Neural Synchronization at Tonic-to-Bursting Transitions

We studied the synchronous behavior of two electrically-coupled model neurons as a function of the coupling strength when the individual neurons are tuned to different activity patterns that ranged from tonic firing via chaotic activity to burst discharges. We observe asynchronous and various synchronous states such as out-of-phase, in-phase and almost in-phase chaotic synchronization. The highest variety of synchronous states occurs at the transition from tonic firing to chaos where the highest coupling strength is also needed for in-phase synchronization which is, essentially, facilitated towards the bursting range. This demonstrates that tuning of the neuron’s internal dynamics can have significant impact on the synchronous states especially at the physiologically relevant tonic-to-bursting transitions.     

Conductance versus current noise in a neuronal model for noisy subthreshold oscillations and related spike generation

Biological systems are notoriously noisy. Noise, therefore, also plays an important role in many models of neural impulse generation. Noise is not only introduced for more realistic simulations but also to account for cooperative effects between noisy and nonlinear dynamics. Often, this is achieved by a simple noise term in the membrane equation (current noise). However, there are ongoing discussions whether such current noise is justified or whether rather conductance noise should be introduced because it is closer to the natural origin of noise. Therefore, we have compared the effects of current and conductance noise in a neuronal model for subthreshold oscillations and action potential generation. We did not see any significant differences in the model behavior with respect to voltage traces, tuning curves of interspike intervals, interval distributions or frequency responses when the noise strength is adjusted. These findings indicate that simple current noise can give reasonable results in neuronal simulations with regard to physiological relevant noise effects.     

Small- and intermediate-conductance Ca2+-activated K+ channels directly control agonist-evoked nitric oxide synthesis in human vascular endothelial cells

The contribution of small-conductance (SK_Ca) and intermediate-conductance Ca²⁺-activated K⁺ (IK_Ca) channels to the generation of nitric oxide (NO) by Ca²⁺-mobilizing stimuli was investigated in human umbilical vein endothelial cells (HUVECs) by combining single-cell microfluorimetry with perforated patch-clamp recordings to monitor agonist-evoked NO synthesis, cytosolic Ca²⁺ transients, and membrane hyperpolarization in real time. ATP or histamine evoked reproducible elevations in NO synthesis and cytosolic Ca²⁺, as judged by 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) and fluo-3 fluorescence, respectively, that were tightly associated with membrane hyperpolarizations. Whereas evoked NO synthesis was unaffected by either tetraethylammonium (10 mmol/l) or BaCl₂ (50 µmol/l) + ouabain (100 µmol/l), depleting intracellular Ca²⁺ stores by thapsigargin or removing external Ca²⁺ inhibited NO production, as did exposure to high (80 mmol/l) external KCl. Importantly, apamin and charybdotoxin (ChTx)/ triarylmethane (TRAM)-34, selective blockers SK_Ca and IK_Ca channels, respectively, abolished both stimulated NO synthesis and membrane hyperpolarization and decreased evoked Ca²⁺ transients. Apamin and TRAM-34 also inhibited an agonist-induced outwardly rectifying current characteristic of SK_Ca and IK_Ca channels. Under voltage-clamp control, we further observed that the magnitude of agonist-induced NO production varied directly with the degree of membrane hyperpolarization. Mechanistically, our data indicate that SK_Ca and IK_Ca channel-mediated hyperpolarization represents a critical early event in agonist-evoked NO production by regulating the influx of Ca²⁺ responsible for endothelial NO synthase activation. Moreover, it appears that the primary role of agonist-induced release of intracellular Ca²⁺ stores is to trigger the opening of both K_Ca channels along with Ca²⁺ entry channels at the plasma membrane. Finally, the observed inhibition of stimulated NO synthesis by apamin and ChTx/TRAM-34 demonstrates that SK_Ca and IK_Ca channels are essential for NO-mediated vasorelaxation. calcium; endothelium; hyperpolarization; small-conductance calcium-activated potassium channel; intermediate-conductance calcium-activated potassium channel channel     

A "moving metal mechanism" for substrate cleavage by the DNA repair endonuclease APE-1

Apurinic/apyrimidinic endonuclease (APE-1) is essential for base excision repair (BER) of damaged DNA. Here molecular dynamics (MD) simulations of APE1 complexed with cleaved and uncleaved damaged DNA were used to determine the role and position of the metal ion(s) in the active site before and after DNA cleavage. The simulations started from an energy minimized wild-type structure of the metal-free APE1/damaged-DNA complex (1DE8). A grid search with one Mg2+ ion located two low energy clusters of Mg2+ consistent with the experimentally determined metal ion positions. At the start of the longer MD simulations, Mg2+ ions were placed at different positions as seen in the crystal structures and the movement of the ion was followed over the course of the trajectory. Our analysis suggests a "moving metal mechanism" in which one Mg2+ ion moves from the B- (more buried) to the A-site during substrate cleavage. The anticipated inversion of the phosphate oxygens occurs during the in-line cleavage reaction. Experimental results, which show competition between Ca2+ and Mg2+ for catalyzing the reaction, and high concentrations of Mg2+ are inhibitory, indicate that both sites cannot be simultaneously occupied for maximal activity.     

Hyaluronan is organized into fiber-like structures along migratory pathways in the developing mouse cerebellum.

Hyaluronan is a free glycosaminoglycan which is abundant in the extracellular matrix of the developing brain. Although not covalently linked to any protein it can act as a backbone molecule forming aggregates with chondroitin sulfate proteoglycans of the lectican family and link proteins. Using neurocan-GFP as a direct histochemical probe we analyzed the distribution and organization of hyaluronan in the developing mouse cerebellum, and related its fine structure to cell types of specified developmental stages. We observed a high affinity of this probe to fiber-like structures in the prospective white matter which are preferentially oriented parallel to the cerebellar cortex during postnatal development suggesting a specially organized form of hyaluronan. In other layers of the cerebellar cortex, the hyaluronan organization seemed to be more diffuse. During the second postnatal week, the overall staining intensity of hyaluronan in the white matter declined but fiber-like structures were still present at the adult stage. This type of hyaluronan organization is different from perineuronal nets e.g. found in deep cerebellar nuclei. Double staining experiments with cell type specific markers indicated that these fiber-like structures are predominantly situated in regions where motile cells such as Pax2-positive inhibitory interneuron precursors and MBP-positive oligodendroglial cells are located. In contrast, more stationary cells such as mature granule cells and Purkinje cells are associated with lower levels of hyaluronan in their environment. Thus, hyaluronan-rich fibers are concentrated at sites where specific neural precursor cell types migrate, and the anisotropic orientation of these fibers suggests that they may support guided neural migration during brain development.     

Vision: attending the invisible

In everyday vision, attention and awareness are hand in glove and almost impossible to tell apart. Recent work has exploited more contrived situations that allow these psychologically defined processes to be dissociated, providing insights into their respective neurophysiological correlates.     

Aggregate-centered redistribution of proteins by mutant huntingtin

Huntingtin is a widely expressed 350-kDa cytosolic multidomain of unknown function. Aberrant expansion of the polyglutamine tract located in the N-terminal region of huntingtin results in Huntington's disease. The presence of insoluble huntingtin inclusions in the brains of patients is one of the hallmarks of Huntington's disease. Experimentally, both full-length huntingtin and N-terminal fragments of huntingtin with expanded polyglutamine tracts trigger aggregate formation. Here, we report that upon the formation of huntingtin aggregates; endogenous cytosolic huntingtin, Hsc70/Hsp70 (heat shock protein and cognate protein of 70kDa) and syntaxin 1A become aggregate-centered. This redistribution suggests that these proteins are eventually depleted and become unavailable for normal cellular function. These results indicate that the cellular targeting of several key proteins are altered in the presence of mutant huntingtin and suggest that aggregate depletion of these proteins may underlie, in part, the sequence of disease progression.