Synchronization of mechanochemical auto-oscillations within the Physarum polycephalum plasmodium by periodical external actions

Amoeboid locomotion of huge unicellular organism, the Physarum polycephalum plasmodium, is stipulated by endoplasmic flow, which is produced by spatially highly coordinated rhythmic contractions of the ectoplasm. To describe the self-organization of the plasmodial contractile activity, we proposed a mathematical model, which is based on the hypothesis of positive feedback between the deformation of the cytoskeleton and release of a chemical regulator of the active contraction. A nonautonomous analogue of this model was used to study the synchronization of mechanochemical auto-oscillations by periodic gradient of the external pressure. Numerical computations of the system of differential equations obtained revealed a dependency of the synchronization band on the amplitude of the external pressure oscillations. On the basis of this dependence and experimental data on the band of synchronization of the shuttle endoplasmic flow by the periodic gradient of temperature obtained with the help of the laser Doppler anemometer, relative efficiency of external synchronizing action of temperature and pressure was evaluated.     

Synchronization of wing beat cycle of the desert locust, Schistocerca gregaria, by periodic light flashes

Studies on the generation of rhythmic motor patterns have shown that peripheral sensory input may contribute substantially to the rhythm generating network. A prominent example is the wing beat frequency of desert locusts, which can be entrained to rhythmic mechanosensory input, but also to the frequency of periodic light flashes. To further analyze the entrainment by light, tethered flying locusts were presented with periodic light flashes, while the position of the forewing was filmed. We show that entrainment of wing beat occurs both in the UV and green range of light. Animals maintained a characteristic phase relationship to the light stimulus with the most elevated wing position occurring at the end of the dark phase. Speed and time course of entrainment varied greatly and ranged from the duration of a single wing beat cycle to several seconds. To identify the visual system mediating entrainment, synchronization to UV light was tested after cutting the optic stalks to the optic lobes/compound eyes or the ocellar nerves. The results show that light entrainment of the locust flight pattern is largely and perhaps exclusively mediated via the fast ocellar pathway and may have a role to stabilize flight with respect to the horizon.     

Calcium wave for cytoplasmic streaming of Physarum polycephalum

The plasmodium Physarum polycepharum exhibits periodic cycles of cytoplasmic streaming in association with those of contraction and relaxation movement. In the present study, we injected Calcium Green dextran as a fluorescent Ca²⁺ indicator into the thin‐spread living plasmodium. We found changes in the [Ca²⁺]i (intracellular concentration of Ca²⁺), which propagated in a wave‐like form in its cytoplasm. The Ca²⁺ waves were also detected when we used Fura dextran which detected [Ca²⁺]i by the ratio of two wavelengths. We prepared the plasmodial fragment from the thin‐spread and found that the cycles of the contraction–relaxation movement was so synchronized that the measurement of its area provided an indication of the movement. We observed that [Ca²⁺]i also synchronized in the entire fragment and that the relaxation ensued upon the reduction in [Ca²⁺]i. We suggest that the Ca²⁺ wave generated periodically is one of the major factors playing a crucial role in the relaxation of P. polycepharum.     

Experimental study of synchronization phenomena under periodic light irradiation of a nonlinear chemical system

A photosensitive chemical oscillating reaction, i.e., the Briggs-Rauscher (B.R.) reaction, exhibiting a wealth of nonlinear behavior, when performed in a continuous-flow stirred-tank reactor, and subjected to periodic light irradiation, is studied as an experimental example of entrainment phenomena observable in biological systems. The adaptation patterns under periodic light irradiation are elucidated by means of the response of the system to continuous and single-pulse light irradiation. It is shown that self-oscillating states, excitable steady states and bistable systems can exhibit the same types of synchronization patterns when submitted to periodic external forces with appropriate amplitude and time scale conditions.     

Patterns of oscillation during mitosis in plasmodia ofPhysarum polycephalum

Summary The rhythmic contraction pattern in plasmodia ofPhysarum polycephalum was studied to determine whether characteristic changes occur during the synchronized nuclear division. An electrical method that measures the contraction rhythm in situ during several cell cycles was used. Biopsies of the plasmodia were taken at 17 min intervals for precise determination of the cell cycle stages and were correlated with the simultaneously measured contraction rhythm. All measurements were performed in a temperature controlled environment (27 °C) at 100% relative humidity with the plasmodia (less than 24 h old) growing on a semi-defined agar medium. A total of 14 different plasmodia have been examined, and on one occasion the plasmodium was followed through 3 subsequent mitoses. The mitotic stages were identified with aceto-orcein coloring techniques and by fluorescence methods. Except for a few cases where a mitotic asynchrony of 2–3 min was observed, the mitotic events occurred simultaneously in the nuclei within a single plasmodium. Both the occurrence of the first mitosis after inoculation and the intermitotic times were highly variable. Our study indicates that the contraction rhythm in plasmodia ofPhysarum is unperturbed during the synchronized nuclear division. However, in 5 of the 17 examined mitoses an amplitude decay was observed. We discuss possible explanations for the obtained results with emphasis on the applied techniques, interpretation of the oscillation patterns, and possible restrictions in the cell itself.     

Synthesis of mRNA is Required for Light-Induced Phase Shifting of the Circadian Conidiation Rhythm in Neurospora crassa

The process of light-induced phase shifting was investigatedin Neurospora crassa using a liquid culture system and a combinationof treatment with a nucleoside analogue and light. 5-Azacytidineinhibited the light-induced phase shifting at all phases thatwere sensitive to light. Electrophoresis of proteins that weresynthesized in a translation system in vitro showed that 5-azacytidineinhibited the synthesis of most mRNAs. The inhibition of mRNAsynthesis was correlated with the inhibition of light-inducedphase shifting. An excess of cytidine completely overcame theinhibition by 5-azacytidine of both light-induced phase shiftingand mRNA synthesis. Other analogues, namely, 6-azauridine and6-methylpurine, failed to inhibit either the light-induced phaseshifting or the synthesis of mRNA. Two-dimensional gel electrophoresisshowed that the levels of expression of nine mRNAs were affectedby light within 30 min after irradiation. By contrast, the oscillatorof the circadian clock was not affected by pulse treatment with5-azacytidine alone because such treatment failed to shift thephase of the circadian rhythm at any phase. These results indicatethat newly synthesized mRNA(s) is required during the processof signal transduction, from the light-perceiving system tothe circadian clock, for light-induced phase shifting in Neurospora. (Received October 17, 1994; Accepted January 23, 1995)     

Interaction between cell shape and contraction pattern in the Physarum plasmodium

The relationship between cell shape and rhythmic contractile activity in the large amoeboid organism Physarum polycephalum was studied. The organism develops intricate networks of veins in which protoplasmic sol moved to and fro very regularly. When migrating on plain agar, the plasmodium extends like a sheet and develops dendritic veins toward the rear. After a particular stimulation, the vein organization changes into veinless or vein-network structures. In both structures, the mixing rate of the protoplasm, which is related to communication among contraction oscillators, decreased compared with that of the dendritic one. Accompanying these changes in vein structure, the spatio-temporal pattern of the rhythmic contraction changed into a small-structured pattern from a synchronized one. In the above process, cell shape affects the contraction pattern, but, conversely, the contraction pattern effects the cell shape. To demonstrate this, a phase difference in the rhythmic contraction was induced artificially by entraining the intrinsic rhythm to external temperature oscillations. New veins then formed along the direction parallel to the phase difference of the rhythm. Consequently, the vein organization of the cell interacts with the contractile activity to form a feedback loop in a mechanism of contraction pattern formation.     

Cardiorespiratory synchronism: revealing in human,dependence on properties of nervous system and functional states of an organism

Cardiorespiratory synchronism (CRS) shows that at the breathing frequency, which as usual exceeds the baseline heart rhythm, the heart makes one contraction per each breathing. It is shown that CRS arises as a result of reproduction by the heart the rhythm of signals, coming to the heart via vagus nerves. CRS characterized by the synchronization range, duration of its development after the beginning of the rapid respiration, difference between the baseline heartbeat frequency and lower limit of synchronization range. The CRS parameters were determined in humans at the age of 5-65. The CRS parameters depends on nervous system type, vegetative tonus of the nervous system and functional states of an organism.     

LASER MICROSCOPE IRRADIATION OF PHYSARUM POLYCEPHALUM: DYNAMIC AND ULTRASTRUCTURAL EFFECTS

Streaming plasmodia of Physarum polycephalum were irradiated with a microscope-mounted ruby laser and the resulting changes were recorded by cinemicrography or streak photographs. Some lesions were processed for electron microscopy. By varying the incident energy, three levels of response were detected. Two transient responses, a gelation briefly blocking streams and a more severe gelation with contraction, changed movement patterns but not organelle ultrastructure. At higher energies, a permanently coagulated lesion was rapidly segregated from normal and transiently altered cytoplasm by formation of new membranes. Within the coagulum, pigment granules were destroyed, membranes were disrupted, and cytoplasm was flocculent. Nuclei and mitochondria were compact in the center and swollen in a peripheral space left by contraction of the coagulum. These changes are probably caused by heat produced by the interaction between the laser beam and the pigment granules of the plasmodium. Many of the changes seem to be secondary responses that follow the primary capture of energy during irradiation.     

Changes in the fluctuation of interbeat intervals in spontaneously beating cultured cardiac myocytes: experimental and modeling studies

 Isolated and cultured neonatal cardiac myocytes contract spontaneously and cyclically, and have the properties of a non-linear oscillator. In this study, we have analyzed the relationship between the fluctuation of contraction rhythm of spontaneously beating cultured cardiac myocytes, and the coupling strength among them. The coefficient of variation of contraction intervals increased transiently in the early stages of incubation, and then decreased almost monotonically with time. The contraction rhythm of the myocytes became synchronized in the late stage of the culture. The day on which synchronization occurred almost coincided with the day when the coefficient of variation reached its lowest value. In addition, we have performed a mathematical analysis using interacting Bonhoeffer–van der Pol oscillators to clarify the mechanisms underlying the changes in the fluctuation of contraction rhythm with time. As the coupling strength among oscillators increased, the coefficient of variation of oscillation periods increased temporarily, but then decreased rapidly when the oscillators showed synchronization. These results suggest that the changes in the fluctuation of beating rhythm result from the increase in strength of electrical coupling among spontaneously beating cardiac myocytes. Received: 10 August 2000 / Accepted in revised form: 19 August 2001     

The avoidance and aggregative movements of mesophyll chloroplasts in C(4) monocots in response to blue light and abscisic acid

In C(4) plants, mesophyll (M) chloroplasts are randomly distributed along the cell walls, whereas bundle sheath chloroplasts are located in either a centripetal or centrifugal position. It was reported previously that only M chloroplasts aggregatively redistribute to the bundle sheath side in response to extremely strong light or environmental stresses. The aggregative movement of M chloroplasts is also induced in a light-dependent fashion upon incubation with abscisic acid (ABA). The involvement of reactive oxygen species (ROS) and red/blue light in the aggregative movement of M chloroplasts are examined here in two distinct subtypes of C(4) plants, finger millet and maize. Exogenously applied hydrogen peroxide or ROS scavengers could not change the response patterns of M chloroplast movement to light and ABA. Blue light irradiation essentially induced the rearrangement of M chloroplasts along the sides of anticlinal walls, parallel to the direction of the incident light, which is analogous to the avoidance movement of C(3) chloroplasts. In the presence of ABA, most of the M chloroplasts showed the aggregative movement in response to blue light but not red light. Together these results suggest that ROS are not involved in signal transduction for the aggregative movement, and ABA can shift the blue light-induced avoidance movement of C(4)-M chloroplasts to the aggregative movement.     

Circadian regulation of retinomotor movements. I. Interaction of melatonin and dopamine in the control of cone length

In lower vertebrates, cone retinomotor movements occur in response to changes in lighting conditions and to an endogenous circadian clock. In the light, cone myoids contract, while in the dark, they elongate. In order to test the hypothesis that melatonin and dopamine may be involved in the regulation of cone movement, we have used an in vitro eyecup preparation from Xenopus laevis that sustains light- and dark-adaptive cone retinomotor movement. Melatonin mimics darkness by causing cone elongation. Dark- and melatonin-induced cone elongation are blocked by dopamine. Dopamine also stimulates cone contraction in dark-adapted eyecups. The effect of dopamine appears to be mediated specifically by a dopamine receptor, possibly of the D2 type. The dopamine agonist apomorphine and the putative D2 agonist LY171555 induced cone contraction. In contrast, the putative D1 agonist SKF38393-A and specific alpha 1-, alpha 2-, and beta-adrenergic receptor agonists were without effect. Furthermore, the dopamine antagonist spiroperidol not only blocked light-induced cone contraction, but also stimulated cone elongation in the light. These results suggest that dopamine is part of the light signal for cone contraction, and that its suppression is part of the dark signal for cone elongation. Melatonin may affect cone movement indirectly through its influence on the dopaminergic system.     

TheBulla ocular circadian pacemaker

In an effort to understand the cellular basis of entrainment of circadian oscillators we have studied the role of membrane potential changes in the neurons which comprise the ocular circadian pacemaker of Bulla gouldiana in mediating phase shifts of the ocular circadian rhythm. We report that: 1. Intracellular recording was used to measure directly the effects of the phase shifting agents light, serotonin, and 8-bromo-cAMP on the membrane potential of the basal retinal neurons. We found that light pulses evoke a transient depolarization followed by a smaller sustained depolarization. Application of serotonin produced a biphasic response; a transient depolarization followed by a sustained hyperpolarization. Application of a membrane permeable analog of the intracellular second messenger cAMP, 8-bromo-cAMP, elicited sustained hyperpolarization, and occasionally a weak phasic depolarization. 2. Changing the membrane potential of the basal retinal neurons directly and selectively with intracellularly injected current phase shifts the ocular circadian rhythm. Both depolarizing and hyperpolarizing current can shift the phase of the circadian oscillator. Depolarizing current mimics the phase shifting action of light, while hyperpolarizing current produces phase shifts which are transposed approximately 180 degrees in circadian time to depolarization. 3. Altering BRN membrane potential with ionic treatments, depolarizing with elevated K+ seawater or hyperpolarizing with lowered Na+ seawater, produces phase shifts similar to current injection. 4. The light-induced depolarization of the basal retinal neurons is necessary for phase shifts by light. Suppressing the light-induced depolarization with injected current inhibits light-induced phase shifts. 5. The ability of membrane potential changes to shift oscillator phase is dependent on extracellular calcium. Reducing extracellular free Ca++ from 10 mM to 1.3 X 10(-7) M inhibits light-induced phase shifts without blocking the photic response of the BRNs. The results indicate that changes in the membrane potential of the pacemaker neurons play a critical role in phase shifting the circadian rhythm, and imply that a voltage-dependent and calcium-dependent process, possibly Ca++ influx, shifts oscillator phase in response to light.     

Effects of ultraviolet radiation on the tension and the cyclic GMP level of bovine mesenteric arteries

Strips of bovine mesenteric arteries brought to sustained contraction by the addition of 3.0 microM phenylephrine relaxed when exposed to ultraviolet radiation (366 nm). The relaxation was reversible and associated with a rapid increase in the cGMP content. After termination of the radiation the cGMP level rapidly decreased below the basal level. The crude soluble guanylate cyclase from the artery was stimulated about 8-fold by ultraviolet radiation (366 nm). Neither the cGMP-phosphodiesterase activity nor the cAMP level were found to be changed during irradiation. The ultraviolet light-induced relaxation was not dependent on an intact intimal surface. Furthermore, the relaxing effect was found to be enhanced and accompanied by a larger increase of the cGMP level in nitroglycerin-tolerant arteries. The present results show that the ultraviolet light-induced relaxation in bovine mesenteric arteries is associated with a rapid increase in the cGMP content and that ultraviolet light and nitrocompounds may exert their relaxing actions through a common substance.     

X-ray diffraction and flash photolysis studies of M intermediate of lattice-contracted purple membrane

The effects of cross-linking and lattice contraction of purple membrane (PM) on the photodynamics of bacteriorhodopsin (bR) and on the tertiary structure were studied by flash photolysis and X-ray diffraction. To get a contracted lattice form of PM, native PM, and/or PM cross-linked by glutaraldehyde were treated with deoxycholate or Triton X-100. Part of the Triton-treated cross-linked PM was further incubated with Bio-Beads SM-2 to remove Triton X-100. In the modified PM, several long-lived components of the M intermediate appeared, the features of which were related to the environment of bR. Also, X-ray diffraction studies using synchrotron radiation were performed on the modified PM under intense light irradiation (lambda greater than 500 nm) in which 40-80% of bR was photoconverted to the M state. In the Triton-treated cross-linked PM dispersed in 0.25% Triton X-100, the unit cell of membrane crystalline lattice was enlarged from 58.8 to 59.8 A and the crystalline order decreased with irradiation. The analysis of X-ray diffraction patterns suggests that light-induced conformational changes of bR correlated with the Triton content of the environment and an increase of substitution disorder was caused by these changes, but the average location of bR was unchanged. However, the other modified PM showed no significant changes of diffraction, upon light irradiation.     

6-OHDA帕金森病大鼠快动眼睡眠状态下皮层脑电及基底节场电位的异常变化

目的：了解帕金森病（PD）模型大鼠在快动眼睡眠状态下皮层脑电和基底节场电位的异常变化。方法：用6-羟基多巴胺（6-OHDA）脑内两点注射法建立PD大鼠模型,并经阿扑吗啡注射诱发旋转对模型进行评价。通过多导宏电极在体电生理记录技术结合视频录像,对正常大鼠和6-OHDA大鼠PD模型进行苍白球场电位和皮层M1、M2区脑电的多部位24小时同时记录。功率谱分析和相干分析用于揭示快动眼睡眠状态下各记录位点信号的频率成分以及不N记录位点神经元集群之间的变化。结果：与正常大鼠相比,6-OHDA帕金森病模型大鼠在REM期间的皮层脑电在臼和y频段上都有变化：初级运动皮质M1区的θ频段成分消失,辅助运动区M2的θ频段成分略有增加,患侧苍白球的θ频段成分增大显著;M1区的γ频段成分增大,而γ频段成分在苍白球基本没有变化。结论：6-OHDA对中脑多巴胺能神经元的损害可造成大鼠双侧皮层M1区θ节律的消失和γ节律的增强,以及对侧M1-M2区之间在γ节律上的同步被显著增强,而γ节律在苍白球没有变化。这些异常电活动可能是由于VTA受损引起从而与帕金森病的快动眼睡眠行为障碍有关。