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Oregon Green (oregon + green)
Selected AbstractsSevering of F-actin by yeast cofilin is pH-independentCYTOSKELETON, Issue 9 2006Dmitry Pavlov Abstract Cofilin plays an important role in actin turnover in cells by severing actin filaments and accelerating their depolymerization. The role of pH in the severing by cofilin was examined using fluorescence microscopy. To facilitate the imaging of actin filaments and to avoid the use of rhodamine phalloidin, which competes with cofilin, ,-actin was labeled with tetramethylrhodamine cadaverine (TRC) at Gln41. The TRC-labeling inhibited actin treadmilling strongly, as measured by ,ATP release. Cofilin binding, detected via an increase in light scattering, and the subsequent conformational change in filament structure, as detected by TRC fluorescence decay, occurred 2,3 times faster at pH 6.8 than at pH 8.0. In contrast, actin filaments severing by cofilin was pH-independent. The pH-independent severing by cofilin was confirmed using actin labeled at Cys374 with Oregon Green® 488 maleimide. The depolymerization of actin by cofilin was faster at high pH. Cell Motil. Cytoskeleton 2006. © 2006 Wiley-Liss, Inc. [source] Temperature gradient focusing in a PDMS/glass hybrid microfluidic chipELECTROPHORESIS, Issue 24 2007Takuya Matsui Abstract This paper reports the application of temperature gradient focusing (TGF) in a PDMS/glass hybrid microfluidic chip. With TGF, by the combination of a temperature gradient along a microchannel, an applied electric field, and a buffer with a temperature-dependent ionic strength, analytes are focused by balancing their electrophoretic velocities against the bulk velocity of the buffer containing the analytes. In this work, Oregon Green 488 carboxylic acid was concentrated approximately 30 times as high as the initial concentration in 45,s at moderate electric strength of 70,V/cm and a temperature gradient of 55°C across the PDMS/glass hybrid microfluidic chip with a 1,cm long capillary. [source] No evidence for calcium electrogenic exchanger in frog semicircular canal hair cellsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2002M. Martini Abstract We investigated the possibility that, in hair cells mechanically isolated from frog semicircular canals, Ca2+ extrusion occurs via a Na+ : Ca2+ (cardiac type) or a Na+ : Ca2+,K+ (retinal type) exchanger. Cells concurrently imaged during whole-cell patch-clamp recordings using the Ca2+ sensitive fluorescent dye Oregon Green 488 BAPTA-1 (100 µm) showed no voltage dependence of Ca2+ clearance dynamics following a Ca2+ load through voltage-gated Ca2+ channels. Reverse exchange was probed in hair cells dialyzed with a Ca2+ - and K+ -free solution, containing a Na+ concentration that saturates the exchanger, after zeroing the contribution to the whole-cell current from Ca2+ and K+ conductances. In these conditions, no reverse exchange current was detected upon switching from a Ca2+ -free external solution to a solution containing concentrations of Ca2+ alone, or Ca2+ + K+ that saturated the exchanger. By contrast, the same experimental protocol elicited peak exchange currents exceeding 100 pA in gecko rod photoreceptors, used as positive controls. In both cell types, we also probed the forward mode of the exchanger by rapidly increasing the intracellular Ca2+ concentration using flash photolysis of two novel caged Ca2+ complexes, calcium 2,2,-{[1-(2-nitrophenyl)ethane-1,2-diyl]bis(oxy)}bis(acetate) and calcium 2,2,-{[1-(4,5-dimethoxy-2-nitrophenyl)ethane-1,2-diyl]bis(oxy)} bis(acetate), in the presence of internal K+ and external Na+. No currents were evoked by UV-triggered Ca2+ jumps in hair cells, whereas exchanger conformational currents up to 400 pA, followed by saturating forward exchange currents up to 40 pA, were recorded in rod photoreceptors subjected to the same experimental conditions. We conclude that no functional electrogenic exchanger is present in this hair cell population, which leaves the abundant plasma membrane Ca2+ -ATPases as the primary contributors to Ca2+ extrusion. [source] Ni2+ induces changes in the morphology of vacuoles, mitochondria and microtubules in Paxillus involutus cellsNEW PHYTOLOGIST, Issue 4 2006Sandra Tuszy Summary ,,Organelles of ectomycorrhizal fungi are known to respond to changes in the extracellular environment. The response of vacuoles, mitochondria and microtubules to short-term nickel (Ni2+) exposure were investigated in hyphal tip cells of a Paxillus involutus from a heavy metal-rich soil. ,,Vacuoles, mitochondria and microtubules were labelled with Oregon Green® 488 carboxylic acid diacetate, 3,3,-dihexyloxacarbocyanine iodide (DiOC6(3)) and anti-,-tubulin antibodies, respectively; hyphae were treated with NiSO4 in the range of 0,1 mmol l,1 and examined microscopically. ,,Untreated hyphal tip cells contained tubular vacuole and mitochondrial networks. Ni2+ caused loss of organelle tubularity and severe microtubule disruption that were exposure-time and concentration dependent. Fine tubular vacuoles thickened and eventually became spherical in some hyphae, tubular mitochondria fragmented and microtubules shortened and aggregated into patches in most hyphae. Tubular vacuoles reformed on NiSO4 removal and tubular mitochondria in the presence of NiSO4 suggesting cellular detoxification. ,,These results demonstrate that Ni2+ induces changes in organelle and microtubule morphology. Recovery of tubular organelles to pretreatment morphology after Ni2+ exposure suggests cellular detoxification of the metal ion. [source] Visualisation of the uptake of two model xenobiotics into bean leaves by confocal laser scanning microscopy: diffusion pathways and implication in phloem translocationPEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 5 2004Dr Zhiqian Liu Abstract The diffusion of two fluorescent dyes, Oregon Green 488 (Oregon Green) and Rhodamine B into the leaves of broad bean (Vicia faba L) plants was studied to simulate the foliar uptake process of pesticides. The uptake rate of these model xenobiotics into bean foliage was measured using a standard leaf surface wash-off method. Diffusion into leaf tissues was visualised in vivo by confocal laser scanning microscopy (CLSM). The moderately lipophilic dye (Rhodamine B) showed faster uptake than the hydrophilic one (Oregon Green), despite the former being a larger molecule. While no distinct channels or domains for preferential entry of any of the dyes could be detected in the cuticle layer by CLSM, two different diffusion patterns were identified for the movement of these two dyes after traversing the cuticle. Upon desorption from the cuticle, Rhodamine B diffused extensively into the vacuole of the epidermal cells. Further transport of this dye from the epidermal cells to the mesophyll cells was not observed. In contrast, Oregon Green was found in the epidermal cell walls and cytoplasm, and was also present in the mesophyll cells. Examination of the petioles of the treated leaves revealed that, once absorbed, Oregon Green moved readily out of the treated leaf, whereas Rhodamine B did not show any phloem translocation. It is proposed that these two different diffusion characters may be responsible for the contrasting phloem mobility of the two xenobiotics. The results are discussed in relation to the current knowledge on the uptake, translocation and efficacy of pesticides as influenced by their properties. Copyright © 2004 Society of Chemical Industry [source] | ||||||||||