Myosin VI small insert isoform maintains exocytosis by tethering secretory granules to the cortical actin.

Before undergoing neuroexocytosis, secretory granules (SGs) are mobilized and tethered to the cortical actin network by an unknown mechanism. Using an SG pull-down assay and mass spectrometry, we found that myosin VI was recruited to SGs in a Ca(2+)-dependent manner. Interfering with myosin VI function in PC12 cells reduced the density of SGs near the plasma membrane without affecting their biogenesis. Myosin VI knockdown selectively impaired a late phase of exocytosis, consistent with a replenishment defect. This exocytic defect was selectively rescued by expression of the myosin VI small insert (SI) isoform, which efficiently tethered SGs to the cortical actin network. These myosin VI SI-specific effects were prevented by deletion of a c-Src kinase phosphorylation DYD motif, identified in silico. Myosin VI SI thus recruits SGs to the cortical actin network, potentially via c-Src phosphorylation, thereby maintaining an active pool of SGs near the plasma membrane

Study on the effects of stand density management of Chinese fir plantation in Northern China

The aim of this study was to clarify the mechanism by which thinning alters stand structure and affects forest productivity by characterizing changes in stand quantitative maturity age, stand diameter distribution, structural heterogeneity, and forest productivity of Chinese fir plantations at different thinning times and intensities. Our findings provide insights into how the density of stands could be modified to enhance the yield and timber quality of Chinese fir plantations. The significance of differences in individual tree volume, stand volume, and timber merchantable volume was determined using one-way analysis of variance and post hoc Duncan tests. The stand quantitative maturity age was obtained using the Richards equation. The quantitative relationship between stand structure and productivity was determined using a generalized linear mixed model. We found that (1) the quantitative maturity age of Chinese fir plantations increased with thinning intensity, and the quantitative maturity age was much greater under commercial thinning than under pre-commercial thinning. (2) Individual tree volume and the proportion of medium-sized and large-sized timber merchantable volume increased with stand thinning intensity. Thinning promoted increases in stand diameter. pre-commercially thinned stands were dominated by medium-diameter trees when the quantitative maturity age was reached, whereas commercially thinned stands were dominated by large-diameter trees. The living trees volume will decrease immediately after thinning, and then it will gradually increase with the age of the stand. When the stand volume included both living trees volume and thinned volume, thinned stands increased stand volume compared with unthinned stands. In pre-commercial thinning stands, the greater the intensity of thinning, the greater the increase in stand volume, and the opposite was true for commercial thinning. (3) Thinning also reduced heterogeneity in stand structure, which was lower after commercial thinning than after pre-commercial thinning. The productivity of pre-commercially thinned stands increased with thinning intensity, whereas that of commercially thinned stands decreased with thinning intensity. (4) The structural heterogeneity of pre-commercially and commercially thinned stands was negatively and positively correlated with forest productivity, respectively. In the Chinese fir plantations in the hilly terrain of the northern Chinese fir production area, when pre-commercial thinning was performed in the ninth year to a residual density of 1750 trees per hectare, the stand quantitative maturity age was reached in year 30, medium-sized timber accounted for 75.2% of all trees, and the stand volume was 667.9 m3 per hectare. This thinning strategy is favorable for producing medium-sized Chinese fir timber. When commercial thinning was performed in year 23, the optimal residual density was 400 trees per hectare. When the stand quantitative maturity age was reached in year 31, large-sized timber accounted for 76.6% of all trees, and the stand volume was 574.5 m3 per hectare. This thinning strategy is favorable for producing large-sized Chinese fir timber

Expression of scinderin in megakaryoblastic leukemia cells induces differentiation, maturation, and apoptosis with release of plateletlike particles and inhibits proliferation and tumorigenesis

Rapid proliferation of atypical megakaryoblasts is a characteristic of megakaryoblastic leukemia. Cells from patients with this disorder and cell lines established from this type of leukemia showed the presence of gelsolin but the absence of scinderin expression, 2 filamentous actin-severing proteins present in normal megakaryocytes and platelets. Vector-mediated expression of scinderin in the megakaryoblastic cell line MEG-01 induced a decrease in both F-actin and gelsolin. This was accompanied by increased Rac2 expression and by activation of the PAK/MEKK.SEK/JNK/c-jun, c-fos transduction pathway. The Raf/MEK/ERK pathway was also activated in these cells. Transduction pathway activation was followed by cell differentiation, polyploidization, maturation, and apoptosis with release of platelet-like particles. Particles expressed surface CD41a antigen (glycoprotein IIb/IIIa or fibrinogen receptor), had dense bodies, high-affinity serotonin transport, and circular array of microtubules. Treatment of particles with thrombin induced serotonin release and aggregation that was blocked by CD41a antibodies. PAC-1 antibodies also blocked aggregation. Exposure of cells to PD98059, a blocker of MEK, inhibited antigen CD41a expression, increases in cell volume, and number of protoplasmic extensions. Cell proliferation and cell ability to form tumors in nude mice were also inhibited by the expression of scinderin. MEG-01 cells expressing scinderin had the same fate in vivo as in culture. Thus, when injected into nude mice, they entered apoptosis and released platelet-like particles. The lack of scinderin expression in megakaryoblastic leukemia cells seems to be responsible for their inability to enter into differentiation and maturation pathways characteristic of their normal counterparts. © 2001 by The American Society of Hematology.Fil: Zunino, Rodolfo. University of Ottawa; CanadáFil: Li, Qinggang. University of Ottawa; CanadáFil: Rosé, Sergio Daniel. University of Ottawa; CanadáFil: Romero-Benítez, María Margarita Itatí. University of Ottawa; CanadáFil: Lejen, Tatiana. University of Ottawa; CanadáFil: Brandan, Nora Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Nordeste. Facultad de Medicina; Argentina. University of Ottawa; CanadáFil: Trifaró, José-María. University of Ottawa; Canad