Synchronization and signal transmission in protoplasmic strands of Physarum

The importance of endoplasmic streaming in the synchronization of contraction activites in plasmodial strands of Physarum was investigated under experimental conditions allowing simultaneous observation of the endoplasmic flow in the middle part of a strand mounted as a trapeze and the measurement of isometric contraction activities of the arms of the trapeze, as well as of the activities of the strand portion connecting the arms. The correlation of longitudinal and radial contraction activities in different regions of a trapeze was examined. Whereas the arms and the middle part of a trapeze contract synchronously in a longitudinal direction (in-phase behaviour), an antiphase correlation appeared when comparing the longitudinal contraction activity of the arms and the radial activity of the middle part. This result is interpreted to mean that the middle part is able to perform isotonic contractions which induce radial dilatation of the strands. No clear-cut correlation between longitudinal and radial activities could be found when measuring simultaneously both activities in one and the same arm of a trapeze by combining tensiometry and cinematography. Protoplasmic shuttle streaming within a strand mounted as a trapeze is found to run regularly out of one arm through the middle part into the other arm, and vice versa. There is no correlation between the time points of streaming reversal and a certain stage of contraction cycles as presented by the contraction curves of the arms. However, there is a good correlation between the points of streaming reversal and the phase deviations of the longitudinal contraction activities of the arms. The importance of these phase deviations for the control of streaming reversal, i.e., for the generation of hydrostatic pressure differences in a system working with phase synchrony, is discussed. The role of endoplasmic streaming as a pacemaker for synchronization phenomena of contraction activities is stressed. The possibility is discussed that shuttle streaming of endoplasm acts as a mechanical coupling within the regulation phenomena resulting in spatial monorhythmicity.Partly presented at the Cell Motility workshop, 8^th European Muscle Conference, Heidelberg 17.–20. September 1979     

Thermotaxis and protoplasmic oscillations inPhysarum plasmodia analysed in a novel device generating stable linear temperature gradients

Summary The application of sublethal temperature gradients offers a simple, non-invasive means for in vivo studies of thermotaxis and other temperature-dependent processes in various organisms. Development, for instance, can be dramatically desynchronized, and the resulting development gradients allow to analyze physiological inter-dependencies between locally separated subsystems. For this purpose a simple device has been developed, by which a stable linear gradient of 8 °C/cm is established on an inert metal sheet with the aid of Peltier elements. The effects of linear temperature gradients on fusion, growth, and migration of plasmodia of the slime moldPhysarum polycephalum was filmed by 16 mm film time-lapse technique, and their local contraction—relaxation cycles analysed by multistrip kymography, which represents a graphic documentation of the spatio-temporal pattern of protoplasmic movements that occur along well-defined regions within the giant cell.Physarum plasmodia preferentially fuse, and grow, in the range of 24–26 °C. Different parts of a single macroplasmodium can simultaneously show positive and negative thermotaxis. The contraction—relaxation cycles generating the protoplasmic shuttle streaming within the network of veins essentially depend on local temperatures and are instantaneously desynchronized by the temperature gradient. Thus they cannot be controlled by a central pacemaker or an overall electric signal. However, there is a strong tendency to locally synchronize the various oscillation frequencies present within the giant cell if temperature differences do not exceed 2 °C.     

Propagated waves induced by gradients of physiological factors within plasmodia ofPhysarum polycephalum

Plasmodia ofPhysarum polycephalum were analyzed with the aid of cinematography and the infrared reflection technique for characterization of the phase behavior of their oscillating contraction activity, with special emphasis placed on the effects of temperature gradients. In response to temperature gradients, phase gradients were documented cinematographically as well as by infrared registration. A quantitative evaluation of the cinematographically recorded phenomena was carried out with the aid of streak photography. The phase gradient is directed across the region of the temperature gradient with a delay in phase toward the colder side. The correspondingly generated waves are as short as 1 mm and are propagated toward the colder region. A comparison of these waves with the known flickering phenomena in cinematographic films reveals a common nature of both.Supported by Deutsche Forschungsgemeinschaft.     

Repair of functional and ultrastructural alterations after thermal injury of Physarum polycephalum

Repair of thermal injury of Physarum polycephalum Schw. plasmodia has been studied by light and electron microscopy. As a result of heating the plasmodia for 10 min at 42°C both the unordered and shuttle protoplasmic streaming were arrested; the outer plasmodial membrane showed alterations at sites of contact with water; the onset of the next mitosis was considerably delayed. The plasmodial ultrastructure was markedly disturbed, including disappearance of the granular component of the nucleoili and a compact, almost fibrillar structure of the latter. The mitochondria became distorted and their intracristal spaces enlarged while the outer and inner membranes appeared in some places to be separated. Glycogen particles disappeared from the cytoplasm. Recovery of both types of protoplasmic streaming of the motility of the plasmodium, of the resistance of its membrane to contact with water, and of the ability of the organism to go through the cell cycle went all hand in hand with the normalization of the structure of nucleoli, mitochondria and cytoplasm. All of the functional and structural characteristics are normalized within ca. 9 h following heating.     

Body temperature and behaviour of golden hamsters (Mesocricetus auratus) and ground squirrels (Spermophilus tridecemlineatus) in a thermal gradient

Golden hamsters and thirteen-lined ground squirrels were maintained individually in a thermal gradient (14°C to 33°C) for several weeks under a 14L: 10D light-dark cycle. Animals of both species showed robust daily rhythms of body temperature and locomotor activity with acrophases consistent with the habits of the species (diurnal acrophases in the diurnal squirrels and nocturnal acrophases in the nocturnal hamsters). Hamsters showed a robust daily rhythm of temperature selection 180° out of phase with the rhythms of body temperature and locomotor activity. Squirrels did not show a daily rhythm of temperature selection. These results raise the hypothesis that a daily rhythm of temperature selection is exhibited by nocturnal but not by diurnal endotherms.     

Behavioral response of the New England dog whelk,Nassarius trivittatus,to a temperature gradient

Thirty New England dog whelks (Mollusca: Gastropoda: Nassariidae), Nassarius trivittatus, were tested as a group for 3 days in a long linear gradient trough encompassing a thermal gradient ranging from 5°C to 45°C. The modal thermal preferendum of the snails lay between 30°C and 35°C. The lowest temperature interval voluntarily occupied by the snails was 15–20°C, the highest 35–40°C. W e believe this to be the first published study of temperature preference and avoidance behavior of gastropod molluscs.     

Centriole maturation in the amoebae ofPhysarum polycephalum

Summary The precise geometry of pro-centriole formation has been studied inPhysarum polycephalum amoebae. The spatial references used were the posterior and the anterior kinetosomes which are unequivocally defined by the presence of the posterior para-kinetosomal structure, the microtubular array 4 and the microtubular arrays 1, 2, and 3. The observations made suggest that pro-centrioles follow a maturation process. A pro-centriole formed during the n^th cell cycle becomes the posterior kinetosome during the (n + 1)^th cell cycle and the anterior one during all the following cell cycles. Pro-centriole formation occurs late in the cell cycle. This observation disagrees with a role of pro-centriole formation in the regulation of S phase in contrast to what has been suggested in other eucaryotic cells.     

The physiological response of Northern krill (Meganyctiphanes norvegica) to temperature gradients in the Kattegat

The Alkor-Deep (140 m), which forms part of a depression system in the northern Kattegat channel east of the island of Læsø (Denmark), is the location of a self sustaining population of Northern krill, Meganyctiphanes norvegica (Euphausiacea). This population is exposed to one of the most pronounced thermal gradients within the distributional range of this pelagic crustacean. During summer, the temperature of the water column ranges between 4 and 6 in the deep to 16 °C near the surface which results in the krill being exposed to temperature differences of 8–10 °C during diel vertical migration. Oxygen consumption rates were used to investigate the physiological adaptation of the animal to such gradients in temperature. The rates were found to increase exponentially from 31 mol O₂ h^-1 g_dw ^-1 at 4 °C to 72 mol O₂ h^-1 g_dw ^-1 at 16 °C, giving a Q ₁₀-value of 2.0, and indicating that physiological adaptation to varying thermal conditions does not take place. Behavioural adaptations are discussed which may help the krill to cope with large temperature gradients in their environment.     

Evaporative cooling and endothermy in the 13-year periodical cicada,Magicicada tredecem (Homoptera: Cicadidae)

Summary The thermobiology of a cicada, Magicicada tredecem, from a warm, high humidity environment was investigated. Thoracic temperature (T_th) of M. tredecem in the field was strongly dependent on, and consistently higher than, ambient temperature (T_am), averaging 33.0±0.19°C on warm sunny days (T_am=28–29°C, rh=60–75%). Laboratory studies documented cuticle water fluxes high enough ( 5mg · cm^–2 · h^–1 in dry air at 40°C) to result in a significant degree of passive evaporative cooling, but the ability of M. tredecem to actively facilitate evaporative water loss during thermal stress is comparatively limited: water loss rates (WLR) of live M. tredecem at 40°C (dry air) were only 35–45% greater than those of dead cicadas. The limited ability of M. tredecem to facilitate transcuticular WLR is associated with limited surface distribution of the cuticular ducts through which water is actively extruded during evaporative cooling. In the laboratory, active extrusion of water had no appreciable effect on T_th, demonstrating that evaporative cooling was due largely to passive water flux through the highly permeable cuticle. The location of the abdominal pore tracts is such that extrusion of water through the ducts may preferentially cool the heart and perhaps other abdominal tissues. Long-term climatological data indicate that M. tredecem rarely encounters T_am levels high enough (i.e., above its apparent T_th setpoint of 34–35°C) to require evaporative cooling. Inactive M. tredecem can endothermically increase T_th. An hypothesis is proposed to account for the diversity of body temperature setpoints in cicadas.Abbreviations rh relative humidity - SOT standard operating temperature - T _am ambient temperature - T _b body temperature - T _sp body temperature setpoint - T _th thoracic temperature - TWF transcuticular water flux - WLR water loss rate     

Enrichment of fibrillar cytoplasmic actomyosin in protoplasmic strands of Physarum polycephalum for the production of cell-free models

Summary The treatment of isolated protoplasmic strands of Physarum polycephalum with 2.5% ethanol in a physioogical salt solution under isometric conditions induces the formation of a large amount of mostly longitudinally organized actomyosin fibrils in the endoplasmic channel, a region normally free of actomyosin fibrils. The quantity of fibrillogenesis as well as the concomitant force output during the induced contractures are dependent on the Ca^{+ +}- content and the temperature of the test solution. The method was developed to optimize the structure of the plasmodial strands before their subsequent transformation into cell-free models by permeabilization and extraction of the strands.Cryosections of plasmodial strands containing cytoplasmic actomyosin fibrils stained with fluorescently labeled phallotoxins offer a further assay for the study of their contraction physiology under cell-free conditions.     

Influence of Thermoperiod on Growth and Development in Cucumber

We studied the influence of gradient temperature regimes on various parameters of the formation of shoots and roots of cucumber plants, such as rate of leaf appearance, rate of growth, duration of growth and length of leaves, and the rate of growth shoots organs and roots. The plants were grown under the controlled conditions: at different combinations of day and night temperature, illumination 100 W/m², and 12 h photoperiod. The comparison of constant and fluctuating diurnal temperature regimes has shown that in the optimal area for all studied indices, the highest values were recorded at the constant daily temperature (25°C for all growth indices of shoots and 20°C for growth of roots), while all gradient regimes either did not affect, or exerted inhibitory effects on the plant. Outside the optimum area, the effects of gradient temperatures differed. The main acting fluctuating temperatures, that exerted stimulating effects, combined low hardening (15°C) and optimal temperatures (25°C), which was earlier described for animals. The 15/35 and 35/15°C combinations were unambiguously inhibitory, since both temperatures are hardening for the cucumber. A lesser stimulating effect of gradient temperatures on the developmental rate in a plant, as compared to poikilothermic animals, could be due to a greater autonomy of plant ontogenesis because of autotrophy and, correspondingly, a greater degree of homeostasis. The mechanisms accounting for the responses to temperature gradients are similar in different groups of ectotherms.     

Thermal preference and activity thresholds in populations of Argia vivida (Odonata: Coenagrionidae) from habitats with different thermal regimes

The hypothesis was tested that isolated populations of Argia vivida Hagen living in habitats with different thermal regimes would show similar larval temperature preferenda, similar distributions in a temperature gradient, similar larval upper temperature acitivity thresholds, and similar adult minimum temperature flight thresholds. The hypothesis was supported in all cases, except for distribution within the gradient, where there were significantly fewer observations below 22°C in a population from a habitat with a fluctuating diel and annual temperature regime than in a population from a more thermally stable habitat. Larval modal temperature preferendum was 28°C; escape temperature (EST) was 35.4–36.4°C, critical thermal maximum (CTM) was 39.1–41.0°C, and upper lethal temperature (ULT) was 44.4–46.0°C. While technical difficulties affected the estimates of flight thresholds, there was no difference between the field estimates from different sites. Minimum body temperature for flight appears to be about 25°C, apparently higher than for several other zygopterans, while larval activity thresholds and 96 hr LD50 of 36.8°C are similar to those recorded for other odonates.     

Temperature preferences and tolerances of three fish species inhabiting hyperthermal ponds on mangrove islands

The fish species Cyprinidon artifrons, Floridichthys carpio, and Gambusia yucatana inhabit shallow mangrove ponds off the coast of Belize. Portions of these ponds experience a diurnal temperature change from 26 °C at night to 40 °C and above during midday. Repeated field observations indicate Cyprinidon prefer the warmer (and much larger) portions of the ponds whereas the other two species stay in the cooler areas.The hypothesis that temperature is serving as a cue for partitioning within the ponds was supported by laboratory thermal gradient tests in which Cyprinidon preferred temperatures clearly higher than the other two species.The critical thermal maximum (CTM) was determined for the three species using members that had been acclimated to either a daily cycling temperature similar to that for the ponds, or to the mean of the 24-hour cycle (30 °C). Cyprinidon acclimated to the cycling temperature had a CTM of 45.5 °C, which apparently sets a new record for fish CTM. Acclimation to a constant 30 °C lowered the CTM to 43.7 °C. Floridichthys and Gambusia acclimated to the cycled temperature had CTMs of 43.9 and 43.3 °C respectively, and 42.5 and 42.6 °C for those acclimated to 30 °C.All three species appear to have the ability to tolerate the high temperatures throughout the ponds but only Cyprinidon utilize the whole pond during the day. This may help to explain the large populations of Cyprinodon found in these mangrove ponds compared to the other species.     

Cylindrospermopsin production and release by the potentially invasive cyanobacterium Aphanizomenon ovalisporum under temperature and light gradients

The growth rates, production and release of the potent cytotoxin cylindrospermopsin (CYN) were studied in batch and semi-continuous cultures of Aphanizomenon ovalisporum (Cyanobacteria; Nostocaceae) strains UAM 289 and UAM 290 from Spain, over a gradient of temperatures (10–40 °C) and irradiances (15–340 μE m⁻² s⁻¹). This species grew in temperatures ranging from 15 °C to 35 °C as well as under all irradiances assayed. The growth rates ranged from 0.08 d⁻¹ to 0.35 d⁻¹, and the maximum growth was recorded above 30 °C and at 60 μE m⁻² s⁻¹. CYN was produced under all conditions where net growth occurred. Total CYN reached up to 6.4 μg mg⁻¹ dry weight, 2.4 μg mm⁻³ biovolume, 190.6 fg cell⁻¹ and 0.5 μg μg⁻¹ chlorophyll a. Although CYN concentrations varied only 1.9-fold within the 15–30 °C range, a drastic 25-fold decrease was observed at 35 °C. The irradiance induced up to 4-fold variations, with maximum total CYN measured at 60 μE m⁻² s⁻¹. An elevated extracellular CYN share ranging from 20% to 35% was observed during the exponential growth phase in most experiments, with extreme temperatures (15 and 35 °C) being related to the highest release (63% and 58%, respectively) and without remarkable influence of irradiance. Growth did not have a direct influence on either CYN production or release throughout the entire range of experimental conditions. Our study demonstrates a strong and stable production and release of CYN by A. ovalisporum along field-realistic gradients of temperature and light, thus becoming a predictive tool useful for the management of water bodies potentially affected by this ecologically plastic cyanobacterium.     

Behavioral response of the starfish asterias forbesh to a thermal gradient

ThirtyAsterias fbrbesii (Echinodermata: Asteroidea: Asteriidae) were tested as a group for 3 days in a long linear trough with a thermal gradient encompassing 5–45°C. The modal thermal preferendum of the starfish was between 20 and 25°C. The starfish avoided temperatures below 15°C or above 35°C.     

Respiration and heat production by the inflorescence of Philodendron selloum Koch

During a 2-d sequence of anthesis, the spadices of the thermogenic arum lily, Philodendron selloum, regulated maximum temperature within a small range (37–44°C) by reversible thermal inhibition of respiratory heat production. This response protects the inflorescence and the attracted insects from thermal damage. Heat production by whole spadices, measured by O₂ respirometry, equalled heat loss, measured by gradient layer calorimetry, which confirmed the heat equivalence of O₂ consumption (20.4 J ml^-1). This also indicated that there was no net phosphorylation during thermogenesis, heat production being the primary function of high rates of respiration. The sterile male florets consumed about 30 ml g^-1 h^-1 and the average 124-g spadix produced about 7 W to maintain a 30°C difference between spadix and ambient temperature. Most of the energy for thermogenesis is present in the florets before anthesis. Despite high respiratory rates, thermogenesis is an energetically inexpensive component of the reproductive process.     

An improved breeding method for solitarious locusts

In order to unravel the physiological, endocrine, and behavioral differences between gregarious and solitarious forms of the desert locust, Schistocerca gregaria (Forsk.) (Orthoptera, Acrididae), a constant supply of rather large numbers of solitary individuals has to be guaranteed. This represents a bottleneck, mainly because of the intensity of the labor involved and limited laboratory accommodation. The method we describe here substantially reduces the space and manpower needed. The survival rate we obtained in the solitarised population was relatively high, reaching about 55%. The optimal rearing temperature proved to be 32–36 °C. Cabbage leaves and oat flakes sufficed for feeding all year round. Special racks have been designed that enable high density stacking and easy handling. The solitarisation process was monitored over ten consecutive generations. Changes in morphometrics, eye stripes, color, and behavior were recorded, of which only morphometrics, temperature related development, and mortality are discussed. A shift towards the solitarious phase was recorded, with clear differences between gregarious, 1^st generation and 7^th to 10^th generation solitarious locusts.     

Variable thermal sensitivity as output of a circadian clock controlling the bimodal rhythm of temperature choice in the ant Camponotus mus

Along a thermal gradient and under a LD 1212 h cycle, nurse workers of the ant Camponotus mus select for the brood two different temperatures daily: 30.8°C at the middle of the light period (circadian phase = 90°), and 27.5°C 8 h later, during the dark period (CP = 210°). Brood-carrying activity proved to be self-sustained, running its two daily bursts free with a similar period of 23.5 h, under both LL and DD. The LD alternation acted as a strong Zeitgeber. A phase-delay of the LD 1212 h cycle reset the overt rhythm at once, being both daily events locked-on to the delayed light: dark transition. However, changes in expression, non-occurrence, or even splitting of the two daily brood-carrying events during resetting depended on the phase of the delayed DL transition. By comparing the occurrence of activity with predictions based on a threshold curve of thermal sensitivity, results indicated that an immediate resetting of the involved pacemaker actually takes place. Nurse workers do not directly control the total time spent by the brood at the selected temperature. Instead, the endogenously-driven thermal sensitivity triggers their thermal-searching behavior at two critical times of the day, when environmental temperature is expected to reach its maximum and minimum.     

Isolation and characterization of an obligately anaerobic,polysaccharolytic, extremely thermophilic member of the genus Spirochaeta

An obligately anaerobic, extremely thermophilic Spirochaeta species was isolated from a thermal spring on the edge of Green Lake on Raoul Island of the Kermadec archipelago. The strain, designated RI 19.B1 (=DSM 6192) displayed the morphological characteristics typical of Spirochaeta species: regularly coiled long thin cells consisting of a crenulated outer sheath surrounding a central coiled protoplasmic cylinder. Between the outer sheath and the protoplasmic cylinder were two longitudinal periplasmic fibrils in a 1-2-1 arrangement, each anchored by an insertion disc near one pole, whereas the other end was not anchored. The strain displayed a strictly anaerobic, chemoorganotrophic, fermentative metabolism and was able to grow on a variety of mono-, di- and polysaccharides, including cellulose. Sugar alcohols, organic and amino acids were not utilized. Growth supplements were not required, but CO₂ was required to produce consistent growth. Strain RI 19.B1 had temperature, pH and salinity optima of 64–66°C, pH 6.95 and 0.4% NaCl respectively. The maximum growth temperature and salinity were 73°C and 2.5% respectively. Glucose was fermented to lactate, acetate, carbon dioxide, and hydrogen. Succinate, ethanol and formate were not detected. The strain displayed the resistance to rifampicin typical of Spirochaeta species. The mol % G+C of DNA from strain RI 19.B1 was 52%.