Preliminary experiments on mechanical stretch-induced activation of skeletal muscle satellite cells in vivo

We have shown in vitro that mechanical stretch triggers activation of quiescent satellite cells of skeletal muscle to enter the cell cycle through an intracellular cascade of events including nitric oxide (NO) synthesis that results in the release of hepatocyte growth factor (HGF) from its extracellular association and its subsequent presentation to signaling receptors. In order to explore the activation mechanism in vivo, stretch experiments were conducted in the living animal using our suspension model developed. This system used the weight of the hind portion of rats to stretch the inside muscles of the left hind limb suspended for a period of 0.5–2.0 h. At the end of the stretch period, the rats received an intraperitoneal injection of bromodeoxyuridine followed by immunocytochemistry for its incorporation as an index of satellite cell activation in vivo. Depending on the period of stretch, bromodeoxyuridine labeling was increased significantly over the contralateral unstretched leg or control muscle from untreated rats. A stretched muscle extract prepared from the 2 h stretched tissue by incubating it in PBS, showed the active form of HGF as revealed by immunoblotting and it could stimulate the activation of unstretched satellite cells. Also, administering NO synthase inhibitor L‐NAME prior to muscle stretch abolished the stretch activation of satellite cells. Therefore, the results from these experiments demonstrate that stretching muscle triggers NO synthesis and HGF release, which could activate satellite cells in vivo.     

Identification of a telomere sequence type in three sponge species (Porifera) by fluorescence <Emphasis Type="Italic">in situ</Emphasis> hybridization analysis

The telomere sequence type (TTAGGG)_n is known to be distributed in various phyla in the Animalia and in Mastigophora (Protista). However, the telomere type of Porifera (sponges), a phylum comprising the lowest multicellular animals, has not been reported. In this study, we examined the three sponge species Leucetta chagosensis, Halichondria japonica, and Halichondria panicea for the presence of the telomere type (TTAGGG)_n by fluorescence in situ hybridization (FISH). The oligonucleotide probe (TTAGGG)₇ clearly displayed signals on the interphase nuclei of all three sponges. In contrast, the (TTAGG)₇ probe, which has one base fewer than (TTAGGG)₇, did not display the signals. These results suggest that the telomeres of the three sponges consist of (TTAGGG)_n, which is identical to the sequence type found in many higher multicellular animals and in Mastigophora. Additionally, this is the first study to reveal a telomere sequence type for Porifera. Moreover, these results suggest that Porifera are phylogenetically related to Mastigophora, and supports the general theory that Porifera evolved from Mastigophora. Further, this study strongly suggests that the origin of the (TTAGGG)_n telomere sequence is to be found in a common ancestor of either the Bilateria and Porifera, or the Protista.     

Changes in physical properties of heat-induced gel on addition of gluconate associated with suppression of myosin denaturation in walleye pollack salt-ground <Emphasis Type="Italic">surimi</Emphasis> during preheating

Changes in physical properties of two-step heated gels on addition of gluconate were investigated in terms of relationships between breaking strength and gel stiffness. Regression lines between the breaking strength and the gel stiffness were extended to the x-axis (gel stiffness), and the intercept was defined as S_BSO. The S_BSO of the two-step heated gels increased with gluconate contents in salt-ground surimis, suggesting that the harder but less elastic gels formed on addition of gluconate were dose-dependent. Conversely, the denaturation rate constants of myosin in salt-ground surimis during preheating estimated by means of Ca-ATPase inactivation, loss of salt solubility, and decrease of denaturant solubility were considerably reduced by gluconate. Thus, the progress of myosin denaturation was strongly suppressed. Increments of S_BSO (δS_BSO) of the two-step heated gels on addition of gluconate were inversely correlated with the denaturation rate constants of myosin in salt-ground surimis for every index. Thus, the changes in physical parameters of two-step heated gel caused by gluconate may be associated with the sluggish progress of myosin denaturation in salt-ground surimi during preheating.     

Highly efficient induction of triploid Pacific abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> by caffeine treatment

ABSTRACT:   In order to develop a highly efficient method for mass production of triploid Pacific abalone Haliotis discus hannai , caffeine treatment that is safe and inexpensive was optimized. To suppress the first meiotic division, fertilized eggs were exposed to either a 10- or 15-mM caffeine solution for 24 min beginning at 12 min after fertilization. In most treated batches, the rates of cleaved eggs showed no significant difference from the control batches. However, in most treated batches, the rates of occurrence of normal larvae and the survival rates of the early juveniles were significantly lower than those of the controls. The triploid rates at 6 days to 11 months after settlement in all the treated batches were extremely high (91–100%). There was no significant difference in the mean triploid rates between 10- and 15-mM caffeine treatments. These results suggest that both treatment conditions were conducive to triploid abalone production. One live 2n/3n mosaic specimen was found in the treated batches. However, since the frequency of mosaic was extremely low, the mosaicism would probably not have an adverse effect on the stable production of triploid abalones.     

Determination of genome size of Haliotis discus hannai and H.?diversicolor aquatilis (Haliotidae) and phylogenetic examination of this family

Genome size (C value) is an important index for phylogenetic studies. Haliotidae (abalones) includes many species widely distributed throughout the world??s oceans, which makes this family interesting for phylogenetic studies. To examine Haliotidae phylogeny, we determined the C value and adenine and thymine base pair content (AT?%) of Haliotis discus hannai and H.?diversicolor aquatilis by using flow cytometry. The C values of H.?discus hannai and H.?diversicolor aquatilis were 1.84 and 1.45?pg, with AT?% of 62.3 and 66.3?%, respectively. These data represent the first report of abalones classified as Pacific Northwest (H.?discus hannai) and Indo-Pacific (H.?diversicolor aquatilis) groups, and provides new validation for previous theories related to Haliotidae phylogeny.     

Effects of smolt size and timing of migration on recovery rate of wild masu salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis>

Little is known about the survival rate of wild masu salmon Oncorhynchus masou. To examine the effects of smolt length and migration timing on the recovery rate of wild masu salmon, we reanalyzed past tagging and recovery data (1993–1994). The tagging study was conducted in the Shokanbetsu River, northern Japan; 863 wild masu salmon smolts were captured, tagged, and released in a downstream site, and a total of 19 fish were recovered in coastal fisheries and in the natal river the following year. The data were analyzed by a logistic regression analysis with recapture as a response variable and tagging date and smolt length as explanatory variables; the tagging date had a significant effect on the recapture rate, whereas the effect of smolt size was not significant. Despite the small number of recaptures, this study indicates that migration timing is a factor affecting the marine survival of wild masu salmon smolts, although this conclusion has been repeatedly documented for other species of Pacific salmon Oncorhynchus spp.     

Comparison of the myocardial performance index derived by use of pulsed Doppler echocardiography and tissue Doppler imaging in dogs with volume overload

OBJECTIVE: To investigate the relationship between the myocardial performance index (MPI) determined by use of pulsed Doppler (PD) echocardiography and tissue Doppler imaging (TDI) in the response to volume overload-related changes in left ventricle (LV) performance. ANIMALS: 7 male Beagles. PROCEDURES: Dogs were anesthetized and intubated. A 6-F fluid-filled catheter was placed in the LV to measure LV peak systolic (LVPs) and LV end-diastolic (LVED) pressures. Preload was increased by IV infusion of lactated Ringer's solution (rate of 200 mL/kg/h for 60 minutes) into a cephalic vein. Transmitral flow velocities and aortic outflow were measured, and TDI velocities were obtained from the 4-chamber view. RESULTS: Acute volume overload induced a significant increase in heart rate, LVPs pressure, and LVED pressure, compared with baseline values. A significant decrease in the PD-MPI and TDI-MPI values and a significant correlation (r = 0.70) between PD-MPI and TDI-MPI were detected. The PD-derived A-wave velocity, ejection time, and isovolumic relaxation time (IRT) and the TDI-derived IRT, MPI, and ratio of the velocity of the E wave to the velocity of the ventricular portion of the E wave during early diastole had equal ability to predict LVED pressure (r(2) = 0.63). CONCLUSIONS AND CLINICAL RELEVANCE: The TDI-MPI was closely correlated with LV filling pressure and may be helpful in evaluating global cardiac function in dogs.