The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration

Integrin β3 is seen as a key anti‐angiogenic target for cancer treatment due to its expression on neovasculature, but the role it plays in the process is complex; whether it is pro‐ or anti‐angiogenic depends on the context in which it is expressed. To understand precisely β3's role in regulating integrin adhesion complexes in endothelial cells, we characterised, by mass spectrometry, the β3‐dependent adhesome. We show that depletion of β3‐integrin in this cell type leads to changes in microtubule behaviour that control cell migration. β3‐integrin regulates microtubule stability in endothelial cells through Rcc2/Anxa2‐driven control of active Rac1 localisation. Our findings reveal that angiogenic processes, both in vitro and in vivo, are more sensitive to microtubule targeting agents when β3‐integrin levels are reduced.     

Spontaneous activation of fish eggs is abolished by protease inhibitors

During spawning, eggs of most fish species entering the aquatic environment remain fertilizable for a relatively short period of time. This is due to the “spontaneous egg activation” giving rise to the fertilization membrane, which prevents the penetration of excessive and foreign sperm into the egg during normal fertilization. This work demonstrates that the fertilization membrane formation and the loss of fertilizability in aqueous solutions of different composition are inhibited by protease inhibitors, in particular, leupeptin and aprotinin. The presence of natural protease inhibitors in the ovarian fluid that prevent spontaneous egg activation is proposed. The decrease in the concentration of these inhibitors as the ovarian fluid is diluted in aquatic medium during spawning can explain egg activation in the absence of sperm.     

The effect of local cortical microfilament disorganization on ooplasmic segregation in the loach (Misgurnus fossilis) egg

Injections of cytochalasin D (CD) or DNase I under the surface of fertilized loach egg result in local disorganization of microfilamentous cortex (MC) as revealed by transmission electron microscopy. This effect correlates with the loss of the cortex ability to contract in vitro. The disorganization of MC in the vegetal hemisphere of the egg does not affect the ooplasm segregation or blastodisk cleavage. Injection under the animal pole suppresses blastodisk formation and results in the autonomous separation of ooplasm in the central part of the egg. The experiments suggest that (1) autonomous separation of ooplasm from the yolk granules can proceed in the central part of the egg without the participation of MC; (2) normal segregation of ooplasm at the animal pole requires that the structures of microfilaments in the animal hemisphere (but not in the vegetal one) be preserved.     

Nitric oxide synthase is up-regulated in muscle fibers in muscular dystrophy

Nitric oxide (NO) mediates fundamental physiological actions on skeletal muscle. The neuronal NO synthase isoform (NOS1) was reported to be located exclusively in the sarcolemma. Its loss from the sarcolemma was associated with development of Duchenne muscular dystrophy (DMD). However, new studies evidence that all three NOS isoforms-NOS1, NOS2, and NOS3-are co-expressed in the sarcoplasm both in normal and in DMD skeletal muscles. To address this controversy, we assayed NOS expression in DMD myofibers in situ cytophotometrically and found NOS expression in DMD myofibers up-regulated. These results support the hypothesis that NO deficiency with consequent muscle degeneration in DMD results from NO scavenging by superoxides rather than from reduced NOS expression.     

The role of endogenous glutathione in the action of sulfur-containing radio-protectors

In experiments on Ehrlich ascite tumor cells, the dependence of the radioprotective effect of beta-mercaptoethylamine and beta-mercaptopropionylglycine on the initial level of endogenous glutathione was studied. A varying degree of N-ethylmaleimide induced decrease of SH-glutathione content in the cells led to either easing or elimination of the radioprotective effect of the agents under study.     

Influence of primary adhesive interactions with fibronectin on clonal growth and osteogenic potential of rat mesenchymal stromal cells

Interactions with the extracellular matrix play an important role in the regulation of cell growth and differentiation. Fibronectin (Fn) is a major component of the extracellular matrix. The impact of Fn and its domains on the adhesion and osteogenic potency of rat mesenchymal stromal cells (MSC) have been evaluated. The examination of bone marrow and fetal liver MSC adhesion dynamics showed that after 7 days of cultivation on Fn the number of adhered clonogenic cells derived from both sources was comparable with their number on plastic but their content in suspension usually was decreased. The population of fetal liver MSC differed from bone marrow-derived MSC in the larger number of cells that adhered for the first 7 days. We found that the primary adhesion of MSC to Fn affected their differentiation potency. Bone marrow MSC cultured on Fn had reduced activity of alkaline phosphatase compared with cells grown on plastic. They deposited a significantly less amount of calcium salts in osteogenic medium. The cultivation of MSC on Fn fragments demonstrated that the crucial role in the inhibition of osteogenesis is played by the Fn cell-binding domain.