dependent on membrane potential, suggesting that a transporter bound with L-leucine is electrically neutral. These results indicate that the leucine carrier in Halobacterium salinarum translocates two moles of Na+ per mole of D-leucine, and one mole of Na+ per mole of L-leucine.

Biological Transport↗

PubMed2000

[Interference by D-mannitol on serum D-arabinitol determination by enzymatic assay].

We examined a disparity in D-arabinitol values between two commercial assay kits, LABOFIT and ARABINITEC-AUTO. The determined values by the former were increased by 26%(y = 0.2643x) because of concomitant D-mannitol, whereas those by our newly developed ARABINITEC-AUTO was increased only by 2%(y = 0.0242x). Of 109 samples, 5 samples were found to contain more than 100 mumol/l of D-mannitol. A clear relation(r = 0.89) was noted between the degree of disparity between measurements by the two methods and D-mannitol concentrations in samples. Thus, we have proved that the disparity is mainly caused by D-mannitol.

Biomarkers↗

PubMed2000

[A case of aneurysm of the ductus arteriosus in the adult].

Aneurysm of the ductus arteriosus in the adults is rare. A 60-year-old male with no symptoms was admitted to our hospital. 3 D-CT scans and aortograms indicated a diagnosis of aneurysm of the ductus arteriosus. Operation was done through median sternotomy with the aid of partial cardiopulmonary bypass. The saccular aneurysm was located between the aortic isthmus and the left pulmonary artery. The aneurysm was resected and closed a patch. The post-operative course was uneventful.

Aneurysm↗

PubMed1999

Cellular and subcellular localization of a newly identified member of the protein 4.1 family, brain 4.1, in the cerebellum of adult and postnatally developing rats.

For obtaining a deeper insight into the properties of a newly characterized member of the protein 4.1 family, brain 4.1, the cellular and subcellular localization was investigated in the cerebellar cortex of adult and postnatally developing rats. Fluorescent immunohistochemical observations showed that brain 4.1 localized predominantly to glomeruli in the granular layer and throughout the molecular layer in adult rat cerebellar cortex. Analysis of subcellular localization of brain 4.1 by immuno-electron microscopy further demonstrated that presynaptic terminals of mossy fibers and parallel fibers, cytoplasm of granule cells and cytoplasm and/or processes of glial cells contained brain 4.1 while postsynaptic regions of the dendrites of granule cells and Purkinje cells, axons and myelin sheaths did not. Thus, one of the major subcellular destination of brain 4.1 was presynaptic terminal in the cerebellum. This was further supported by the fact that the immunostaining pattern of brain 4.1 in the cerebellum changed in a similar way to that of a synaptic terminal marker, synaptophysin during the postnatal development. Immunoblot analysis also demonstrated that contents of brain 4.1 isoforms varied in parallel with the changes of the immunostaining pattern. Biochemical analysis confirmed the presence of brain 4.1 at synaptic terminals, but there was no obvious correlation between each isoform and its subcellular localization. These results suggested that brain 4.1 is involved in the formation and maintenance of synapse as a membrane skeletal component at presynaptic terminals in the cerebellum.

Aging↗

PubMed1999

Notochord-dependent expression of MFH1 and PAX1 cooperates to maintain the proliferation of sclerotome cells during the vertebral column development.

During axial skeleton development, the notochord is essential for the induction of the sclerotome and for the subsequent differentiation of cartilage forming the vertebral bodies and intervertebral discs. These functions are mainly mediated by the diffusible signaling molecule Sonic hedgehog. The products of the paired-box-containing Pax1 and the mesenchyme forkhead-1 (Mfh1) genes are expressed in the developing sclerotome and are essential for the normal development of the vertebral column. Here, we demonstrate that Mfh1 like Pax1 expression is dependent on Sonic hedgehog signals from the notochord, and Mfh1 and Pax1 act synergistically to generate the vertebral column. In Mfh1/Pax1 double mutants, dorsomedial structures of the vertebrae are missing, resulting in extreme spina bifida accompanied by subcutaneous myelomeningocoele, and the vertebral bodies and intervertebral discs are missing. The morphological defects in Mfh1/Pax1 double mutants strongly correlate with the reduction of the mitotic rate of sclerotome cells. Thus, both the Mfh1 and the Pax1 gene products cooperate to mediate Sonic hedgehog-dependent proliferation of sclerotome cells.

Animals↗

PubMed1999

A novel type of non-coding RNA expressed in the rat brain.

We have characterized a novel type of non-coding RNA which consists of tandem repeats of similar sequences, approximately 0.9 kb in size. This RNA, termed Bsr (brain specific repetitive) RNA, is encoded at a single locus (6 q31-->q32) in the rat genome, where 100 to 150 copies of the 0.9 kb sequences are repeated in tandem. Bsr RNA is preferentially expressed in the rat central nervous system (CNS), especially in phylogenetically old structures, such as the pareo- and archicortex, amygdala, thalamus and hypothalamus. In the developing brains, Bsr RNA is expressed in the subsets of differentiating cells but not in proliferating cells. Despite the finding that Bsr RNA appears to be conserved only among the Rattus species, the specific expression pattern of Bsr RNA suggests that it might have some role in the rat CNS.

Animals↗

PubMed1999

Immunohistochemical localization of a water channel, aquaporin 7 (AQP7), in the rat testis.

Cell volume reduction is one of the most distinct morphological changes during spermiogenesis and may be largely attributable to water efflux from the cell. A strong candidate for a water efflux route, aquaporin 7 (AQP7), which is a water channel, was studied immunohistochemically in the rat testis. Immunoreactivity was restricted within the elongated spermatids, testicular spermatozoa, and residual bodies remaining in the seminiferous epithelium. Weak but distinct immunoreactivity was first observed in the cytoplasmic mass of the spermatid at step 8 of spermiogenesis. The Golgi-like apparatus became steadily immunoreactive at step 10. The plasma membrane covering the cytoplasmic mass showed strong immunoreactivity after step 16. At this step, the middle piece of the tail also showed immunoreactivity at the portion protruding into the lumen. The whole head and distal tail, where the elongated spermatid had only a limited amount of cytoplasm, showed no immunoreactivity throughout spermiogenesis. After spermiation, the immunoreactivity of AQP7 remained at the middle piece and in the cytoplasmic droplet in the testicular spermatozoon. The present observations suggest that AQP7 contributes to the volume reduction of spermatids, since this water channel protein is localized on the plasma membrane covering the condensing cytoplasmic mass of the elongated spermatid, and since the seminiferous tubule fluid is hypertonic.

Animals↗

PubMed1999

Histological characterization of defective spermatogenesis in mice lacking the basigin gene.

Basigin is a transmembrane protein belonging to the immunoglobulin superfamily. In the light of the fact that knockout mice lacking the basigin gene (Bsg) are azoospermic, the phenotype in the male reproductive system was extensively examined in this study. Spermatogenesis in Bsg (-/-) mice was found to be disrupted, and arrested at the metaphase of the first meiotic division. A few germ cells differentiated into young spermatids, but they were exfoliated. The lumens of the male reproductive system were filled with round degenerated cells. Using the TUNEL method and electron microscopy, some of the degenerated cells in the testis and epididymal head were shown to be apoptotic. Crystalloids of fine tubules and unusual ectoplasmic specializations were also observed in the Sertoli cells of Bsg (-/-) mice. These specializations displayed unusual 'circular' structures. Furthermore, unusual ectoplasmic specializations covering the spermatocytes rather than the mature spermatids were found. These structures were formed as a result of the lack of mature spermatids in the Bsg (-/-) testis. Results from analyses of azoospermia in the Bsg (-/-) mice suggest that basigin, through the interactions between germ cells and Sertoli cells, is an essential factor in the growth and/or survival of spermatids.

Animals↗

PubMed1999

The effect of combination therapy with interferon and cryofiltration on mesangial proliferative glomerulonephritis originating from mixed cryoglobulinemia in chronic hepatitis C virus infection.

Cryofiltration, which has developed from double filtration plasmapheresis (DFPP) with a cooling unit, is an on-line technique to remove cryoglobulin. We report on a patient who suffered from progressive edema and renal insufficiency caused by cryoglobulinemic membranoproliferative glomerulonephritis (MPGN), probably due to chronic hepatitis C virus (HCV) infection. To remove cryoglobulins and terminate the HCV infection, we utilized combination therapy with cryofiltration and interferon-alpha injection with corticosteroids. Interferon-alpha was capable of decreasing proteinuria but not diminishing cryoglobulin. Additional cryofiltration could remove cryoglobulin to an undetectable level. This combination therapy was partially successful to reduce proteinuria and prevent the progressive deterioration of renal function. The major adverse effects of this therapy were bleeding and myelosuppression. We conclude that this combination therapy may be effective and should be considered as treatment for cryoglobulinemic MPGN.

Aged↗