Optimum Calcium Concentration: A Crucial Factor in Regulating Sperm Motility In Vitro

Sperm motility can be maintained in vitro by incubation in a defined medium under specific conditions. In most studies, the exact role of various constituents of epididymal fluid, including calcium, has remained obscure. Most of the culture media have included millimolar concentrations of calcium, but previous reports have indicated that millimolar calcium inhibits sperm motility. In this present study, we sought the optimum concentration of extracellular calcium required for optimum sperm motility. This study showed that extracellular calcium has a concentration-dependent biphasic role in motility regulation. It promoted motility and velocity at lower (10 µM) concentration whereas notably inhibited it at higher concentrations. When external membrane-bound calcium was removed by ethylene glycol tetraacetic acid, motility decreased considerably. To confirm the motility-inhibiting role of calcium above 10 µM, a sperm motility-stimulating protein (MSP) recently reported from our laboratory was used which at 0.9 μM induces motility in 60–70 % cells. Calcium at 10 µM had no appreciable effect on the motility-promoting activity of the MSP but depressed the activity above 10 µM. Thus, our present results emphasize the biphasic role of extracellular calcium and the importance of its optimum concentration in different buffers and media used for sperm motility initiation.     

FLJ25439, a novel cytokinesis-associated protein,induces tetraploidization and maintains chromosomal stability via enhancing expression of endoplasmic reticulum stress chaperones

Investigation of the mechanisms leading to aneuploidy and polyploidy is critical to cancer research. Previous studies have provided strong evidence of the importance of tetraploidization as an early step in tumorigenesis. In cancer cells, tetraploid cells may contribute to abnormal mitotic progression, which may be associated with cytokinesis failure. Tetraploidy leads to genomic instability due to centrosome and chromosome over-replication. Until now, the mechanism by which cells maintain tetraploid status has been unknown. Here, we identified a novel D box-containing protein, FLJ25439, which displays a dynamic expression profile during mitosis/cytokinesis with the midbody as the most prominent associated structure. To understand the function of FLJ25439, we established stable cell lines overexpressing FLJ25439. FLJ25439-overexpression cells grew slower and displayed a tetraploid DNA content in comparison with diploid parental cells. They also showed aberrant mitosis and dysregulated expression of p53, pRb and p21, suggesting a defect in cell cycle progression. To explore the molecular mechanisms responsible for FLJ25439-induced tetraploidization, we conducted a comparative analysis of the global protein expression patterns of wild type and overexpressors using proteomics and bioinformatics approaches. Protein category profiling indicated that FLJ25439 is involved in pathways related to anti-apoptosis, protein folding, the cell cycle, and cytoskeleton regulation. Specifically, genotoxic-stress- and ER stress-related chaperone proteins greatly contributed to the FLJ25439 overexpression phenotypes. The results of this study pave the way to our further understanding of the role of this novel cytokinesis-related protein in protecting cells from environmental stress and tetraploid formation.     

Nanoceria inhibit the development and promote the regression of pathologic retinal neovascularization in the Vldlr knockout mouse

Many neurodegenerative diseases are known to occur and progress because of oxidative stress, the presence of reactive oxygen species (ROS) in excess of the cellular defensive capabilities. Age related macular degeneration (AMD), diabetic retinopathy (DR) and inherited retinal degeneration share oxidative stress as a common node upstream of the blinding effects of these diseases. Knockout of the Vldlr gene results in a mouse that develops intraretinal and subretinal neovascular lesions within the first month of age and is an excellent model for a form of AMD called retinal angiomatous proliferation (RAP). Cerium oxide nanoparticles (nanoceria) catalytically scavenge ROS by mimicking the activities of superoxide dismutase and catalase. A single intravitreal injection of nanoceria into the Vldlr-/- eye was shown to inhibit: the rise in ROS in the Vldlr-/- retina, increases in vascular endothelial growth factor (VEGF) in the photoreceptor layer, and the formation of intraretinal and subretinal neovascular lesions. Of more therapeutic interest, injection of nanoceria into older mice (postnatal day 28) resulted in the regression of existing vascular lesions indicating that the pathologic neovessels require the continual production of excessive ROS. Our data demonstrate the unique ability of nanoceria to prevent downstream effects of oxidative stress in vivo and support their therapeutic potential for treatment of neurodegenerative diseases such as AMD and DR.     

Glycine rich proline rich protein from Sorghum bicolor serves as an antimicrobial protein implicated in plant defense response

Plant Molecular Biology - That overexpression of GmKR3 enhances innate virus resistance by stimulating. Soybean mosaic virus (SMV) is found in many soybean production areas, and SMV infection is...