Is bicarbonate in Photosystem II the equivalent of the glutamate ligand to the iron atom in bacterial reaction centers?

Photosystem II of oxygen-evolving organisms exhibits a bicarbonate-reversible formate effect on electron transfer between the primary and secondary acceptor quinones, QA and QB. This effect is absent in the otherwise similar electron acceptor complex of purple bacteria, e.g. Rhodobacter sphaeroides. This distinction has led to the suggestion that the iron atom of the acceptor quinone complex in PS II might lack the fifth and sixth ligands provided in the bacterial reaction center (RC) by a glutamate residue at position 234 of the M-subunit in Rb. sphaeroides,RCs (M232 in Rps. viridis). By site-directed mutagenesis we have altered GluM234 in RCs from Rb. sphaeroides, replacing it with valine, glutamine and glycine to form mutants M234EV, M234EQ and M234EG, respectively. These mutants grew competently under phototrophic conditions and were tested for the formate-bicarbonate effect. In chromatophores there were no detectable differences between wild type (Wt) and mutant M234EV with respect to cytochrome b-561 reduction following a flash, and no effect of bicarbonate depletion (by incubation with formate). In isolated RCs, several electron transfer activities were essentially unchanged in Wt and M234EV, M234EQ and M234EG mutants, and no formate-bicarbonate effect was observed on: (a) the fast or slow phases of recovery of the oxidized primary donor (P+) in the absence of exogenous donor, i.e., the recombination of P+QA− or P+QB−, respectively; (b) the kinetics of electron transfer from QA− to QB; or (c) the flash dependent oscillations of semiquinone formation in the presence of donor to P+ (QB turnover). The absence of a formate-bicarbonate effect in these mutants suggests that GluM234 is not responsible for the absence of the formate-bicarbonate effect in Wt bacterial RCs, or at least that other factors must be taken into account. The mutant RCs were also examined for the fast primary electron transfer along the active (A-)branch of the pigment chain, leading to reduction of QA. The kinetics were resolved to reveal the reduction of the monomer bacteriochlorophyll (τ = 3.5 ps), followed by reduction of the bacteriopheophytin (τ = 0.9 ps). Both steps were essentially unaltered from the wild type. However, the rate of reduction of QA was slowed by a factor of 2 (τ = 410 ± 30 and 47 ± 30 ps for M234EQ and M234EV, respectively, compared to 220 ps in the wild type). EPR studies of the isolated RCs showed a characteristic g = 1.82 signal for the QA semiquinone coupled to the iron atom, which was indistinguishable from the wild type. It is concluded that GluM234 is not essential to the normal functioning of the acceptor quinone complex in bacterial RCs and that the role of bicarbonate in PS II is distinct from the role of this residue in bacterial RCs

Modeling of Spiking-Bursting Neural Behavior Using Two-Dimensional Map

A simple model that replicates the dynamics of spiking and spiking-bursting activity of real biological neurons is proposed. The model is a two-dimensional map which contains one fast and one slow variable. The mechanisms behind generation of spikes, bursts of spikes, and restructuring of the map behavior are explained using phase portrait analysis. The dynamics of two coupled maps which model the behavior of two electrically coupled neurons is discussed. Synchronization regimes for spiking and bursting activity of these maps are studied as a function of coupling strength. It is demonstrated that the results of this model are in agreement with the synchronization of chaotic spiking-bursting behavior experimentally found in real biological neurons.Comment: 9 pages, 12 figure

Stable Propagation of a Burst Through a One-Dimensional Homogeneous Excitatory Chain Model of Songbird Nucleus HVC

We demonstrate numerically that a brief burst consisting of two to six spikes can propagate in a stable manner through a one-dimensional homogeneous feedforward chain of non-bursting neurons with excitatory synaptic connections. Our results are obtained for two kinds of neuronal models, leaky integrate-and-fire (LIF) neurons and Hodgkin-Huxley (HH) neurons with five conductances. Over a range of parameters such as the maximum synaptic conductance, both kinds of chains are found to have multiple attractors of propagating bursts, with each attractor being distinguished by the number of spikes and total duration of the propagating burst. These results make plausible the hypothesis that sparse precisely-timed sequential bursts observed in projection neurons of nucleus HVC of a singing zebra finch are intrinsic and causally related.Comment: 13 pages, 6 figure

Cell sorting in a Petri dish controlled by computer vision.

Fluorescence-activated cell sorting (FACS) applying flow cytometry to separate cells on a molecular basis is a widespread method. We demonstrate that both fluorescent and unlabeled live cells in a Petri dish observed with a microscope can be automatically recognized by computer vision and picked up by a computer-controlled micropipette. This method can be routinely applied as a FACS down to the single cell level with a very high selectivity. Sorting resolution, i.e., the minimum distance between two cells from which one could be selectively removed was 50-70 micrometers. Survival rate with a low number of 3T3 mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was 66 +/- 12% and 88 +/- 16%, respectively. Purity of sorted cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures were 95 +/- 2% and 62 +/- 7%, respectively. Hydrodynamic simulations confirmed the experimental sorting efficiency and a cell damage risk similar to that of normal FACS

Tactile Discrimination Using Template Classifiers: Towards a Model of Feature Extraction in Mammalian Vibrissal Systems

Rats and other whiskered mammals are capable of making sophisticated sensory discriminations using tactile signals from their facial whiskers (vibrissae). As part of a programme of work to develop biomimetic technologies for vibrissal sensing, including whiskered robots, we are devising algorithms for the fast extraction of object parameters from whisker deflection data. Previous work has demonstrated that radial distance to contact can be estimated from forces measured at the base of the whisker shaft. We show that in the case of a moving object contacting a whisker, the measured force can be ambiguous in distinguishing a nearby object moving slowly from a more distant object moving rapidly. This ambiguity can be resolved by simultaneously extracting object position and speed from the whisker deflection time series – that is by attending to the dynamics of the whisker’s interaction with the object. We compare a simple classifier with an adaptive EM (Expectation Maximisation) classifier. Both systems are effective at simultaneously extracting the two parameters, the EM-classifier showing similar performance to a handpicked template classifier. We propose that adaptive classification algorithms can provide insights into the types of computations performed in the rat vibrissal system when the animal is faced with a discrimination task

Statistical mechanics of temporal association in neural networks with transmission delays

We study the representation of static patterns and temporal sequences in neural networks with signal delays and a stochastic parallel dynamics. For a wide class of delay distributions, the asymptotic network behavior can be described by a generalized Gibbs distribution, generated by a novel Lyapunov functional for the determination dynamics. We extend techniques of equilibrium statistical mechanics so as to deal with time-dependent phenomena, derive analytic results for both retrieval quality and storage capacity, and compare them with numerical simulations

Dynamical model of sequential spatial memory: winnerless competition of patterns

We introduce a new biologically-motivated model of sequential spatial memory which is based on the principle of winnerless competition (WLC). We implement this mechanism in a two-layer neural network structure and present the learning dynamics which leads to the formation of a WLC network. After learning, the system is capable of associative retrieval of pre-recorded sequences of spatial patterns.Comment: 4 pages, submitted to PR

Identification of Neural Circuits by Imaging Coherent Electrical Activity with FRET-Based Dyes

AbstractWe show that neurons that underlie rhythmic patterns of electrical output may be identified by optical imaging and frequency-domain analysis. Our contrast agent is a two-component dye system in which changes in membrane potential modulate the relative emission between a pair of fluorophores. We demonstrate our methods with the circuit responsible for fictive swimming in the isolated leech nerve cord. The output of a motor neuron provides a reference signal for the phase-sensitive detection of changes in fluorescence from individual neurons in a ganglion. We identify known and possibly novel neurons that participate in the swim rhythm and determine their phases within a cycle. A variant of this approach is used to identify the postsynaptic followers of intracellularly stimulated neurons

Lasing without inversion in three-level systems without external coherent driving

We have studied an incoherently pumped laser operating with a Doppler-broadened three-level system placed in a doubly resonant cavity. This system generates two laser fields, one of them without population inversion. Both ladder and V-type three-level schemes are considered with a ratio R=ωα/ωβ of inversionless laser frequency ωα to ordinary laser frequency ωβ of R=0.67 and R=1.88, respectively. Dual-wavelength lasing extends up to Doppler-broadening values for optical transitions of atoms in a vapor cell. Some considerations for the practical realization of this dual-wavelength laser are discussed