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Yoriko Atomi

Publications and source records attributed to Yoriko Atomi.

10 recordsLinked to original sources
PubMed2005

The decrease of the cytoskeleton tubulin follows the decrease of the associating molecular chaperone alphaB-crystallin in unloaded soleus muscle atrophy without stretch.

The cytoskeletal component tubulin/microtubule commonly allows the cell to respond mechanically to the environment. The concentration of free tubulin dimer is autoregulated in the balance of free dimer and polymeric forms of microtubule (MT) protein, having an intrinsic property of "dynamic instability", and through cotranslational beta-tubulin mRNA degradation. Recently, we have demonstrated that alphaB-crystallin is a key molecule of muscle atrophy, since alphaB-crystallin has a chaperone-like-activity that suppresses tubulin aggregation and protects the MT disassembly against both Ca2+ and depolymelizing alkaloid in vitro. Most of the small heat-shock proteins (sHsps), including alphaB-crystallin, are expressed in skeletal muscle. However, no report to date has studied the changes of tubulin/MT during muscle adaptation. Here, we examined changes in tubulin content in rat soleus muscles after hindlimb suspension (HS) with/without passive stretch and the recovery. HS induced rapid decreases of soleus muscle mass, most Hsps (alphaB-crystallin, Hsp90, Hsp70, Hsp27, and p20) and tubulin contents in soleus muscle, while heat-shock cognate 70-kDa protein (Hsc70) did not decrease. Soleus muscle mass, most Hsps, and tubulin were maintained with passive stretch. After 5 days' recovery, the levels of tubulin and Hsps, but not Hsc70, were restored to control levels. The interactions of alphaB-crystallin and tubulin/MT were observed with immunoprecipitation with an anti-alpha-tubulin antibody and taxol-dependent MT assembly. Other sHsps were also associated with alphaB-crystallin and MT, whereas Hsp90 and Hsp70 did not co-precipitate with them. These data imply an interaction and close relationship between alphaB-crystallin and tubulin/MTs in muscle tissues. The amount of mRNA of alphaB-crystallin decreased with the muscle atrophy level, whereas the gene expression level of betaI-tubulin was maintained during HS. This means a significant role of post-transcriptional regulation in tubulin/MT system in muscle adaptation, whereas alphaB-crystallin and most sHsps are regulated at the transcriptional level. Additional functional contribution of alphaB-crystallin to tubulin/MTs during myotube formation was examined using C2C12 myoblast cultured cells, the alphaB-crystallin expression of which was decreased or increased. It indicated the necessity of alphaB-crystallin during microtubule reorganization. In conclusion, tubulin/MTs were revealed to be one of the substrates of alphaB-crystallin, and also serial decreases of alphaB-crystallin and tubulin/MT in early soleus muscle atrophy suggest that the chaperone effect of alphaB-crystallin on the cytoskeleton, which may be also dynamically regulated in the muscle cell, is a key mechanism for muscle adaptation and protection of the atrophy and also muscle differentiation.

Animals↗
PubMed2004

alphaB-Crystallin-coated MAP microtubule resists nocodazole and calcium-induced disassembly.

alphaB-Crystallin, one of the small heat-shock proteins, is constitutively expressed in various tissues including the lens of the eye. It has been suggested that alphaB-crystallin provides lens transparency but its function in nonlenticular tissues is unknown. It has been reported that alphaB-crystallin is involved in the stabilization and the regulation of cytoskeleton, such as intermediate filaments and actin. In this study, we investigate the possibility whether alphaB-crystallin interacts with the third cytoskeleton component, microtubules (MTs). First, we precisely observed the cellular localization of alphaB-crystallin and MT networks in L6E9 myoblast cells and found a striking coincidence between them. MTs reconstituted from cell lysate contained alphaB-crystallin. Electron micrographs clearly showed direct interactions of purified alphaB-crystallin with the surface of microtubule-associated proteins (MAPs) attached to MTs. Purified alphaB-crystallin bound to MAP-MTs in a concentration-dependent manner. However, alphaB-crystallin did not bind MTs reconstituted from purified tubulin. Finally, we observed that alphaB-crystallin increased the resistance of MTs to depolymerization in cells and in vitro. Taken together, these results suggest that one of the functions of alphaB-crystallin is to bind MTs via MAP(s) and to give the MTs resistance to disassembly.

Animals↗
PubMed2004

The change of HSP47, collagen specific molecular chaperone, expression in rat skeletal muscle may regulate collagen production with gravitational conditions.

It is well known that unloading of skeletal muscle with spaceflight leads skeletal muscle atrophy. However, it remains unclear how the extracellular matrix within the muscle and the connective tissues such as tendon and ligament respond to reduced mechanical load including microgravity, although they have been thought to play important roles in both the transmission of force and the signal transduction between cells and tissues. Type-I collagen and type-IV collagen, both of the major components of extracellular matrix and connective tissues. We focused on change of these collagen synthesis with mechanical load. To obtain an insight into the effects of gravitational changing on the protein metabolism of collagen in skeletal muscle during mechanical unloading, reloading after unloading, we investigated changes in the amount of Heat shock protein 47 (HSP47), has been postulated to be a collagen-specific molecular chaperone localized in the ER (Nagata et al, 1992). Western blot analysis revealed that HSP47 in rat soleus muscle decreases at 5 days after hindlimb suspension (HS). On the other hand, HSP47 in rat soleus muscle increases at 5 days after hypergravity (HG) induced by the centrifugation. RT-PCR analysis showed HSP47 mRNA decreased with HS earlier, as compared with collagen type-I and type-IV mRNA. From these results, the amount of HSP47 changing by gravitational condition may effect on signal transfers in the primary stage of adaptation and the change of HSP47 expression in skeletal muscle may regulate collagen production with gravitational conditions.

Animals↗
PubMed2003

Physical performance and soleus muscle fiber composition in wild-derived and laboratory inbred mouse strains.

We compared four inbred mouse strains in their physical performance, measured as a maximal treadmill running time, characteristics of soleus muscle, anatomic character, and growth. The strains used were Mus musculus domesticus [C57BL/6 (B6) and BALB/c], Mus musculus molossinus (MSM/Ms), and Mus spretus. Maximal running time was significantly different among these four mouse strains. Running time until exhaustion was highest in MSM/Ms and lowest in M. spretus. Maximal times for the laboratory mouse strains were nearly identical. Soleus muscle fiber type and cross-sectional area also differed significantly among the species. In particular, M. spretus was significantly different from the other inbred mouse strains. Growth in the wild-derived inbred mice appeared to be complete earlier than in the laboratory mice, and the body size of the wild strains was about half that of the laboratory strains. From these results, we propose that wild-derived inbred mouse strains are useful models for enhancing phenotypic variation in physical performance and adaptability.

Anatomy, Cross-Sectional↗
PubMed2003

Suppression of cofilin phosphorylation in insulin-stimulated ruffling membrane formation in KB cells.

Various cellular events such as cell motility and division are directed by the actin cytoskeleton under the control of its regulatory system. Cofilin is a low molecular weight actin-modulating protein that severs and depolymerizes F-actin and is shown to enhance actin filament dynamics. The activity of cofilin is negatively regulated by phosphorylation at Ser-3. In human epidermoid carcinoma KB cells, insulin treatment induces characteristic ruffling membranes, and it was reported that LIMK1, a cofilin kinase, was activated in these cells treated with insulin. Since cofilin is a key protein responsible for establishing the rapid turnover of actin filaments, it appears to be contradictory that cofilin is phosphorylated (inactivated) by a stimulus that is known to induce the highly dynamic actin structure, ruffling membranes. Therefore, we examined the phosphorylation state of endogenous cofilin in KB cells treated with insulin. The dephosphorylated form of cofilin increased with insulin treatment, as analyzed by nonequilibrium pH gradient gel electrophoresis (NEpHGE)-immunoblotting. Cell labeling with (32)P orthophosphate indicated that cofilin was being continuously phosphorylated and dephosphorylated, and that the apparent insulin-induced dephosphorylation was due to suppression of continuous phosphorylation and not to enhanced dephosphorylation. Further, we examined the localization of the phosphorylated form of cofilin using phospho-specific antibody raised against phosphorylated cofilin. Surprisingly, phosphorylated cofilin was concentrated in the ruffling membranes induced by insulin. These results suggest that the examination of the kinetics and spatial regulation of phosphorylation is critical for the elucidation of the role of cofilin and upstream kinases in actin reorganization.

Actin Cytoskeleton↗
PubMed2003

The content of heat shock protein 47 (HSP47), a collagen-specific stress protein, changes with gravitational conditions in skeletal muscle.

It is well known that unloading of skeletal muscle with spaceflight or tail suspension leads rat soleus muscle atrophy. Previously, we reported that one of small heat shock protein (sHSP), alpha B-crystallin shows an early dramatic decrease in atrophied rat soleus muscle (Atomi et al, 1991). In this report, we focused to study the gravitational responses of another HSP, which may be reactive to the gravity. HSP47, a collagen-specific stress protein, has been postulated to be a collagen-specific molecular chaperone localized in the ER (Nagata et al, 1992). Western blot analysis revealed that HSP47 in slow skeletal muscle decreases at 5 days after tail suspension (TS) and increased at 5 days recovery after 10 days of TS as compared with the control level. Hypothetically, HSP47 in slow soleus muscle increases at 5 days after hypergravity (HG) induced by the centrifugation. The content of HSP47 in soleus muscle was strongly affected by gravity conditions.

Animals↗
PubMed2002

Gene expression of type I and type III collagen by mechanical stretch in anterior cruciate ligament cells.

Mechanical stretch affects the healing and remodeling process of the anterior cruciate ligament (ACL) after surgery in important ways. In this study, the effects of mechanical stress on gene expression of type I and III collagen by cultured human ACL cells and roles of transforming growth factor (TGF)-beta1 in the regulation of mechanical strain-induced gene expression were investigated. Uniaxial cyclic stretch was applied on ACL cells at 10 cycles/min with 10% length stretch for 24 h. mRNA expression of the type I and type III collagen was increased by the cyclic stretch. TGF-beta1 protein in the cell culture supernatant was also increased by the stretch. In the presence of anti-TGF-beta1 antibody, stretch-induced increase in type I and type III mRNA expression was markedly ablated. The results suggest that the stretch-induced mRNA expression of the type I and type III collagen is mediated via an autocrine mechanism of TGF-beta1 released from ligament cells.

Anterior Cruciate Ligament↗
PubMedDate not supplied

Proliferation and differentiation of Xenopus A6 cells under hypergravity as revealed by time-lapse imaging.

Xenopus laevis A6 cells, which are cloned epithelial cells from the Xenopus kidney, differentiate into a dome structure when the cells reach confluence. We investigated the gravitational responses of A6 cellular motility during normal differentiation and differentiation under hypergravity conditions using centrifugation (1-100 x g). Progression to dome formation was analyzed by time-lapse micrography. Dome formation and increased expression of Na(+)/K(+)-adenosine triphosphatase were used as markers of differentiation. Interestingly, a high rate of cellular proliferation was observed at a low level of hypergravity (5 x g). Despite this, there was no difference in the time to dome formation between the control cells at primary cell density and those that differentiated under hyper- or hypogravity conditions. In conclusion, this experiment on amphibian cells revealed that the proliferation of A6 cells was strongly affected by gravity conditions, but the differentiation step appears to be controlled by an intra- or intercellular clock.

Animals↗