Gait switching with phase reversal of locomotory waves in the centipedeScolopocryptops rubiginosus.

Crawling using locomotory waves is a common method of locomotion for limbless and many-legged invertebrates and stimulates the biomimetic engineering of flexible locomotion. It is generally believed that the direction of locomotory waves is fixed for a given species. However, we found that a centipede,Scolopocryptops rubiginosus, flexibly generated its gait to allow for locomotory waves that varied in direction, depending on (i) locomotion speed and (ii) the physical conditions of terrain. We also found a new type of centipede's swimming gait unlike eel-like way known so far which is using posteriorly traveling waves of horizontal body undulation. The gait patterns of the centipede were examined in various conditions and analyzed how the waves switched in detailed. We showed that gait patterns were associated with control of stride length rather than stride frequency. Discussion was made on a possible scenario of the gait transition in the centipede compatible with our observations. This finding may give a hint at bio-inspired control of flexible gait switching in response to irregular terrain.

Physiological properties of Cantor coding-like iterated function system in the hippocampal CA1 network.

Cantor coding provides an information coding scheme for temporal sequences of events. In the hippocampal CA3-CA1 network, Cantor coding-like mechanism was observed in pyramidal neurons and the relationship between input pattern and recorded responses could be described as an iterated function system. However, detailed physiological properties of the system in CA1 remain unclear. Here, we performed a detailed analysis of the properties of the system related to the physiological basis of learning and memory. First, we investigated whether the system could be simply based on a series of on-off responses of excitatory postsynaptic potential (EPSP) amplitudes. We applied a series of three spatially distinct input patterns with similar EPSP peak amplitudes. The membrane responses showed significant differences in spatial clustering properties related to the iterated function system. These results suggest that existence of some factors, which do not simply depend on a series of on-off responses but on spatial patterns in the system. Second, to confirm whether the system is dependent on the interval of sequential input, we applied spatiotemporal sequential inputs at several intervals. The optimal interval was 30 ms, similar to the physiological input from CA3 to CA1. Third, we analyzed the inhibitory network dependency of the system. After GABAA receptor blocker (gabazine) application, quality of code discrimination in the system was lower under subthreshold conditions and higher under suprathreshold conditions. These results suggest that the inhibitory network increase the difference between the responses under sub- and suprathreshold conditions. In summary, Cantor coding-like iterated function system appears to be suitable for information expression in relation to learning and memory in CA1 network.

Localization of photoperiod responsive circadian oscillators in the mouse suprachiasmatic nucleus.

The circadian pacemaker in the suprachiasmatic nucleus (SCN) yields photoperiodic response to transfer seasonal information to physiology and behavior. To identify the precise location involved in photoperiodic response in the SCN, we analyzed circadian Period1 and PERIOD2 rhythms in horizontally sectioned SCN of mice exposed to a long or short day. Statistical analyses of bioluminescence images with respective luciferase reporters on pixel level enabled us to identify the distinct localization of three oscillating regions; a large open-ring-shape region, the region at the posterior end and a sharply demarcated oval region at the center of the SCN. The first two regions are the respective sites for the so-called evening and morning oscillators, and the third region is possibly a site for mediating photic signals to the former oscillators. In these regions, there are two classes of oscillating cells in which Per1 and Per2 could play differential roles in photoperiodic responses.

Current reinforcement model reproduces center-in-center vein trajectory of Physarum polycephalum.

Vein networks span the whole body of the amoeboid organism in the plasmodial slime mould Physarum polycephalum, and the network topology is rearranged within an hour in response to spatio-temporal variations of the environment. It has been reported that this tube morphogenesis is capable of solving mazes, and a mathematical model, named the 'current reinforcement rule', was proposed based on the adaptability of the veins. Although it is known that this model works well for reproducing some key characters of the organism's maze-solving behaviour, one important issue is still open: In the real organism, the thick veins tend to trace the shortest possible route by cutting the corners at the turn of corridors, following a center-in-center trajectory, but it has not yet been examined whether this feature also appears in the mathematical model, using corridors of finite width. In this report, we confirm that the mathematical model reproduces the center-in-center trajectory of veins around corners observed in the maze-solving experiment.

Dissociation of Per1 and Bmal1 circadian rhythms in the suprachiasmatic nucleus in parallel with behavioral outputs.

The temporal order of physiology and behavior in mammals is primarily regulated by the circadian pacemaker located in the hypothalamic suprachiasmatic nucleus (SCN). Taking advantage of bioluminescence reporters, we monitored the circadian rhythms of the expression of clock genes Per1 and Bmal1 in the SCN of freely moving mice and found that the rate of phase shifts induced by a single light pulse was different in the two rhythms. The Per1-luc rhythm was phase-delayed instantaneously by the light presented at the subjective evening in parallel with the activity onset of behavioral rhythm, whereas the Bmal1-ELuc rhythm was phase-delayed gradually, similar to the activity offset. The dissociation was confirmed in cultured SCN slices of mice carrying both Per1-luc and Bmal1-ELuc reporters. The two rhythms in a single SCN slice showed significantly different periods in a long-term (3 wk) culture and were internally desynchronized. Regional specificity in the SCN was not detected for the period of Per1-luc and Bmal1-ELuc rhythms. Furthermore, neither is synchronized with circadian intracellular Ca2+ rhythms monitored by a calcium indicator, GCaMP6s, or with firing rhythms monitored on a multielectrode array dish, although the coupling between the circadian firing and Ca2+ rhythms persisted during culture. These findings indicate that the expressions of two key clock genes, Per1 and Bmal1, in the SCN are regulated in such a way that they may adopt different phases and free-running periods relative to each other and are respectively associated with the expression of activity onset and offset.

Dual origins of the intracellular circadian calcium rhythm in the suprachiasmatic nucleus.

In mammals, the master circadian clock is located in the suprachiasmatic nucleus (SCN), where most neurons show circadian rhythms of intracellular Ca2+ levels. However, the origin of these Ca2+ rhythms remains largely unknown. In this study, we successfully monitored the intracellular circadian Ca2+ rhythms together with the circadian PER2 and firing rhythms in a single SCN slice ex vivo, which enabled us to explore the origins. The phase relation between the circadian PER2 and Ca2+ rhythms, but not between the circadian PER2 and firing rhythms, was significantly altered in Cry1/Cry2 double knockout mice, which display a loss of intercellular synchronization in the SCN. In addition, in Cry1/Cry2 double knockout mice, circadian Ca2+ rhythms were abolished in the dorsolateral SCN, but were maintained in the majority of the ventromedial SCN. These findings indicate that intracellular circadian Ca2+ rhythms are composed of an exogenous and endogenous component involving PER2 expression.

A ciliate memorizes the geometry of a swimming arena.

Previous studies on adaptive behaviour in single-celled organisms have given hints to the origin of their memorizing capacity. Here we report evidence that a protozoan ciliate Tetrahymena has the capacity to learn the shape and size of its swimming space. Cells confined in a small water droplet for a short period were found to recapitulate circular swimming trajectories upon release. The diameter of the circular trajectories and their duration reflected the size of the droplet and the period of confinement. We suggest a possible mechanism for this adaptive behaviour based on a Ca(2+) channel. In our model, repeated collisions with the walls of a confining droplet result in a slow rise in intracellular calcium that leads to a long-term increase in the reversal frequency of the ciliary beat.

Allometry in Physarum plasmodium during free locomotion: size versus shape, speed and rhythm.

Physarum plasmodium is a giant unicellular organism whose length can vary by more than three orders of magnitude. Using plasmodia ranging in size from 100 µm to 10 cm, we investigated the size dependency of their thickness distributions and locomotion speeds during free locomotion. (1) In the longitudinal direction, the organism is thickest close to the front, and decreases exponentially in thickness towards the rear. The slenderness ratio varies with body size according to a power law, such that large plasmodia are long and flat, whereas small plasmodia are short and thick. (2) The mean locomotion speed is proportional to the mean maximum thickness of the frontal part. By conducting a dimensional analysis, possible physical models are discussed. (3) The intrinsic period of the thickness oscillation, which is related to shuttle streaming (period 1-2 min), increases logarithmically with body size. (4) Various characteristics exhibit size-independent, long-period (20±10 min) oscillations, including speed, shape and intrinsic thickness oscillation period. These variations are closely coupled to formation of the entire cell shape, including undulation of thickness along the longitudinal axis and timing of branching of the frontal tip. Based on these experimental results and those reported previously, we propose a simple mathematical model for cell locomotion.

Attempts to retreat from a dead-ended long capillary by backward swimming in Paramecium.

We have observed how the ciliate Paramecium attempts to retreat from the dead-end of a long capillary that is too narrow for turning. After many trial-and-error episodes of short-term backward swimming (SBS), which is the conventional avoidance behavior exhibited in free swimming when an obstacle is faced, long-term backward swimming (LBS) that lasted five to ten times longer was developed. LBS may have a beneficial effect for complete withdrawal from the capillary space, although in our experiment it was impossible for the organism to do so due to the capillary length. In order to identify a physically possible mechanism for LBS, we propose model equations for the membrane potential of Hodgkin-Huxley type, which describe the control of ciliary movement. The physiological implications and physical mechanism of the development of LBS are discussed.

Common mechanics of mode switching in locomotion of limbless and legged animals.

Crawling using muscular waves is observed in many species, including planaria, leeches, nemertea, aplysia, snails, chitons, earthworms and maggots. Contraction or extension waves propagate along the antero-posterior axis of the body as the crawler pushes the ground substratum backward. However, the observation that locomotory waves can be directed forward or backward has attracted much attention over the past hundred years. Legged organisms such as centipedes and millipedes exhibit parallel phenomena; leg tips form density waves that propagate backward or forward. Mechanical considerations reveal that leg-density waves play a similar role to locomotory waves in limbless species, and that locomotory waves are used by a mechanism common to both legged and limbless species to achieve crawling. Here, we report that both mode switching of the wave direction and friction control were achieved when backward motion was induced in the laboratory. We show that the many variations of switching in different animals can essentially be classified in two types according to mechanical considerations. We propose that during their evolution, limbless crawlers first moved in a manner similar to walking before legs were obtained. Therefore, legged crawlers might have learned the mechanical mode of movement involved in walking long before obtaining legs.

Topological specificity and hierarchical network of the circadian calcium rhythm in the suprachiasmatic nucleus.

The circadian pacemaker in the hypothalamic suprachiasmatic nucleus (SCN) is a hierarchical multioscillator system in which neuronal networks play crucial roles in expressing coherent rhythms in physiology and behavior. However, our understanding of the neuronal network is still incomplete. Intracellular calcium mediates the input signals, such as phase-resetting stimuli, to the core molecular loop involving clock genes for circadian rhythm generation and the output signals from the loop to various cellular functions, including changes in neurotransmitter release. Using a unique large-scale calcium imaging method with genetically encoded calcium sensors, we visualized intracellular calcium from the entire surface of SCN slice in culture including the regions where autonomous clock gene expression was undetectable. We found circadian calcium rhythms at a single-cell level in the SCN, which were topologically specific with a larger amplitude and more delayed phase in the ventral region than the dorsal. The robustness of the rhythm was reduced but persisted even after blocking the neuronal firing with tetrodotoxin (TTX). Notably, TTX dissociated the circadian calcium rhythms between the dorsal and ventral SCN. In contrast, a blocker of gap junctions, carbenoxolone, had only a minor effect on the calcium rhythms at both the single-cell and network levels. These results reveal the topological specificity of the circadian calcium rhythm in the SCN and the presence of coupled regional pacemakers in the dorsal and ventral regions. Neuronal firings are not necessary for the persistence of the calcium rhythms but indispensable for the hierarchical organization of rhythmicity in the SCN.

A mathematical model for Cantor coding in the hippocampus.

Recent studies suggest that the hippocampus is crucial for memory of sequentially organized information. Cantor coding in hippocampal CA1 is theoretically hypothesized to provide a scheme for encoding temporal sequences of events. Here, in order to investigate this Cantor coding in detail, we construct a CA1 network model consisting of conductance-based model neurons. It is assumed that CA3 outputs temporal sequences of spatial patterns to CA1. We examine the dependence of output patterns of CA1 neurons on input time series by taking each output and combining it with an input sequence. It is shown that the output patterns of CA1 were hierarchically clustered in a self-similar manner according to the similarity of input temporal sequences. The population dynamics of the network can be well approximated by a set of contractive affine transformations, which forms a Cantor set. Furthermore, it is shown that the performance of the encoding scheme sensitively depends on the interval of input sequences. The bursting neurons with NMDA synapses are effective for encoding sequential input with long (over 150 ms) intervals while the non-bursting neurons with AMPA synapses are effective for encoding input with short (less than 30 ms) intervals.

Iterated function systems in the hippocampal CA1.

How does the information of spatiotemporal sequence stemming from the hippocampal CA3 area affect the postsynaptic membrane potentials of the hippocampal CA1 neurons? In a recent study, we observed hierarchical clusters of the distribution of membrane potentials of CA1 neurons, arranged according to the history of input sequences (Fukushima et al Cogn Neurodyn 1(4):305-316, 2007). In the present paper, we deal with the dynamical mechanism generating such a hierarchical distribution. The recording data were investigated using return map analysis. We also deal with a collective behavior at population level, using a reconstructed multi-cell recording data set. At both individual cell and population levels, a return map of the response sequence of CA1 pyramidal cells was well approximated by a set of contractive affine transformations, where the transformations represent self-organized rules by which the input pattern sequences are encoded. These findings provide direct evidence that the information of temporal sequences generated in CA3 can be self-similarly represented in the membrane potentials of CA1 pyramidal cells.

Introduction of the disulfide proteome: application of a technique for the analysis of plant storage proteins as well as allergens.

Redox regulation plays an important role across a broad spectrum of biology. Accumulating evidence suggests that thioredoxin, a widely distributed redox enzyme, participates in the redox control of numerous target proteins, thus, playing a key role as a signaling intermediate that senses the metabolic state or environmental changes, and transmits information to induce appropriate reactions for survival. The recent development of proteomic strategies has facilitated the identification of thioredoxin targets to allow clarification of new thioredoxin-dependent redox mechanisms. This review examines the usefulness of a disulfide proteome technique for the analysis of thioredoxin-dependent redox regulation as well as for use in allergen studies.

Spatial clustering property and its self-similarity in membrane potentials of hippocampal CA1 pyramidal neurons for a spatio-temporal input sequence.

To clarify how the information of spatiotemporal sequence of the hippocampal CA3 affects the postsynaptic membrane potentials of single pyramidal cells in the hippocampal CA1, the spatio-temporal stimuli was delivered to Schaffer collaterals of the CA3 through a pair of electrodes and the post-synaptic membrane potentials were recorded using the patch-clamp recording method. The input-output relations were sequentially analyzed by applying two measures; "spatial clustering" and its "self-similarity" index. The membrane potentials were hierarchically clustered in a self-similar manner to the input sequences. The property was significantly observed at two and three time-history steps. In addition, the properties were maintained under two different stimulus conditions, weak and strong current stimulation. The experimental results are discussed in relation to theoretical results of Cantor coding, reported by Tsuda (Behav Brain Sci 24(5):793-847, 2001) and Tsuda and Kuroda (Jpn J Indust Appl Math 18:249-258, 2001; Cortical dynamics, pp 129-139, Springer-Verlag, 2004).

Disulfide proteome in the analysis of protein function and structure.

Many proteins undergo post-translational modification via well defined mechanisms such as acetylation, phosphorylation and glycosylation and thereby control a spectrum of biochemical processes. A growing body of evidence suggests that the reversible reduction of disulfide bonds also alters the structure and activity of proteins. Thioredoxin, a ubiquitous 12 kDa protein with a catalytically active disulfide active site (Cys-Gly-Pro-Cys), plays a central role in controlling the redox status of disulfide bonds in proteins that regulate a range of processes. Included are photosynthesis, seed germination, transcription, cell division, radical scavenging and detoxification. The ability to identify unknown functions of proteins of all types has been advanced by the emerging field of functional proteomics. In this brief review, we introduce the disulfide proteome as a tool that complements other methods for the comprehensive analysis of proteins. In so doing, the usefulness of applying this method for both in vitro and in vivo analyses is discussed for thioredoxin and other disulfide proteins, especially those occurring in plants.