Peri/epicellular protein disulfide isomerase-A1 acts as an upstream organizer of cytoskeletal mechanoadaptation in vascular smooth muscle cells.

Although redox processes closely interplay with mechanoresponses to control vascular remodeling, redox pathways coupling mechanostimulation to cellular cytoskeletal organization remain unclear. The peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) supports postinjury vessel remodeling. Using distinct models, we investigated whether pecPDIA1 could work as a redox-dependent organizer of cytoskeletal mechanoresponses. In vascular smooth muscle cells (VSMCs), pecPDIA1 immunoneutralization impaired stress fiber assembly in response to equibiaxial stretch and, under uniaxial stretch, significantly perturbed cell repositioning perpendicularly to stretch orientation. During cyclic stretch, pecPDIA1 supported thiol oxidation of the known mechanosensor ß1-integrin and promoted polarized compartmentalization of sulfenylated proteins. Using traction force microscopy, we showed that pecPDIA1 organizes intracellular force distribution. The net contractile moment ratio of platelet-derived growth factor-exposed to basal VSMCs decreased from 0.90 ± 0.09 (IgG-exposed controls) to 0.70 ± 0.08 after pecPDI neutralization ( P < 0.05), together with an enhanced coefficient of variation for distribution of force modules, suggesting increased noise. Moreover, in a single cell model, pecPDIA1 neutralization impaired migration persistence without affecting total distance or velocity, whereas siRNA-mediated total PDIA1 silencing disabled all such variables of VSMC migration. Neither expression nor total activity of the master mechanotransmitter/regulator RhoA was affected by pecPDIA1 neutralization. However, cyclic stretch-induced focal distribution of membrane-bound RhoA was disrupted by pecPDI inhibition, which promoted a nonpolarized pattern of RhoA/caveolin-3 cluster colocalization. Accordingly, FRET biosensors showed that pecPDIA1 supports localized RhoA activity at cell protrusions versus perinuclear regions. Thus, pecPDI acts as a thiol redox-dependent organizer and noise reducer mechanism of cytoskeletal repositioning, oxidant generation, and localized RhoA activation during a variety of VSMC mechanoresponses. NEW & NOTEWORTHY Effects of a peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) during mechanoregulation in vascular smooth muscle cells (VSMCs) were highlighted using approaches such as equibiaxial and uniaxial stretch, random single cell migration, and traction force microscopy. pecPDIA1 regulates organization of the cytoskeleton and minimizes the noise of cell alignment, migration directionality, and persistence. pecPDIA1 mechanisms involve redox control of ß1-integrin and localized RhoA activation. pecPDIA1 acts as a novel organizer of mechanoadaptation responses in VSMCs.

Genetic structure analysis of Eufriesea violacea (Hymenoptera, Apidae) populations from southern Brazilian Atlantic rainforest remnants

Random amplified polymorphic DNA (RAPD) markers were used to analyze the genetic structure of Eufriesea violacea populations in three fragments (85.47, 832.58 and 2800 ha) of Atlantic rainforest located in the north of the Brazilian state of Paraná. A total of twelve primers produced 206 loci, of which 129 were polymorphic (95 percent criterion). The proportions of polymorphic loci in each population ranged from 57.28 percent to 59.2 percent, revealing very similar levels of genetic variability in the groups of bees from each fragment. Unbiased genetic distances between groups ranged from 0.0171 to 0.0284, the smallest genetic distance occurring between bees from the two larger fragments. These results suggest that the E. violacea populations from the three fragments have maintained themselves genetically similar to native populations of this species originally present in northern Paraná.

Euglossine bees (Hymenoptera, Apidae) in a remnant of Atlantic Forest in Paraná State, Brazil

Species composition, relative abundance, seasonal changes in the species abundance and scent association of male Euglossini collected in a semi-deciduous forest fragment in the north of the State of Paraná, southern Brazil, were recorded. Euglossine males were collected twice a month, for twelve months, from 8:00 am to 3:00 pm. The scents eucalyptol, eugenol, vanillin, methyl salicylate and benzyl acetate were used as baits. A total of 434 males distributed among 3 genera and 9 species were attracted to the chemical baits. Eufriesea violacea (Blanchard, 1840) (49.8%), Eulaema nigrita Lepeletier, 1841 (23.0%) and Euglossa pleosticta Dressler, 1982 (13.8%) were dominant in number of individuals. Among the non-dominant species, Euglossa fimbriata Rebêlo & Moure, 1995 was more common (9.0%), followed by E. cordata (L., 1758) (1.8%), E. truncata Rebêlo & Moure, 1995 (1.4%), E. melanotricha Moure, 1967 (0.7%), E. townsendi Cockerell, 1904 (0.23%) and Eufriesea auriceps Friese, 1899 (0.23%). In general, bees were more abundant in warm-wet season (September-March). Eufriesea violacea was the most seasonal species, showing activity through the warm-wet season, from October to February. Eucalyptol was the most attractive fragrance, which was responsible for 92.6% of all visits by euglossine bees.