Release of vasopressin from isolated permeabilized neurosecretory nerve terminals is blocked by the light chain of botulinum A toxin

The intracellular action on exocytosis of botulinim A toxin and constituent chains was studied using permeabilized isolated nerve endings from the rat neural lobe. The release of the neuropeptide vasopressin was measured by radioimmunoassay. In the presence of the reducing agent dithiothreitol, the two-chain form of botulinum A toxin inhibited vasopressin release induced by 10 μM free calcium. Half maximal inhibition was obtained with 15 nM botulinum A toxin. In the absence of the heavy chain the light chain of the toxin strongly inhibited exocytosis with a half maximal effect of 2.5 nM. The inhibitory effects on secretion could be prevented by incubating the light chain with an immune serum against botulinum A toxin. The heavy chain of botulinum A toxin did not affect vasopressin release. However, it prevented the inhibitory effects of the light chain on stimulated exocytosis. It is concluded that botulinum A toxin inhibits the calcium-dependent step leading to exocytosis by interfering with a target present in the isolated and permeabilized nerve terminals. The functional domain of this neurotoxin, which is responsible for the inhibition of vasopressin release, is present in its light chain

Release of vasopressin from isolated permeabilized neurosecretory nerve terminals is blocked by the light chain of botulinum A toxin

The intracellular action on exocytosis of botulinim A toxin and constituent chains was studied using permeabilized isolated nerve endings from the rat neural lobe. The release of the neuropeptide vasopressin was measured by radioimmunoassay. In the presence of the reducing agent dithiothreitol, the two-chain form of botulinum A toxin inhibited vasopressin release induced by 10 μM free calcium. Half maximal inhibition was obtained with 15 nM botulinum A toxin. In the absence of the heavy chain the light chain of the toxin strongly inhibited exocytosis with a half maximal effect of 2.5 nM. The inhibitory effects on secretion could be prevented by incubating the light chain with an immune serum against botulinum A toxin. The heavy chain of botulinum A toxin did not affect vasopressin release. However, it prevented the inhibitory effects of the light chain on stimulated exocytosis. It is concluded that botulinum A toxin inhibits the calcium-dependent step leading to exocytosis by interfering with a target present in the isolated and permeabilized nerve terminals. The functional domain of this neurotoxin, which is responsible for the inhibition of vasopressin release, is present in its light chain

The light chain of tetanus toxin inhibits calcium-dependent vasopressin release from permeabilized nerve endings

The effects of tetanus toxin and its light and heavy chain subunits on vasopressin release were investigated in digitonin-permeabilized neurosecretory nerve terminals isolated from the neural lobe of the rat pituitary gland. Exocytosis was induced by challenging the permeabilized nerve endings with micromolar calcium concentrations. Tetanus toxin inhibited vasopressin release only in the presence of the reducing agent dithiothreitol. This effect was irreversible. The purified light chain of tetanus toxin strongly inhibited exocytosis in a dose-dependent manner with half-maximal effect at c. 10 nM. The action of the light chain was observed after only 2.5 min of preincubation. Separated heavy chain subunit had no effect on hormone secretion. Inhibition of vasopressin release could be prevented by preincubating the light chain of tetanus toxin with an immune serum against tetanus toxin. The data clearly demonstrate that in mammalian neurosecretory nerve endings tetanus toxin acts at a step downstream from the activation by Ca2+ of the exocytotic machinery and that the functional domain of this toxin is confined to its light chain

GTP and Ca2+ Modulate the Inositol 1,4,5-Trisphosphate-Dependent Ca2+ Release in Streptolysin O-Permeabilized Bovine Adrenal Chromaffin Cells

The inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ release was studied using streptolysin O-permeabilized bovine adrenal chromaffin cells. The IP3-induced Ca2+ release was followed by Ca2+ reuptake into intracellular compartments. The IP3-induced Ca2+ release diminished after sequential applications of the same amount of IP3. Addition of 20 μM GTP fully restored the sensitivity to IP3. Guanosine 5'-O-(3-thio)triphosphate (GTPγS) could not replace GTP but prevented the action of GTP. The effects of GTP and GTPγS were reversible. Neither GTP nor GTPγS induced release of Ca2+ in the absence of IP3. The amount of Ca2+ whose release was induced by IP3 depended on the free Ca2+ concentration of the medium. At 0.3 μM free Ca2+, a half-maximal Ca2+ release was elicited with ∼0.1 μM IP3. At 1 μM free Ca2+, no Ca2+ release was observed with 0.1 μM IP3; at this Ca2+ concentration, higher concentrations of IP3 (0.25 μM) were required to evoke Ca2+ release. At 8 μM free Ca2+, even 0.25 μM IP3 failed to induce release of Ca2+ from the store. The IP3-induced Ca2+ release at constant low (0.2 μM) free Ca2+ concentrations correlated directly with the amount of stored Ca2+. Depending on the filling state of the intracellular compartment, 1 mol of IP3 induced release of between 5 and 30 mol of Ca2+

Chiral QCD sum rules for open charm mesons

QCD sum rules for chiral partners in the open-charm meson sector are presented at nonzero baryon net density or temperature. We focus on the differences between pseudo-scalar and scalar as well as vector and axial-vector D mesons and derive the corresponding Weinberg type sum rules. This allows for the identification of such QCD condensates which drive the non-degeneracy of chiral partners in lowest order of the strong coupling alpha_s and which therefore may serve as "order parameters" for chiral restoration (or elements thereof).Comment: 24 pages, 4 figure

Introduction of Macromolecules into Bovine Adrenal Medullary Chromaffin Cells and Rat Pheochromocytoma Cells (PC12) by Permeabilization with Streptolysin O: Inhibitory Effect of Tetanus Toxin on Catecholamine Secretion

Conditions are described for controlled plasma membrane permeabilization of rat pheochromocytoma cells (PC12) and cultured bovine adrenal chromaffin cells by Streptolysin O (SLO). The transmembrane pores created by SLO invoke rapid efflux of intracellular 86Rb+ and ATP, and also permit passive diffusion of proteins, including immunoglobulins, into the cells. SLO-permeabilized PC12 cells release [3H]dopamine in response to micromolar concentrations of free Ca2+. Permeabilized adrenal chromaffin cells present a similar exocytotic response to Ca2+ in the presence of Mg2+/ ATP. Permeabilized PC12 cells accumulate antibodies against synaptophysin and calmodulin, but neither antibody reduces the Ca2+-dependent secretory response. Reduced tetanus toxin, although ineffective when applied to intact chromaffin cells, inhibits Ca2+-induced exocytosis by both types of permeabilized cells studied. Omission of dithiothreitol, toxin inactivation by boiling, or preincubation with neutralizing antibodies abolishes the inhibitory effect. The data indicate that plasma membrane permeabilization by Streptolysin O is a useful tool to probe and define cellular components that are involved in the final steps of exocytosis

R-matrix approach to integrable systems on time scales

A general unifying framework for integrable soliton-like systems on time scales is introduced. The $R$-matrix formalism is applied to the algebra of $\delta$-differential operators in terms of which one can construct infinite hierarchy of commuting vector fields. The theory is illustrated by two infinite-field integrable hierarchies on time scales which are difference counterparts of KP and mKP. The difference counterparts of AKNS and Kaup-Broer soliton systems are constructed as related finite-field restrictions.Comment: 21 page

surface interpolation and 3d relatability

Although the role of surface-level processes has been demonstrated, visual interpolation models often emphasize contour relationships. We report two experiments on geometric constraints governing 3D interpolation between surface patches without visible edges. Observers were asked to classify pairs of planar patches specified by random dot disparities and visible through circular apertures (aligned or misaligned) in a frontoparallel occluder. On each trial, surfaces appeared in parallel or converging planes with vertical (in Experiment 1) or horizontal (in Experiment 2) tilt and variable amounts of slant. We expected the classification task to be facilitated when patches were perceived as connected. We found enhanced sensitivity and speed for 3D relatable vs. nonrelatable patches. Here 3D relatability does not involve oriented edges but rather inducing patches' orientations computed from stereoscopic information. Performance was markedly affected by slant anisotropy: both sensitivity and speed were worse for patches with horizontal tilt. We found nearly identical advantages of 3D relatability on performance, suggesting an isotropic unit formation process. Results are interpreted as evidence that inducing slant constrains surface interpolation in the absence of explicit edge information: 3D contour and surface interpolation processes share common geometric constraints as formalized by 3D relatability

Release of vasopressin from isolated permeabilized neurosecretory nerve terminals is blocked by the light chain of botulinum A toxin

The intracellular action on exocytosis of botulinim A toxin and constituent chains was studied using permeabilized isolated nerve endings from the rat neural lobe. The release of the neuropeptide vasopressin was measured by radioimmunoassay. In the presence of the reducing agent dithiothreitol, the two-chain form of botulinum A toxin inhibited vasopressin release induced by 10 μM free calcium. Half maximal inhibition was obtained with 15 nM botulinum A toxin. In the absence of the heavy chain the light chain of the toxin strongly inhibited exocytosis with a half maximal effect of 2.5 nM. The inhibitory effects on secretion could be prevented by incubating the light chain with an immune serum against botulinum A toxin. The heavy chain of botulinum A toxin did not affect vasopressin release. However, it prevented the inhibitory effects of the light chain on stimulated exocytosis. It is concluded that botulinum A toxin inhibits the calcium-dependent step leading to exocytosis by interfering with a target present in the isolated and permeabilized nerve terminals. The functional domain of this neurotoxin, which is responsible for the inhibition of vasopressin release, is present in its light chain

Assessing a Hydrodynamic Description for Instabilities in Highly Dissipative, Freely Cooling Granular Gases

An intriguing phenomenon displayed by granular flows and predicted by kinetic-theory-based models is the instability known as particle "clustering," which refers to the tendency of dissipative grains to form transient, loose regions of relatively high concentration. In this work, we assess a modified-Sonine approximation recently proposed [Garz\'o et al., Physica A 376, 94 (2007)] for a granular gas via an examination of system stability. In particular, we determine the critical length scale associated with the onset of two types of instabilities -vortices and clusters- via stability analyses of the Navier-Stokes-order hydrodynamic equations by using the expressions of the transport coefficients obtained from both the standard and the modified-Sonine approximations. We examine the impact of both Sonine approximations over a range of solids fraction \phi <0.2 for small restitution coefficients e=0.25--0.4, where the standard and modified theories exhibit discrepancies. The theoretical predictions for the critical length scales are compared to molecular dynamics (MD) simulations, of which a small percentage were not considered due to inelastic collapse. Results show excellent quantitative agreement between MD and the modified-Sonine theory, while the standard theory loses accuracy for this highly dissipative parameter space. The modified theory also remedies a (highdissipation) qualitative mismatch between the standard theory and MD for the instability that forms more readily. Furthermore, the evolution of cluster size is briefly examined via MD, indicating that domain-size clusters may remain stable or halve in size, depending on system parameters.Comment: 4 figures; to be published in Phys. Rev.