Inhibitory effects of caffeine on gustatory plasticity in the nematode Caenorhabditis elegans.

The effects of caffeine on salt chemotaxis learning were investigated using the nematode Caenorhabditis elegans. To estimate the degree of salt chemotaxis learning, nematodes were placed in a mixed solution of NaCl and caffeine, and then the chemotaxis index of NaCl was obtained from the nematodes placed on agar medium after pre-exposure to caffeine concentrations of 0.01, 0.1, 0.3, and 1.0%. Locomotor activity and preference behavior for caffeine were also estimated under these caffeine conditions. Nematodes pre-exposed to 0.3% caffeine showed inhibition of salt chemotaxis learning. Additional experiments indicated that nematodes showed a preference response to the middle concentration of caffeine (0.1%), with preference behavior declining in the 0.3% caffeine condition. Stable locomotor activity was observed under 0.01-0.3% caffeine conditions. These results suggest that salt chemotaxis learning with 0.3% caffeine is useful for investigating the effects of caffeine on learning in nematodes.

The Caenorhabditis elegans R13A5.9 gene plays a role in synaptic vesicle exocytosis.

The Caenorhabditis elegans R13A5.9 gene encodes a putative membrane protein with homologs in mammals. When the R13A5.9 protein was fused to different fluorescent proteins, signal was observed in or near synaptic vesicles; thus, we sought to determine whether this gene plays a role in synaptic vesicle formation, function, or exocytosis. R13A5.9 mutant worms exhibited low sensitivity to aldicarb (an acetylcholinesterase inhibitor), which suggested that vesicular loading or release, or acetylcholine synthesis, was disrupted in these organisms. This was supported by the observation that an R13A5.9 mutant strain exhibited an excessive accumulation of synaptic vesicles. Collectively, these results suggest a functional role for R13A5.9 in synaptic vesicle exocytosis.

Navigational choice between reversal and curve during acidic pH avoidance behavior in Caenorhabditis elegans.

BACKGROUND: Under experimental conditions, virtually all behaviors of Caenorhabditis elegans are achieved by combinations of simple locomotion, including forward, reversal movement, turning by deep body bending, and gradual shallow turning. To study how worms regulate these locomotion in response to sensory information, acidic pH avoidance behavior was analyzed by using worm tracking system. RESULTS: In the acidic pH avoidance, we characterized two types of behavioral maneuvers that have similar behavioral sequences in chemotaxis and thermotaxis. A stereotypic reversal-turn-forward sequence of reversal avoidance caused an abrupt random reorientation, and a shallow gradual turn in curve avoidance caused non-random reorientation in a less acidic direction to avoid the acidic pH. Our results suggest that these two maneuvers were each triggered by a distinct threshold pH. A simulation study using the two-distinct-threshold model reproduced the avoidance behavior of the real worm, supporting the presence of the threshold. Threshold pH for both reversal and curve avoidance was altered in mutants with reduced or enhanced glutamatergic signaling from acid-sensing neurons. CONCLUSIONS: C. elegans employ two behavioral maneuvers, reversal (klinokinesis) and curve (klinotaxis) to avoid acidic pH. Unlike the chemotaxis in C. elegans, reversal and curve avoidances were triggered by absolute pH rather than temporal derivative of stimulus concentration in this behavior. The pH threshold is different between reversal and curve avoidance. Mutant studies suggested that the difference results from a differential amount of glutamate released from ASH and ASK chemosensory neurons.

Multiple Chemosensory Neurons Mediate Avoidance Behavior to Rare Earth Ions in Caenorhabditis elegans.

As rare earth (RE) metals are abundantly present in the soil, in spite of their name, it is conceivable that organisms may encounter and interact with RE ions. In the present study, we demonstrated that the soil nematode Caenorhabditis elegans avoids RE ions, such as yttrium and all examined lanthanide ions, which exhibit toxic effects on nematodes. We also demonstrated that the chemosensory system of this animal mediates avoidance behavior toward RE ions similar to heavy metal (HM) ion avoidance. The C. elegans dyf-11(pe554) mutant is unable to respond to chemosensory cues because it lacks all ciliated endings of the chemosensory neurons required for the detection of environmental chemicals. Cell-specific rescue of the dyf-11 mutant and cell-specific genetic ablation studies revealed that the avoidance behavior toward HM and RE ions was mediated by a partially overlapping but distinct subset of chemosensory neurons (ASH, ADL, ASE, ADF, and ASK). With the help of multiple chemosensory neurons, worms may improve the fidelity of avoidance behavior to evade RE ions. Among the chemosensory neurons in C. elegans, ADF and ASK neurons were involved in RE avoidance, but not in HM avoidance. These results suggested that ADF and ASK neurons in C. elegans have RE-selective mechanisms to mediate the avoidance response.

In vivo calcium imaging of OFF-responding ASK chemosensory neurons in C. elegans.

BACKGROUND: How neurons and neuronal circuits transform sensory input into behavior is not well understood. Because of its well-described, simple nervous system, Caenorhabditis elegans is an ideal model organism to study this issue. Transformation of sensory signals into neural activity is a crucial first step in the sensory-motor transformation pathway in an animal's nervous system. We examined the properties of chemosensory ASK neurons of C. elegans during sensory stimulation. METHOD: A genetically encoded calcium sensor protein, G-CaMP, was expressed in ASK neurons of C. elegans, and the intracellular calcium dynamics of the neurons were observed. RESULTS: After application of the attractants l-lysine or food-related stimuli, the level of calcium in ASK neurons decreased. In contrast, responses increased upon stimulus removal. Opposite responses were observed after application and removal of a repellent. CONCLUSION: The observed changes in response to external stimuli suggest that the activity of ASK neurons may impact stimulus-evoked worm behavior. The stimulus-ON/activity-OFF properties of ASK neurons are similar to those of vertebrate retinal photoreceptors. GENERAL SIGNIFICANCE: Analysis of sensory-motor transformation pathways based on the activity and structure of neuronal circuits is an important goal in neurobiology and is practical in C. elegans. Our study provides insights into the mechanism of such transformation in the animal.

Stress Responses Against Rare Earth Ions Are Mediated by the JNK and p38 MAPK Pathways in Caenorhabditis elegans.

Rare earth (RE) ions at high concentrations are toxic to many organisms as they induce oxidative stress and cause improper incorporation of the ions into calcium-binding proteins. Although the mechanism of action underlying the toxicity of REs has been identified, intracellular signaling pathways involved in stress responses against RE ions still remain unclear. In Caenorhabditis elegans, cellular responses against heavy metal stresses are primarily regulated by the c-Jun N-terminal kinase (JNK)-like mitogen-activated protein kinase (MAPK) pathway with a minor contribution of the p38-like MAPK pathway. In this study, we found that both JNK- and p38-like MAPK pathways were involved in stress responses against RE. Unlike heavy metal responses, mutations in both the JNK and p38 pathways caused similar hypersensitivity to RE ions. Although the signaling pathways used for these stress responses were found to be similar, the degree of their respective contribution slightly differed between heavy metal and RE ions.

Modulation of Caenorhabditis elegans chemotaxis by cultivation and assay temperatures.

The chemotaxis behaviors of the nematode Caenorhabditis elegans cultivated at various temperatures (15 degrees C, 20 degrees C and 25 degrees C) were examined at various temperatures (10 degrees C, 15 degrees C, 20 degrees C and 25 degrees C) to determine the multi-sensory integration of physical (thermal) and chemical sensory information within its nervous system. Chemotaxis behavior toward sodium acetate and ammonium chloride were differently affected by both assay and cultivation temperatures, suggesting that the temperature effect on chemotaxis is not general, but rather distinctive for each chemosensory pathway. Since thermosensory cues are likely encountered constantly in C. elegans, we supposed that the chemotaxis behaviors of worms are achieved by the integration of chemo- and thermosensory information. To verify the possible contribution of thermosensory function in chemotaxis, we examined the chemotaxis behaviors of ttx-1(p767) mutant worms with defective AFD thermosensory neurons. The chemotaxis behaviors toward sodium acetate or ammonium chloride of mutant worms cultivated at 20 degrees C and 25 degrees C were reduced relative to those of wild-type worms. These results indicate the important role of multi-sensory integration of chemosensory and thermosensory information in chemotaxis behavior of the C. elegans.

Antibacterial, Antifungal and Nematicidal Activities of Rare Earth Ions.

Despite the name, rare earth elements are relatively abundant in soil. Therefore, these elements might interact with biosphere during the history of life. In this study, we have examined the effect of rare earth ions on the growth of bacteria, fungi and soil nematode. All rare earth ions, except radioactive promethium that we have not tested, showed antibacterial and antifungal activities comparable to that of copper ions, which is widely used as antibacterial metals in our daily life. Rare earth ions also have nematicidal activities as they strongly perturb the embryonic development of the nematode, Caenorhabditis elegans. Interestingly, the nematicidal activity increased with increasing atomic number of lanthanide ions. Since the rare earth ions did not show high toxicity to the human lymphoblastoid cell line or even stimulate the growth of the cultured cells at 1 mM, it raised the possibility that we can substitute rare earth elements for the antibacterial metals usually used because of their safety.

Short-term nicotine exposure induces long-lasting modulation of gustatory plasticity in Caenorhabditis elegans.

Nicotine administration induces many effects on animal behavior. In wild-type Caenorhabditis elegans, gustatory plasticity results in reduced chemotaxis toward NaCl of otherwise attractive concentrations after pre-exposure to 100 mM NaCl in the absence of food. However, acute nicotine administration during a 15 min pre-exposure period inhibits gustatory plasticity, whereas chronic nicotine administration during worm development facilitates the plasticity. To investigate the relationship between the duration of nicotine administration and its effects, we exposed worms to nicotine for various periods during development. The modulatory effect of nicotine on gustatory plasticity was gradually switched from inhibition to facilitation with increased duration of nicotine administration. Moreover, inhibition of plasticity was sustained after relatively short-term chronic administration, with effects lasting for 45 h after the removal of nicotine. Similar to the acute inhibitory effect after 15 min nicotine pre-exposure, the inhibitory effect after short-term chronic administration was dependent on the nicotinic acetylcholine receptor subunit genes lev-1 and unc-29, and genes involved in serotonin biosynthesis bas-1 and tph-1. The impaired inhibition in bas-1 and tph-1mutants was recovered by exogenous serotonin, demonstrating that serotonin plays an important role in the long-lasting inhibitory effects of short-term chronic nicotine exposure.

Food Search Strategy Changes in Caenorhabditis elegans under Chronic Starvation Conditions.

Starvation is a primary threat to survival in nature. This study investigated the effects of starvation on animal behavior and neural function using a nematode model. Nematodes exhibit chemotactic responses to various compounds, including diacetyl produced by food bacteria. Locomotion, chemotactic behavior, and olfactory adaptation were measured following chronic starvation. Our results revealed a starvation-dependent reduction in locomotor activity. Chemotaxis response to the odorant diacetyl was attenuated after 2-38 hr of starvation. However, chemotactic behavior increased significantly after 48 hr of starvation compared with that after 38 hr of starvation, suggesting that food search behavior was enhanced after 48 hr of starvation. Inhibition of diacetyl adaptation was observed in the nematodes after 48 hr of starvation. However, exogenous exposure to serotonin during 48 hr of starvation caused the inhibition of diacetyl adaptation to be attenuated in following 24 hr period of normal feeding.Therefore, the inhibitory effects of starvation on olfactory adaptation may reduce chemotaxis response to the odorant diacetyl in a manner mediated by serotonin.

Neurons regulating the duration of forward locomotion in Caenorhabditis elegans.

The locomotory behavior of Caenorhabditis elegans consists of four simple events, forward and backward movements, omega-shaped turns and rests. The wide variety of behaviors of this worm is achieved through a combination of these simple locomotions. To gain insight into the neuronal mechanisms regulating this locomotion, we analyzed the locomotory behavior of C. elegans over a long time period. By using an automatic worm tracking system, we revealed the existence of at least two distinct behavioral states -- pivoting and traveling -- in the forward locomotion of C. elegans in the absence of food. Pivoting is characterized by pronounced directional switching and resulting in short-duration forward movement, whereas in the traveling state forward movement is of longer duration. Pivoting occurred when we transferred a well-fed worm to an unseeded plate, and then the transition to traveling occurred, successively. We showed that, by laser ablation, antagonistic neuronal pathways consisting of nine classes of sensory neurons and four classes of interneurons were involved in this regulation. Loss of any one of these neurons altered the locomotory behavior.

Effect of simultaneous presentation of multiple attractants on chemotactic response of the nematode Caenorhabditis elegans.

Chemotactic behaviors of the nematode, Caenorhabditis elegans in response to chemical attractants, such as water-soluble sodium acetate and an odorant diacetyl, which were sensed by different sensory neurons, were investigated using various concentrations of these chemical attractants. In the presence of only sodium acetate attractant, the fraction of animals that were roaming around the outside of the attractant and original locations correlated negatively with the chemotaxis index for sodium acetate (P < 0.01). In contrast, the fraction of animals that remained in the original location correlated negatively with the chemotaxis index in the presence of only diacetyl attractant (P < 0.05). These results indicate that the manner of chemotaxis responses differs between sodium acetate and diacetyl. In order to investigate the effect of multiple attractants on chemotactic behaviors, the chemotactic responses to simultaneous presentation of sodium acetate and diacetyl were examined. The fraction of animals that gathered at the 0.7 M sodium acetate location was greater than that at the 0.1% diacetyl location in the presence of both attractants (P < 0.05), although the chemotaxis indexes for 0.7 M sodium acetate and 0.1% diacetyl were similar in the presence of a single attractant. On the other hand, the fraction of animals that gathered at the 0.02% diacetyl location was greater than that at the 0.1M sodium acetate location in the presence of both attractants (P < 0.05), although the chemotaxis indexes for 0.02% diacetyl and 0.1M sodium acetate were similar in the presence of a single attractant. These results suggest the existence of excitatory and/or inhibitory connections in the neuronal circuit for attractant selection, and that the efficacy of these connections may change according to the concentrations of both attractants.

Chemotaxis behavior toward an odor is regulated by constant sodium chloride stimulus in Caenorhabditis elegans.

We studied the chemotaxis behavior of Caenorhabditis elegans toward a chemoattractant in the presence of background sensory stimulus. Chemotaxis toward an odor butanone was greater in the presence of sodium chloride (NaCl) than that without NaCl. By contrast, chemotaxis toward NaCl was not affected by a butanone background. The salt-sensing ASE neuron-deficient che-1(p674) mutants and worms with ASE genetically ablated showed high chemotaxis toward butanone, regardless of the presence of a NaCl background. Therefore, in wild-type worms, information from ASE in the absence of NaCl suppresses butanone chemotaxis, while the suppression is removed in the presence of NaCl.

Odour concentration-dependent olfactory preference change in C. elegans.

The same odorant can induce attractive or repulsive responses depending on its concentration in various animals including humans. However, little is understood about the neuronal basis of this behavioural phenomenon. Here we show that Caenorhabditis elegans avoids high concentrations of odorants that are attractive at low concentrations. Behavioural analyses and computer simulation reveal that the odour concentration-dependent behaviour is primarily generated by klinokinesis, a behavioural strategy in C. elegans. Genetic analyses and lesion experiments show that distinct combinations of sensory neurons function at different concentrations of the odorant; AWC and ASH sensory neurons have critical roles for attraction to or avoidance of the odorant, respectively. Moreover, we found that AWC neurons respond to only lower concentrations of the odorant, whereas ASH neurons respond to only higher concentrations of odorant. Hence, our study suggests that odour concentration coding in C. elegans mostly conforms to the labelled-line principle where distinct neurons respond to distinct stimuli.

Caenorhabditis elegans mutants having altered preference of chemotaxis behavior during simultaneous presentation of two chemoattractants.

Upon presentation of two distinct chemoattractants such as sodium acetate and diacetyl simultaneously, the nematode Caenorhabditis elegans was preferentially attracted by one of these chemoattractants. We isolated two mutants having altered preference of chemotaxis behavior toward simultaneous presentation of sodium acetate and diacetyl. The chep-1(qr1) (CHEmosensory Preference) mutant preferred sodium acetate to diacetyl, while the chep-2(qr2) mutant preferred diacetyl to sodium acetate in simultaneous presentation of these chemoattractants. The chemotaxis behavior of chep-2(qr2) mutant in simultaneous presentation suggests a function of chep-2 gene products within the chemosensory informational integration pathway as well as in the chemosensory pathway.

Serotonin deficiency shortens the duration of forward movement in Caenorhabditis elegans.

Serotonin has been implicated in numerous behaviors in a wide variety of animals. We examined the effect of serotonin deficiency, induced by genetic perturbations and cell ablations, on the duration of Caenorhabditis elegans forward movement. Mutants with defective serotonin biosynthesis or worms with ablated serotonergic neurons showed a markedly decreased duration of forward movement, suggesting involvement of this neuromodulator in the regulation of the duration of worm locomotion.

Chemotaxis of Caenorhabditis elegans during simultaneous presentation of two water-soluble attractants, l-lysine and chloride ions.

Lysine and chloride ions are water-soluble attractants for Caenorhabditis elegans. When chemotaxis behavior to either of these attractants was assayed separately, the radial concentration gradients of 3 M lysine and 0.1 M ammonium chloride had similar potencies for attracting worms. However, when the concentration gradients of lysine and ammonium chloride at these concentrations were presented simultaneously, worms preferred lysine to ammonium chloride more than expected from the results obtained in separate experiments, suggesting the presence of an interaction between these two sensory information pathways within the nervous system. Chemotaxis behavior toward the radial concentration gradient of one of these attractants superimposed on a uniform concentration of the other attractant showed that the chemotaxis was augmented or attenuated by the ammonium chloride background depending on the background concentration, and attenuated by the lysine background, further supporting the interaction between the two sensory information pathways.