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Ryuzo Shingai

Publications and source records attributed to Ryuzo Shingai.

12 recordsLinked to original sources

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.

Animals↗

Evolution of thermoTRP ion channel homologs in vertebrates.

In mammalian thermosensation, nine temperature-sensitive ion channels that are activated by distinct temperature thresholds have been identified as thermosensors. These ion channels belong to the transient receptor potential (TRP) superfamily and are referred to as "thermoTRPs" (TRPV1, TRPV2, TRPV3, TRPV4, TRPM2, TRPM4, TRPM5, TRPM8, and TRPA1). To elucidate the evolutionary processes of thermoTRPs, we conducted comprehensive searches for mammalian thermoTRP gene homologs in the draft genome sequences of chicken (Gallus gallus), western clawed frog (Xenopus tropicalis), zebrafish (Danio rerio), and pufferfish (Fugu rubripes). Newly identified homologs were compared with known thermoTRPs, and phylogenetic analyses were conducted. Our comparative analyses revealed that most of the mammalian thermo-TRP members already existed in the common ancestor of fishes and tetrapods. Tetrapods shared almost the same repertoire, except that the western clawed frog expanded TRPV4s (six copies) and TRPM8s (two copies), which were diversified considerably. Comparisons of nonsynonymous and synonymous substitution rates among TRPV4s suggested that one copy of the TRPV4 channel in the western clawed frog retained its original function, while the other copies diversified and obtained slightly different properties. In fish lineages, several members of thermo-TRPs have duplicated in the whole genome duplication occurred in the ancestral ray-finned fish; however, some of the copies have subsequently been lost. Furthermore, fishes do not possess the three members of thermoTRPs existed in mammals, e.g., thermoTRPs activated by noxious heat, warm, and cool temperatures. Our results suggest that thermosensation mechanisms have changed through vertebrate evolution with respect to thermosensor repertoires.

Animals↗

Computer-driven automatic identification of locomotion states in Caenorhabditis elegans.

We developed a computer-driven tracking system for the automated analysis of the locomotion of Caenorhabditis elegans. The algorithm for the identification of locomotion states on agar plates (forward movement, backward movement, rest, and curl) includes the identification of the worm's head and tail. The head and tail are first assigned, by using three criteria, based on time-sequential binary images of the worm, and the determination is made based on the majority of the three criteria. By using the majority of the criteria, the robustness was improved. The system allowed us to identify locomotion states and to reconstruct the path of a worm using more than 1h data. Based on 5-min image sequences from a total of 230 individual wild-type worms and 22 mutants, the average error of identification of the head/tail for all strains was 0.20%. The system was used to analyze 70 min of locomotion for wild-type and two mutant strains after a worm was transferred from a seeded plate to a bacteria-free assay plate. The error of identifying the state was less than 1%, which is sufficiently accurate for locomotion studies.

Algorithms↗

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.

Ammonium Chloride↗

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.

Animals↗

Interactions between Caenorhabditis elegans individuals during chemotactic response.

The chemotactic response of the nematode Caenorhabditis elegans is known to be affected by the population density on an assay plate, suggesting the existence of interactions between individual animals. To clarify the interactions between individuals during chemotaxis, we investigated the effect of population density at an attractant area on the chemotactic response to water-soluble sodium acetate and odorant diacetyl using wild-type N2 animals and daf-22 (m130) mutants, which have defective pheromone secretion but can sense pheromone. The chemotaxis index of N2 animals at 90 min of the assay negatively correlated with the number of animals on the assay plate regardless of the type of attractant used (p<0.01). On the other hand, there was no significant difference in the chemotaxis indices of daf-22 (m130) mutants for either of the attractants between the low-and high-population groups. When daf-22 (m130) mutants of a high population density were placed at the attractant location in advance, the chemotaxis index of N2 animals was almost the same as that in the control assay in which no animals were placed at the attractant location in advance. When N2 animals of a high population density were placed at the attractant location in advance, the chemotaxis indices of N2 animals and daf-22 (m130) mutants were significantly smaller than those obtained in the control assay (p<0.05). In the absence of an attractant, we observed a decline in the fraction of animals in the neighborhood of N2 animals of a high population density, although the nematodes were not influenced by daf-22 (m130) mutants of a high population density. These results suggest that the attraction of nematodes to chemicals is inhibited by an increase in the concentration of the pheromone generated by N2 animals at the attractant location.

Animals↗

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.

Animals↗

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.

Animals↗

Neural network model to generate head swing in locomotion of Caenorhabditis elegans.

Computer simulation of the neural network composed of the head neurons of Caenorhabditis elegans was performed to reconstruct the realistic changes in the membrane potential of motoneurons in swinging the head for coordinated forward locomotion. The model neuron had ion channels for calcium and potassium, whose parameters were obtained by fitting the experimental data. Transmission properties of the chemical synapses were set as graded. The neural network involved in forward movement was extracted by tracing the neuronal activity flow upstream from the motoneurons connected to the head muscles. Simulations were performed with datasets, which included all combinations of the excitatory and inhibitory properties of the neurons. In this model, a pulse input entered only from motoneuron VB1, and activation of the stretch receptors on SAA neurons was necessary for the periodic bending. The synaptic output property of each neuron was estimated for the alternate contraction of the dorsal and ventral muscles. The AIB neuron was excitatory, RIV and SMD neurons seemed to be excitatory and RMD and SAA neurons seemed to be inhibitory. With datasets violating Dale's principle for the SMB neuron, AIB neuron was excitatory and RMD neuron was inhibitory. RIA, RIV and SMD neurons seemed to be excitatory.

Animals↗

Inhibition of willardiine-induced currents through rat GluR6/KA-2 kainate receptor channels by Zinc and other divalent cations.

Heteromeric GluR6/KA-2 kainate receptor were expressed in HEK293 cells and an inhibition of willardiine-induced currents by cations was studied. Zinc was much more effective than Ca(2+), Ba(2+) and Mg(2+) at 235, 265 and 1382 fold increase in IC(50), respectively. The inhibition was not voltage-dependent. The present data showed that the binding site for the cations are different from that for willardiine and that the currents are inhibited by the cations via at least two distinct binding sites to Zn(2+) and Ca(2+). These data suggest that Zn(2+) play an important role in modulating glutamate receptors at the nervous system because of a presence of Zn(2+) and various effects of Zn(2+) on the receptors.

Alanine↗

trans-4-Amino-2-methylbut-2-enoic acid (2-MeTACA) and (+/-)-trans-2-aminomethylcyclopropanecarboxylic acid ((+/-)-TAMP) can differentiate rat rho3 from human rho1 and rho2 recombinant GABA(C) receptors.

1. This study investigated the effects of a number of GABA analogues on rat rho3 GABA(C) receptors expressed in Xenopus oocytes using 2-electrode voltage clamp methods. 2. The potency order of agonists was muscimol (EC(50)=1.9 +/- 0.1 microM) (+)-trans-3-aminocyclopentanecarboxylic acids ((+)-TACP; EC(50)=2.7 +/- 0.9 microM) trans-4-aminocrotonic acid (TACA; EC(50)=3.8 +/-0.3 microM) GABA (EC(50)=4.0 +/- 0.3 microM) > thiomuscimol (EC(50)=24.8 +/- 2.6 microM) > (+/-)-cis-2-aminomethylcyclopropane-carboxylic acid ((+/-)-CAMP; EC(50)=52.6 +/-8.7 microM) > cis-4-aminocrotonic acid (CACA; EC(50)=139.4 +/- 5.2 microM). 3. The potency order of antagonists was (+/-)-trans-2-aminomethylcyclopropanecarboxylic acid ((+/-)-TAMP; K(B)=4.8+/-1.8 microM) (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA; K(B)=4.8 +/-0.8 microM) > (piperidin-4-yl)methylphosphinic acid (P4MPA; K(B)=10.2+/-2.3 microM) 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP; K(B)=10.2+/-0.3 microM) imidazole-4-acetic acid (I4AA; K(B)=12.6+/-2.7 microM) > 3-aminopropylphosphonic acid (3-APA; K(B)=35.8+/-13.5 microM). 4. trans-4-Amino-2-methylbut-2-enoic acid (2-MeTACA; 300 microM) had no effect as an agonist or an antagonist indicating that the C2 methyl substituent is sterically interacting with the ligand-binding site of rat rho3 GABA(C) receptors. 5. 2-MeTACA affects rho1 and rho2 but not rho3 GABA(C) receptors. In contrast, (plus minus)-TAMP is a partial agonist at rho1 and rho2 GABA(C) receptors, while at rat rho3 GABA(C) receptors it is an antagonist. Thus, 2-MeTACA and (+/-)-TAMP could be important pharmacological tools because they may functionally differentiate between rho1, rho2 and rho3 GABA(C) receptors in vitro.

Aminobutyrates↗