Mutants of thermotaxis in Dictyostelium discoideum

Amoebae of Dictyostelium discoideum, strain HL50 were mutagenized with N-methyl-N′-nitro-N-nitrosoguanidine, cloned, allowed to form pseudoplasmodia and screened for aberrant positive and negative thermotaxis. Three types of mutants were found. Mutant HO428 exhibits only positive thermotaxis over the entire temperature range (no negative thermotaxis). HO596 and HO813 exhibit weakened positive thermotaxis and normal negative thermotaxis. The weakened positive thermotactic response results in a shift toward warmer temperatures in the transition temperature from negative to positive thermotaxis. Mutant HO209 exhibits weakened positive and negative thermotactic responses and has a transition temperature similar to the ‘wild type’ (HL50). The two types of mutants represented by HO428, HO596 and HO813 support the model that positive and negative thermotaxis have separate pathways for temperature sensing. The type of mutants which contains HO209 suggests that those two pathways converge at some point before the response.     

Serotonin Control of Thermotaxis Memory Behavior in Nematode Caenorhabditis elegans

Caenorhabditis elegans is as an ideal model system for the study of mechanisms underlying learning and memory. In the present study, we employed C. elegans assay system of thermotaxis memory to investigate the possible role of serotonin neurotransmitter in memory control. Our data showed that both mutations of tph-1, bas-1, and cat-4 genes, required for serotonin synthesis, and mutations of mod-5 gene, encoding a serotonin reuptake transporter, resulted in deficits in thermotaxis memory behavior. Exogenous treatment with serotonin effectively recovered the deficits in thermotaxis memory of tph-1 and bas-1 mutants to the level of wild-type N2. Neuron-specific activity assay of TPH-1 suggests that serotonin might regulate the thermotaxis memory behavior by release from the ADF sensory neurons. Ablation of ADF sensory neurons by expressing a cell-death activator gene egl-1 decreased the thermotaxis memory, whereas activation of ADF neurons by expression of a constitutively active protein kinase C homologue (pkc-1(gf)) increased the thermotaxis memory and rescued the deficits in thermotaxis memory in tph-1 mutants. Moreover, serotonin released from the ADF sensory neurons might act through the G-protein-coupled serotonin receptors of SER-4 and SER-7 to regulate the thermotaxis memory behavior. Genetic analysis implies that serotonin might further target the insulin signaling pathway to regulate the thermotaxis memory behavior. Thus, our results suggest the possible crucial role of serotonin and ADF sensory neurons in thermotaxis memory control in C. elegans.     

Distinct thermal migration behaviors in response to different thermal gradients in Caenorhabditis elegans

The nematode Caenorhabditis elegans exhibits a complex behavior called thermotaxis in response to temperature. This behavior is defined as a form of associative learning, in which temperature pairs with the presence or absence of food. Different interpretations have been drawn from the diverse results obtained by several groups, mainly because of the application of different methodologies for the analysis of thermotaxis. To clarify the discrepancies in behavioral observations and subsequent interpretations by different laboratories, we attempted to systematize several parameters to observe thermotaxis behavior as originally defined by Hedgecock and Russell in 1975. In this study, we show clearly how C. elegans can show a conditioned migration toward colder or warmer areas on a thermal gradient, given certain criteria necessary for the observation of thermotaxis. We thus propose to distinguish thermotaxis from other temperature-related behaviors, such as the warm avoidance response displayed at temperature gradients of 1°C/cm and steeper.     

High Concentration of Vitamin E Decreases Thermosensation and Thermotaxis Learning and the Underlying Mechanisms in the Nematode Caenorhabditis elegans

α-tocopherol is a powerful liposoluble antioxidant and the most abundant isoform of vitamin E in the body. Under normal physiological conditions, adverse effects of relatively high concentration of vitamin E on organisms and the underlying mechanisms are still largely unclear. In the present study, we used the nematode Caenorhabditis elegans as an in vivo assay system to investigate the possible adverse effects of high concentration of vitamin E on thermosensation and thermotaxis learning and the underlying mechanisms. Our data show that treatment with 100–200 µg/mL of vitamin E did not noticeably influence both thermosensation and thermotaxis learning; however, treatment with 400 µg/mL of vitamin E altered both thermosensation and thermotaxis learning. The observed decrease in thermotaxis learning in 400 µg/mL of vitamin E treated nematodes might be partially due to the moderate but significant deficits in thermosensation, but not due to deficits in locomotion behavior or perception to food and starvation. Treatment with 400 µg/mL of vitamin E did not noticeably influence the morphology of GABAergic neurons, but significantly decreased fluorescent intensities of the cell bodies in AFD sensory neurons and AIY interneurons, required for thermosensation and thermotaxis learning control. Treatment with 400 µg/mL of vitamin E affected presynaptic function of neurons, but had no remarkable effects on postsynaptic function. Moreover, promotion of synaptic transmission by activating PKC-1 effectively retrieved deficits in both thermosensation and thermotaxis learning induced by 400 µg/mL of vitamin E. Therefore, relatively high concentrations of vitamin E administration may cause adverse effects on thermosensation and thermotaxis learning by inducing damage on the development of specific neurons and presynaptic function under normal physiological conditions in C. elegans.     

Onchocerca volvulus: biochemical and morphological characteristics of the surface of third- and fourth-stage larvae

The annulated cuticles of third- and fourth-stage larvae of Onchocerca volvulus have the typical structure of other nematodes but the cuticle of fourth-stage larvae was thinner. The surface of the third-stage larva was wrinkled and fuzzy, while that of the fourth-stage was smooth. Intermediate stages in the formation of the new cuticle and epicuticle beneath the old basal layer and of the separation of the cuticles are shown. Monoclonal antibodies specific to the surface of third-stage larvae did not react with the surface of the fourth-stage larvae. Binding of the monoclonal antibodies to the third-stage larvae was abrogated by treatment of the worms with trypsin and proteinase K, but was unaffected by treatment with periodate or the detergents sodium deoxycholate and SDS. The lectins RCA120 and WGA, but not any of the other lectins tested, bound only to the surface of fourth-stage larvae, and not to that of third-stage larvae. The surfaces of third- and fourth-stage larvae were shown to be different and contained stage-specific surface epitopes.     

Sensitivity of Paramecium Thermotaxis to Temperature Change

SYNOPSIS. The relationship to swimming velocity of the critical temperature gradient necessary for inducing thermotaxis in Paramecium caudatum was analyzed at various temperatures and viscosities. Since the critical temperature gradient was linearly proportional to the inverse of the swimming velocity, it is concluded that P. caudatum detects temperature changes by locomotion through space and thus exhibits thermotaxis, provided the rate of change is > 0.055 C/sec. The swimming velocity jump was observed when the ciliates were subjected to a stepwise temperature change toward an optimum with a rate > 0.05 C/sec; the jump was not observed, however, when they were subjected to a change toward an unpreferred temperature with the same rate. Hence, thermotaxis can be explained partly by the swimming velocity jump brought about when the cells are swimming toward an optimum temperature in a spatial gradient. It is suggested that thermotaxis might be a direct manifestation of the dynamic properties of membrane as a receptor.     

Identification of guanylyl cyclases that function in thermosensory neurons of Caenorhabditis elegans

The nematode Caenorhabditis elegans senses temperature primarily via the AFD thermosensory neurons in the head. The response to temperature can be observed as a behavior called thermotaxis on thermal gradients. It has been shown that a cyclic nucleotide-gated ion channel (CNG channel) plays a critical role in thermosensation in AFD. To further identify the thermosensory mechanisms in AFD, we attempted to identify components that function upstream of the CNG channel by a reverse genetic approach. Genetic and behavioral analyses showed that three members of a subfamily of gcy genes (gcy-8, gcy-18, and gcy-23) encoding guanylyl cyclases were essential for thermotaxis in C. elegans. Promoters of each gene drove reporter gene expression exclusively in the AFD neurons and, moreover, tagged proteins were localized to the sensory endings of AFD. Single mutants of each gcy gene showed almost normal thermotaxis. However, animals carrying double and triple mutations in these genes showed defective thermotaxis behavior. The abnormal phenotype of the gcy triple mutants was rescued by expression of any one of the three GCY proteins in the AFD neurons. These results suggest that three guanylyl cyclases function redundantly in the AFD neurons to mediate thermosensation by C. elegans.     

Genetic control of temperature preference in the nematode Caenorhabditis elegans

Animals modify behavioral outputs in response to environmental changes. C. elegans exhibits thermotaxis, where well-fed animals show attraction to their cultivation temperature on a thermal gradient without food. We show here that feeding-state-dependent modulation of thermotaxis is a powerful behavioral paradigm for elucidating the mechanism underlying neural plasticity, learning, and memory in higher animals. Starved experience alone could induce aversive response to cultivation temperature. Changing both cultivation temperature and feeding state simultaneously evoked transient attraction to or aversion to the previous cultivation temperature: recultivation of starved animals with food immediately induced attraction to the temperature associated with starvation, although the animals eventually exhibited thermotaxis to the new temperature associated with food. These results suggest that the change in feeding state quickly stimulates the switch between attraction and aversion for the temperature in memory and that the acquisition of new temperature memory establishes more slowly. We isolated aho (abnormal hunger orientation) mutants that are defective in starvation-induced cultivation-temperature avoidance. Some aho mutants responded normally to changes in feeding state with respect to locomotory activity, implying that the primary thermosensation followed by temperature memory formation remains normal and the modulatory aspect of thermotaxis is specifically impaired in these mutants.     

Construction of a thermotaxis chamber providing spatial or temporal thermal gradients monitored by an infrared video camera system.

A thermotaxis chamber was constructed to quantitatively study thermotaxis in eukaryotic amoeboid cells. The apparatus provided either spatial or temporal temperature gradients in an observation chamber set in an inverted microscope. With an infrared video camera system, spatial thermal gradients were monitored directly and the temperature at the actual location of the cells could be estimated accurately. This enabled a precise determination of the strength of thermal stimuli. With this apparatus, we were able to simultaneously measure temperature and observe cellular behavior directly. This feature permits quantitative studies on stimulus-response relationships. The utility of the apparatus was demonstrated by thermotaxis assay under a spatial thermal gradient in polymorphonuclear leukocytes. Since this apparatus can also provide temporal thermal gradients, it may have several applications in studies of temperature-dependent phenomena in cell biology.     

Copine A is expressed in prestalk cells and regulates slug phototaxis and thermotaxis in developing Dictyostelium

Copines are calcium-dependent membrane-binding proteins found in many eukaryotic organisms. We are studying the function of copines using the model organism, Dictyostelium discoideum. When under starvation conditions, Dictyostelium cells aggregate into mounds that become migrating slugs, which can move toward light and heat before culminating into a fruiting body. Previously, we showed that Dictyostelium cells lacking the copine A (cpnA) gene are not able to form fruiting bodies and instead arrest at the slug stage. In this study, we compared the slug behavior of cells lacking the cpnA gene to the slug behavior of wild-type cells. The slugs formed by cpnA- cells were much larger than wild-type slugs and exhibited no phototaxis and negative thermotaxis in the same conditions that wild-type slugs exhibited positive phototaxis and thermotaxis. Mixing as little as 5% wild-type cells with cpnA- cells rescued the phototaxis and thermotaxis defects, suggesting that CpnA plays a specific role in the regulation of the production and/or release of a signaling molecule. Reducing extracellular levels of ammonia also partially rescued the phototaxis and thermotaxis defects of cpnA- slugs, suggesting that CpnA may have a specific role in regulating ammonia signaling. Expressing the lacZ gene under the cpnA promoter in wild-type cells indicated cpnA is preferentially expressed in the prestalk cells found in the anterior part of the slug, which include the cells at the tip of the slug that regulate phototaxis, thermotaxis, and the initiation of culmination into fruiting bodies. Our results suggest that CpnA plays a role in the regulation of the signaling pathways, including ammonia signaling, necessary for sensing and/or orienting toward light and heat in the prestalk cells of the Dictyostelium slug.     

Simulation of C. elegans thermotactic behavior in a linear thermal gradient using a simple phenomenological motility model

The nematode Caenorhabditis elegans has been reported to exhibit thermotaxis, a sophisticated behavioral response to temperature. However, there appears to be some inconsistency among previous reports. The results of population-level thermotaxis investigations suggest that C. elegans can navigate to the region of its cultivation temperature from nearby regions of higher or lower temperature. However, individual C. elegans nematodes appear to show only cryophilic tendencies above their cultivation temperature. A Monte-Carlo style simulation using a simple individual model of C. elegans provides insight into clarifying apparent inconsistencies among previous findings. The simulation using the thermotaxis model that includes the cryophilic tendencies, isothermal tracking and thermal adaptation was conducted. As a result of the random walk property of locomotion of C. elegans, only cryophilic tendencies above the cultivation temperature result in population-level thermophilic tendencies. Isothermal tracking, a period of active pursuit of an isotherm around regions of temperature near prior cultivation temperature, can strengthen the tendencies of these worms to gather around near-cultivation-temperature regions. A statistical index, the thermotaxis (TTX) L-skewness, was introduced and was useful in analyzing the population-level thermotaxis of model worms.     

Effect of stimulus strength and adaptation on the thermotactic response of Dictyostelium discoideum pseudoplasmodia

The thermotactic responses of Dictyostelium discoideum strain HL50 and mutants derived from this strain have been characterized by curves of stimulus-strength vs response. With gradient midpoint temperatures of 16 and 24 °C, these curves are typical of those of a single response, i.e., the strength of the response increases with increasing stimulus strength until at some strength the response saturates. However, with a gradient midpoint temperature close to the transition from negative to positive thermotaxis, the sign of the thermotactic response depends on gradient strength. These observations support the hypothesis that the transduction pathways for positive and negative thermotaxis act concurrently and contain separable elements. An investigation of the adaptation of thermotaxis indicated that the stimulus-strength-dependence and midpoint-temperature-dependence of both thermosensory responses was altered by shifting the growth and development temperature.     

Steepness of thermal gradient is essential to obtain a unified view of thermotaxis in C. elegans

One of the adaptive behaviors of animals in their environment is thermotaxis, by which they migrate toward a preferred temperature. This sensorimotor integration is accomplished by choosing one of two behaviors depending on the surrounding temperature, namely thermophilic or cryophilic movement. Caenorhabditis elegans exhibits thermotaxis and its migration behavior has been analyzed experimentally at both the population and individual levels. However, some experimental data are inconsistent especially for thermophilic movement, which is expected to be observed in lower than favorable temperatures. There are no experimental analyzes that find thermophilic tendencies in the individual behavior of worms, despite multiple reports supporting thermophilic movement of the population. Although theoretical methods have been used to study thermotaxis of C. elegans, no mathematical model provides a consistent explanation for this discrepancy. Here we develop a simple biased random walk model, which describes population behavior, but which is based on the results of individual assays. Our model can integrate all previous experiments without any contradiction. We regenerate all the population patterns reported in past studies and give a consistent explanation for the conflicting results. Our results suggest that thermophilic movement is observed, even in individual movements, when the thermal gradient is sufficiently slight. On the contrary, thermophilic movement disappears when the thermal gradient is too steep. The thermal gradient is thus essential for a comprehensive understanding of the experimental studies of thermotaxis in C. elegans. Our model provides insight into an integrative understanding of the neural activity and thermotactic behavior in C. elegans.     

Thermal selection behaviour in the estuarine goby Gillichthys mirabilis Cooper

The estuarine goby Gillichthys mirabilis behaviorally thermoregulates when placed in a laboratory temperature gradient. Avoidance of temperatures above 23°C is the most evident component of this thermotaxis. Temperature preferences of non-acclimated fish do not vary significantly as a function of season; nor does temperature acclimation alter the preferendum. Varying the temperature range of the gradient apparently modifies thermotaxis. Negative phototaxis evidenced by this species is subjugated by thermal preferences. Temperature preference does not vary diurnally. The heat resistance of this species was determined; there is no apparent relationship between heat resistance and temperature preference.     

Working with what you’ve got: Changes in thermal preference and behavior in mice with or without nesting material

In laboratories mice are typically housed at ambient temperatures (T_a) of 20-24 °C, which are below their average preferred T_a of ≈30 °C. Adjusting laboratory T_a is not a solution because preferences differ depending on activity, time of the day, and gender. We tested the hypothesis that providing mice with nesting material will allow behavioral thermoregulation and reduce aversion to colder T_a. We housed C57BL/6J mice with and without nesting material in a set of 3 connected cages, each maintained at a different T_a (20, 25, or 30 °C). Mice were confined in and given free access to the T_a options to determine if thermotaxis or nest building was the primary mode of behavioral thermoregulation. As predicted, nesting material reduced aversion to 20 °C but the overall preference, in both treatments, was still 30 °C. Inactive and nesting behaviors were more likely to be seen in contact with nesting material while active behaviors were more likely to be observed when not in contact. Nest quality increased with decreasing T_a when mice could not use thermotaxis but nest quality was uncorrelated with T_a when thermotaxis was possible. Males decreased nest quality with increasing temperatures but females showed no correlation. We conclude that nesting material does not alter thermal preferences for 30 °C when thermotaxis is possible, indicating thermotaxis as the primary mode of behavioral thermoregulation. However, when thermotaxis is not possible, mice adjust nest shape depending on the T_a. Nesting material appears to partially compensate for cooler T_a and is especially important when mice are inactive. Therefore, nesting material may be a solution to the mismatch between laboratory T_a and mouse thermal preferences.     

Changes in the ultrastructure of Nematospiroides dubius (Nematoda) intestinal cells during development from fourth stage to adult

Summary The general fine structure of intestinal cells and changes which occur in ultrastructure during development from fourth-stage to adult N. dubius are reported. In fourth-stage worms pigment granules are prominent in intestinal cells. In adults the number of pigment granules appears to be reduced and phagolysosomes containing membranous profiles and pigment material increase in number. Another reorganization of cell structure involves mitochondria which are randomly distributed in the cytoplasm of cells in fourth-stage worms, concentrated in the apical cytoplasm in worms in the molting process, and confined to the base of cells in adult worms. Other changes involved structure of the nucleus, rough endoplasmic reticulum and glycogen content of cells.This investigation was supported, in part, by NIH Fellowships I-FI-GM-32750 and 5-F02-AI-32750.     

A steep thermal gradient thermotaxis assay for the nematode Caenorhabditis elegans

The nematode Caenorhabditis elegans with its well-described nervous system is one of the multicellular organisms of choice to study thermotaxis. The neuronal circuitry for thermosensation has been analyzed at the level of individual cells. Two methods have previously been described to study the behavior of C. elegans with respect to temperature: 1) isothermal tracking assays and 2) linear thermal gradients (Hedgecock and Russell, 1975). Here we present a short linear thermal gradient assay which is faster and which allows statistical evaluation of different populations using a thermotaxis index. Thin agar plates are used on which a temperature gradient from about 10 degrees to 30 degrees is induced over the distance of about 5 cm. The short linear thermal gradient uses inexpensive materials so that multiple tests can be performed in parallel in a short period of time.     

Genetic Analysis of Dictyostelium Slug Phototaxis Mutants

Mapping and complementation analysis with 17 phototaxis mutations has established 11 complementation groups phoA-phoK distributed over six linkage groups. Statistical calculations from the complementation data yielded 17 as the maximum likelihood estimate of the number of pho genes assuming all loci are equally mutable. Most of the phototaxis mutants were found to exhibit bimodal phototaxis and all were found to be impaired in positive thermotaxis supporting convergence of the photosensory and thermosensory pathways. The thermotaxis mutant HPF228 was unaltered in phototaxis suggesting that the mutation in this strain affects a gene product whose site of action is before the convergence of the two pathways. Other phenotypes such as multiple tip formation by aggregates, stumpy fruiting bodies with short or absent stalks and short migration were associated with some pho alleles suggesting multiple biological roles for some gene products important in phototransduction.     

A Mechanism for Precision-Sensing via a Gradient-Sensing Pathway: A Model of Escherichia coli Thermotaxis

Thermotaxis is the phenomenon where an organism directs its movement toward its preferred temperature. So far, the molecular origin for this precision-sensing behavior remains a puzzle. We propose a model of Escherichia coli thermotaxis and show that the precision-sensing behavior in E. coli thermotaxis can be carried out by the gradient-sensing chemotaxis pathway under two general conditions. First, the thermosensor response to temperature is inverted by its internal adaptation state. For E. coli, chemoreceptor Tar changes from a warm sensor to a cold sensor on increase of its methylation level. Second, temperature directly affects the adaptation kinetics. The adapted activity in E. coli increases with temperature in contrast to the perfect adaptation to chemical stimuli. Given these two conditions, E. coli thermotaxis is achieved by the cryophilic and thermophilic responses for temperature above and below a critical temperature T_c, which is encoded by internal pathway parameters. Our model results are supported by both experiments with adaptation-disabled mutants and the recent temperature impulse response measurements for wild-type cells. T_c is predicted to decrease with the background attractant concentration. This mechanism for precision sensing in an adaptive gradient-sensing system may apply to other organisms, such as Dictyostelium discoideum and Caenorhabditis elegans.     

The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): The parasitic stages

Smales L. R. 1977. The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): the parasitic stages. International Journal for Parasitology7: 457–461. Labiostrongylus eugenii infective larvae ingested by the Kangaroo Island Wallaby exsheathe on the saccular stomach wall, then invade the mucosa causing one or two chronic irritative hyperpastic nodules, each containing a number of larvae. Six to eight weeks later the larvae leave the stomach wall and moult to fourth-stage. The fourth-stage larva is characterized by eight lips, while the oesophagus and anterior intestine assume the adult form. The posterior ends of late fourth-stage larvae become sexually differentiated. Moulting to adult occurs after 6–8 weeks with the first gravid females being observed 3 months later. The prepatent period appears to last about 6 months. Reasons for failure of experimental infection trials and the epidemiological significance of the life cycle were discussed.