[Differentiation of muscle proteins and their significance].
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Actins↗
Biomedical subjects
T Masaki
Publications and source records attributed to T Masaki.
[Abnormal cell membrane and muscle cell differentiation disorder in muscular dystrophy].
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Animals↗
Immunochemical analysis of C-protein isoform transitions during the development of chicken skeletal muscle.
Isoforms of C-protein in adult chickens which differ in fast (pectoralis major, PM) and slow (anterior latissimus dorsi, ALD) skeletal muscles can be distinguished immunochemically with monoclonal antibodies (McAbs) specific for the respective fast (MF-1) and slow (ALD-66) protein variants (Reinach et al., 1982 and 1983). The expression of these C-proteins during chick muscle development in vivo has been analyzed by immunoblot and immunofluorescence procedures. Neither MF-1 nor ALD-66 reacted with whole-cell lysates or myofibrils from PM of 12-day-old embryos. However, both McAbs bound to peptides of 145 kDa in PM from late embryonic and young posthatched chickens. All of the myofibers in these muscles reacted with both antibodies, but the binding of the anti-slow McAb (ALD-66) diminished progressively with age and was completely negative with PM by 2 weeks after hatching. In contrast, the ALD muscle from 17 days in ovo thru adulthood only reacted with ALD-66; no binding of MF-1 could be detected at these stages. Since both fast and slow myosin light chains (LC) coexist within embryonic pectoralis and ALD muscles (e.g., G. F. Gauthier, S. Lowey, P. A. Benfield, and A. W. Hobbs, 1982, J. Cell Biol. 92, 471-484) yet segregate to specific fast and slow muscle fibers at different stages of development, the temporal transitions of C-protein and myosin LC were compared during myogenesis. "Slow-type" C-protein appeared after the disappearance of slow myosin light chains, whereas the accumulation of the "fast-type" light chains occurred before the expression of "fast-type" C-protein. The pattern of isoform transitions appears to be far more complex than previously suspected.
Age Factors↗
Coexistence of fast-type and slow-type C-proteins in neonatal chicken breast muscle.
Two different C-protein variants which selectively react with either monoclonal anti-fast C-protein antibody (MF-1) or monoclonal anti-slow C-protein antibody (ALD-66) were separated from neonatal chicken pectoralis muscle by hydroxylapatite column chromatography. Myofibrils isolated from the neonatal chicken muscle reacted with both monoclonal antibodies as examined by an indirect immunofluorescence method. These observations strongly indicate that both fast-type and slow-type C-proteins are expressed in the neonatal chicken skeletal muscle. Both of them are intermingled and assembled in the same myofibrils.
Aging↗
Differential expression and distribution of chicken skeletal- and smooth-muscle-type alpha-actinins during myogenesis in culture.
Antibodies to chicken fast skeletal muscle (pectoralis) alpha-actinin and to smooth muscle (gizzard) alpha-actinin were absorbed with opposite antigens by affinity chromatography, and four antibody fractions were thus obtained: common antibodies reactive with both pectoralis and gizzard alpha-actinins ([C]anti-P alpha-An and [C]anti-G alpha-An), antibody specifically reactive with pectoralis alpha-actinin ([S]anti-P alpha-An), and antibody specifically reactive with gizzard alpha-actinin ([S]anti-G alpha-An). In indirect immunofluorescence microscopy, (C)anti-P alpha-An, (S)anti-P alpha-An, and (C)anti-G alpha-An stained Z bands of skeletal muscle myofibrils, whereas (S)anti-G alpha-An did not. Although (S)anti-G alpha-An and two common antibodies stained smooth muscle cells, (S)anti-P alpha-An did not. We used (S)anti-P alpha-An and (S)anti-G alpha-An for immunofluorescence microscopy to investigate the expression and distribution of skeletal- and smooth-muscle-type alpha-actinins during myogenesis of cultured skeletal muscle cells. Skeletal-muscle-type alpha-actinin was found to be absent from myogenic cells before fusion but present in them after fusion, restricted to Z bodies or Z bands. Smooth-muscle-type alpha-actinin was present diffusely in the cytoplasm and on membrane-associated structures of mononucleated and fused myoblasts, and then confined to membrane-associated structures of myotubes. Immunoblotting and peptide mapping by limited proteolysis support the above results that skeletal-muscle-type alpha-actinin appears at the onset of fusion and that smooth-muscle-type alpha-actinin persists throughout the myogenesis. These results indicate (a) that the timing of expression of skeletal-muscle-type alpha-actinin is under regulation coordination with other major skeletal muscle proteins; (b) that, with respect to expression and distribution, skeletal-muscle-type alpha-actinin is closely related to alpha-actin, whereas smooth-muscle-type alpha-actinin is to gamma- and beta-actins; and (c) that skeletal- and smooth-muscle-type alpha-actinins have complementary distribution and do not co-exist in situ.
Actinin↗
Proximity of reactive lysyl residue to the antigenic site in rabbit skeletal myosin against the monoclonal antibody (MF-18) generated to chicken skeletal myosin.
MF-18, one of the monoclonal antibodies generated to chicken myosin, cross-reacted with rabbit skeletal myosin subfragment-1 (S1). Utilizing an improved procedure of immuno-blotting, a decrease in reactivity of MF-18 to S1 by trinitrophenylation was observed. This indicates that the reactive lysyl residue is very close to the hapten site. This is consistent with the evidence that the hapten site resides in the 26,000 dalton tryptic fragment of S1. Use of such antibodies as labels may open the way to determining the location of specific hapten sites in the three-dimensional image of actin-S1 complex reconstructed from the electron micrographs.
Animals↗
Biphasic release of thyrotropin in response to thyrotropin-releasing hormone (TRH) from rat anterior pituitary cells in vitro: possible dependence on protein synthesis.
Mechanisms related to the biphasic release of TSH were studied using primary cultured cells of the rat anterior pituitary gland on micro-carrier beads in a superfusion system. Release of TSH in response to continuous exposure to TRH exhibited a biphasic pattern; the first phase was characterized by a rapid, transient and high-rate release (phase I) and the second phase by a chronic and low-rate release (phase II). The shift of the release from phase I to phase II occurred by treatment with TRH at concentrations from submaximal to maximal. When the Ca2+ concentration in the medium was decreased, the phase I release was partially inhibited, while the phase II release was completely inhibited, suggesting a difference between the mechanisms in phase I and phase II release. The phase I release was not suppressed by cycloheximide. This protein synthesis-independent release of phase I seemed to be linked to the intracellular releasable pool of TSH. The phase II release was suppressed by the presence of a protein synthesis inhibitor. After the phase II release was suppressed by cycloheximide, the magnitude of phase I release in response to re-exposure to TRH markedly decreased. The decreased phase I release in response to TRH was observed with the cells which were previously stimulated by high K+ instead of TRH, suggesting that the decrease in the response of phase I reflects the depletion of a releasable pool of TSH rather than homologous desensitization of thyrotrophs with TRH. These results suggest that the phase I release of TSH depends on a portion of the previously prepared-releasable pool while phase II release depends on previously prepared plus newly prepared pools of TSH. Replenishment of the releasable TSH pool was considered to involve protein synthesis.
Animals↗
Rabbit hearts for the critical evaluation of drugs to reduce the size of experimentally produced acute myocardial infarction.
Using more than 500 rabbits, we found that the rabbit heart is a good model for the evaluation of drugs which affect acute myocardial infarction (AMI) size. When the ratio of the epicardial coloration area to the long axis length of the left ventricle was controlled immediately after the ligation of the left anterior descending coronary artery and small branches of the left circumflex artery, it was possible to estimate the size of the ischemic region because AMI region in rabbit heart was always transmural. The necrotic region in the left ventricle was determined by phosphorylase histochemistry 24 hours after the operation. The incidence of arrhythmia and death following the operation was negligible. Then, we evaluated several drugs to examine their effects on AMI size. Propranolol (1, 2, and 4 mg/Kg) and verapamil (0.25, 0.5, and 1.0 mg/Kg) reduced AMI size, although the mortality and AMI size increased at higher doses of verapamil. Another Ca2+ antagonist, diltiazem (2 mg/Kg) and an adenosine potentiator, dilazep (2 mg/Kg) also decreased AMI size, while nicardipine, a water soluble, photoresistant nifedipine analogue (0.01, 0.05, and 0.1 mg/Kg) did not show a significant effect. These data suggest that this rabbit model is useful for assessing drug effects on AMI size and that the mechanism(s) of action of nicardipine may differ from other Ca2+ antagonists.
Animals↗
[Combination chemotherapy with neocarzinostatin(NCS), HCFU and picibanil(NHO therapy) for advanced carcinoma of the digestive system--a comparative study with NF, and NFO therapy].
We have previously reported the clinical effects of NF therapy (NCS + 5-FU) and NFO therapy (NCS + 5-FU + Picibanil) on patients with advanced carcinoma of the digestive organs. In the present study, (NHO therapy (NCS + HCFU + Picibanil) performed in 41 patients and 30 patients were evaluated for its clinical effects. In comparison with NHO, NF and NFO, partial regression (tumor regression exceeding 50%) was noted in 5 of 30 patients (16.7%) on NHO, which was superior to 7.4% on NF, but slightly inferior to 18.8% on NFO. However, six and twelve month survival rate and 50% survival month on NHO therapy were 31.6%, 10.5% and 4.6 months, respectively and they were superior to those of NF and NFO therapy. Though the incidence of the adverse effects by NHO was almost identical with that of NFO and not more frequent than that of NF therapy. Urinary frequency, hot sensation and urgency due to HCFU administration were observed approximately in 10% on NFO therapy. In the three modalities the advantageous clinical effects on patients with hepatic carcinoma irrespective of primary or metastatic were observed.
Adult↗
Effect of parathyroid hormone, cyclic AMP and Ca2+ on the phosphorylation of brush border membranes in rabbit kidney.
Renal cortical slices were incubated with parathyroid hormone or dibutyryl cAMP and the effects on phosphate uptake and phosphorylation of proteins in brush border membranes isolated from the treated slices were determined. Na+ gradient-dependent phosphate uptake was inhibited. Phosphorylation of proteins of Mr=170 K, 135 K, 105 K, 88 K, and 68 K was increased after incubation with the hormone or the cyclic nucleotide. Phosphorylation of membrane proteins was also examined in isolated relatively intact brush border membrane vesicles and in membrane vesicles disrupted with Triton X-100. With intact membrane vesicles, total phosphorylation of the membrane was not significantly altered by cAMP. However, phosphorylation of proteins of Mr=85 K and 48 K increased whereas phosphorylation of proteins of Mr=170 K, 78 K, and 56 K decreased, relative to that found with slices. With detergent-treated membranes, which presumably were made permeable to [gamma-32P]-ATP, a cAMP-induced increase in total phosphorylation was demonstrated. Phosphorylation of proteins of Mr=135 K, 78 K, 65 K, and 56 K was markedly enhanced. These findings suggest that proteins of Mr=135 K, 78 K, 65 K, and 56 K are localized on the cytosolic side of the membrane whereas proteins of Mr=85 K and 48 K are present on the luminal surface of the membrane. Incubation of the isolated brush border membrane vesicles with Ca2+, or Ca2+ plus cAMP, also affected the phosphorylation of membrane proteins. The phosphorylation of proteins of Mr=105 K, 68 K, and 20 K was increased by Ca2+. The Mr=20 K protein may be myosin light chain.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
[An anatomical study of the interphalangeal sesamoid bone of the hallux].
The existence of sesamoid bone on the interphalangeal joint of the hallux has not been sufficiently recognized. The frequency of this sesamoid bone has been described to be low probability in the orthopaedic and anatomical literature. We have, however, experienced some cases of the interphalangeal joint dislocation giving difficulty to usual manipulative reduction because of the presence of the sesamoid bone. In order to ascertain the existence of sesamoid bone on the interphalangeal joint of the hallux, anatomical and radiographic study have been performed with 144 feet of 73 adult cadavers, 32 feet of 16 fetal cadavers and 958 patients. The following results were obtained: The sesamoid bone was seen in 90.9% by radiographical examination and in 95.9% by macroscopic observation for 144 feet on 73 adult cadavers. Cartilagenous sesamoid bone was found in 93.8%, 32 feet of 16 fetal cadavers using stereoscopic microscope. In the radiographical examination of 958 feet of the patients, the sesamoid bones were approved in 56.3%, but it increased to 93.0% using 1/4 sensitivity intensifying screen. The sesamoid bone of the interphalangeal joint on adult cadavers was located at the central portion in parallel with interphalangeal joint space and buried in the fibrocartilagines plantares. This sesamoid bone had two cartilagenous joint surfaces against the base of distal phalanx and the head of proximal phalanx. The sesamoid bone had the shape of longitudinal ellipse and the length, width and thickness were measured. The size of sesamoid bone on the male cadaver were larger than that of the female on the three parameters and there were no special difference between the right and the left sides.
Adolescent↗
[The change in EIB after repeated exercise].
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Adolescent↗
[Anatomical study of the root canal of anterior teeth--with special reference to the marrow canal and apical ramifications].
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Cuspid↗
Characterization of the C-protein from posterior latissimus dorsi muscle of the adult chicken: heterogeneity within a single sarcomere.
Specific isoforms of myofibrillar proteins are expressed in different muscles and in various fiber types within a single muscle. We have isolated and characterized monoclonal antibodies against C-proteins from slow tonic (anterior latissimus dorsi, ALD) and fast twitch (pectoralis major) muscles of the chicken. Although the antibody against "fast" C-protein (MF-1) did not bind to the "slow" isoform and the antibody to the "slow" C-protein (ALD-66) did not bind to the "fast" isoform, we observed that both antibodies bound C-protein from the posterior latissimus dorsi (PLD) muscle. Here we demonstrate that in the PLD muscle the binding sites of these two antibodies reside in two different C-protein isoforms which have different molecular weights and can be separated by hydroxylapatite column chromatography. Since we have shown previously that both these antibodies stain all myofibers and myofibrils derived from PLD muscle, we conclude that all myofibers in this muscle contain both isoforms with all sarcomeres.
Animals↗
Changes in myosin isozymes during development of chicken gizzard muscle.
The distribution of myosin isozymes in embryonic and adult chicken gizzard muscle were examined by electrophoresis in a non-denaturing gel system (pyrophosphate acrylamide gel electrophoresis), and both light and heavy chains of embryonic and adult myosin isozymes were compared. In pyrophosphate acrylamide gel electrophoresis, there were three isozyme components in embryonic gizzard myosin, but only one isozyme in adult gizzard myosin. The mobility of the fastest migrating embryonic isozyme was similar to that of the adult isozyme. The three embryonic isozymes differ from each other in the light chain distribution. Two of them contain an embryo-specific myosin light chain, which is characterized by its molecular weight and isoelectric point, whereas the other embryonic myosin isozyme contained the same light chains as the adult myosin. The pattern of peptide fragments of embryonic heavy chain produced by digestion with alpha-chymotrypsin in the presence of SDS was not distinguishable from that of adult myosin heavy chain. Thus there are myosin isozymes specific to embryonic gizzard muscle which exhibit embryo-specific light chain compositions, but are similar to adult gizzard myosin in their heavy chain structure.
Animals↗
Increase in norepinephrine-induced formation of phosphatidic acid in rat vas deferens after denervation.
Surgical denervation of rat vas deferens causes supersensitivity in that the tissue sensitivity and the maximum response to a variety of agonists increase. To understand the molecular mechanism of supersensitivity in smooth muscle, norepinephrine(NE)-induced alteration in phospholipid metabolism was studied using control and denervated vasa deferentia. When the tissue was stimulated by NE, only [32P]Pi incorporation into phosphatidic acid(PA) was increased in proportion to the increase in NE concentration without any significant effect on that into other phospholipids. This PA labeling was significantly accelerated by denervation. In the denervated tissue, PA labeling was stimulated by lower concentrations of NE and the maximum response to NE was increased compared to the control. The breakdown of phosphatidylinositol 4-monophosphate(DPI) and phosphatidylinositol 4,5-diphosphate (TPI) was also accelerated by NE. But the influence of denervation on this NE-induced DPI and TPI was not marked. Therefore, it is likely that denervation clearly enhanced NE-induced PA labeling without an appreciable effect on that of the other phospholipids. Furthermore, the absolute amount of PA was also increased by NE, and this increase was exaggerated by denervation. Considering that PA can behave as a Ca2+ ionophore in the plasma membrane, these results suggest that the stimulated accumulation of PA plays an important role in receptor-linked supersensitivity in smooth muscle.
Animals↗
Coexistence of cardiac-type and fast skeletal-type myosin light chains in embryonic chicken cardiac muscle.
Myosin from embryonic chicken ventricle contained a light chain component which comigrated with fast skeletal myosin light chain 1 (Lf1) on two dimensional electrophoresis in addition to cardiac type light chains (Lc1 and Lc2). Immunoblot analysis showed that this minor light chain band reacted with anti-Lf1 antibody. Antigens binding with anti-Lc1 and anti-Lf1 antibodies were located on myofibrils in embryonic cardiac muscle cells in vivo and in vitro. From these observations, we conclude that a small amount of Lf1 exists in embryonic chicken cardiac muscle.
Animals↗