Food-rewarded conditioning and neurophysiological analysis of cheliped gripping behavior in crayfish.

Animals act on their environment to intentionally manipulate it with a defined purpose. This behavior generally needs a special organ suited for the purpose and a highly complex neural mechanism to perform voluntary motor control. Crustaceans with a pair of chelipeds show various manipulative behavior for dietary, exploratory, and reproductive purposes, but the neuronal mechanism underlying the cheliped manipulative behavior has not been clarified yet. In the present study, we trained crayfish Procambarus clarkii to perform a cheliped manipulative task by a newly developed operant paradigm in which animals gripped a specific object for food reward when a visual cue was presented. Animals were then tethered in an operant chamber during the task to enable reliable physiological recordings from the central nervous system. Neural activities descending from the brain were recorded extracellularly from the connective nerves between the brain and the subesophageal ganglion in the trained animals. We found those units showing spike activities that were significantly correlated with cheliped muscle activities, but not with strict timing of visual cue presentation. Although we could not test if those descending activities were necessary or sufficient for initiating the cheliped action by their selective stimulation, the present findings suggest that neural activities for controlling operant gripping behavior are formulated in the brain rather than in the subesophageal ganglion where cheliped motoneurons are present and visual information is transmitted through the brain.

Self-Adjustable Adhesion of Polyampholyte Hydrogels.

Developing nonspecific, fast, and strong adhesives that can glue hydrogels and biotissues substantially promotes the application of hydrogels as biomaterials. Inspired by the ubiquitous adhesiveness of bacteria, it is reported that neutral polyampholyte hydrogels, through their self-adjustable surface, can show rapid, strong, and reversible adhesion to charged hydrogels and biological tissues through the Coulombic interaction.

Electromyographic analysis of goal-directed grasping behavior in the American lobster.

Animals spontaneously initiate goal-directed behavior including foraging action based on their appetitive motivation. The American lobster Homarus americanus exhibits grasping behavior with its crusher claw as feeding behavior that can be initiated after appropriate operant conditioning. In order to quantitatively characterize the goal-directed grasping behavior with a time resolution fine enough for neurophysiological analysis of its initiation and control mechanisms, we made simultaneous electromyographic (EMG) recording from grasping- and reaching-related muscles of the crusher claw while animals initiated grasping behavior. We developed an in vivo extracellular recording chamber that allowed the animal under a semi-restrained condition to perform operant reward learning of claw grasping. Three muscles in the crusher claw (propodite-dactyl closer/opener and coxal protractor) were found to be closely associated with spontaneous grasping behavior. In spontaneous grasping, the activation of those muscles consistently preceded the grasping onset time and exhibited different activity patterns from the grasp induced by a mechanical stimulus. Furthermore, we found that the timing of coxal protractor activation was closer to the grasp onset and its activity was briefer for goal-directed grasping behavior in trained and hungry animals than for non-goal-directed spontaneous grasping behavior in naive or satiated animals. It is suggested that the goal-directed grasping behavior of lobster is characterized, at least partly, by experience-dependent briefer activity of specific muscles involved in reaching action.

Chronic electromyographic analysis of circadian locomotor activity in crayfish.

Animals generally exhibit circadian rhythms of locomotor activity. They initiate locomotor behavior not only reflexively in response to external stimuli but also spontaneously in the absence of any specific stimulus. The neuronal mechanisms underlying circadian locomotor activity can, therefore, be based on the rhythmic changes in either reflexive efficacy or endogenous activity. In crayfish Procambarus clarkii, it can be determined by analyzing electromyographic (EMG) patterns of walking legs whether the walking behavior is initiated reflexively or spontaneously. In this study, we examined quantitatively the leg muscle activity that underlies the locomotor behavior showing circadian rhythms in crayfish. We newly developed a chronic EMG recording system that allowed the animal to freely behave under a tethered condition for more than 10 days. In the LD condition in which the animals exhibited LD entrainment, the rhythmic burst activity of leg muscles for stepping behavior was preceded by non-rhythmic tonic activation that lasted for 1323±488ms when the animal initiated walking. In DD and LL free-running conditions, the pre-burst activation lasted for 1779±31 and 1517±39ms respectively. In the mechanical stimulus-evoked walking, the pre-burst activation ended within 79±6ms. These data suggest that periodic changes in the crayfish locomotor activity under the condition of LD entrainment or free-running are based on activity changes in the spontaneous initiation mechanism of walking behavior rather than those in the sensori-motor pathway connecting mechanoreceptors with leg movements.

Discrimination learning with light stimuli in restrained American lobster.

Operant discrimination learning has been extensively utilized in the study on the perceptual ability of animals and their higher order brain functions. We tested in this study whether American lobster Homarus americanus, which was previously found to possess ability of operant learning with claw gripping, could be trained to discriminate light stimuli of different intensities. For the current purpose, we newly developed a PC-controlled operant chamber that allowed the animal under a body-fixed condition to perform operant reward learning with claw gripping. Lobsters were first reinforced when they gripped the sensor bar upon presentation of a light cue. Then they were trained to grip the bar only when the light stimulus of a specific intensity was presented to obtain food reward while the stimuli of three different intensities including the reinforced one were presented in a random order. Finally, they were re-trained to grip the bar only when the light stimulus of another intensity that was not rewarded in the preceding training to obtain food while other intensities including the one that was rewarded previously were not rewarded any more. In these training procedures, the operant behavior occurred more frequently in response to the rewarded cue than to the non-rewarded one. The action latency for the reinforced stimuli showed a significant decrease in the course of training. These data demonstrate that lobsters can be trained with the light cues of different intensity as discriminative stimuli under a restrained condition that would allow application of electrophysiological techniques to the behaving subjects.

Sequential synaptic excitation and inhibition shape readiness discharge for voluntary behavior.

How do animals initiate voluntary behavior? A key phenomenon in neuroscience is the readiness or preparatory neural activity in specific regions of the animal brain. The neurons and synaptic mechanisms mediating this activity are unknown. We found that the readiness discharge is shaped by sequential synaptic excitation and inhibition in the brain of crayfish (Procambarus clarkii). The readiness discharge neurons extended axon collaterals that appeared to activate recurring local interneurons. Therefore, we propose that the readiness discharge is formed by sequential synaptic events within the brain without feedback signals from downstream ganglia. The circuit involved is suited for signal processing for self-generated voluntary initiation of behavior.

A behavioral analysis of force-controlled operant tasks in American lobster.

Operant conditioning is a common tool for studying cognitive aspects of brain functions. As the first step toward understanding those functions in simple invertebrate microbrains, we tested whether operant conditioning could be applied to train American lobster Homarus americanus that has been extensively adopted as an animal model for neurophysiological analyses of nervous system functions and behavioral control. The animal was trained by food rewarding for gripping of a sensor bar as the operant behavior. Lobsters were first reinforced when they acted on the bar with a stronger grip than a pre-set value. After this reinforcement, the animal learnt to grip the bar for food pellets. The yoked control experiment in which the animal received action-independent reinforcement excluded the possibility of pseudoconditioning that the food simply drove the animal to frequent gripping of the sensor bar. The association of the bar-grip with food was extinguished by rewarding nothing to the operant behavior, and was restored by repeating the reinforcement process as before. In addition, lobsters successfully carried out differential reinforcement regarding the gripping force: their gripping force changed depending on the increased force threshold for food reward. These data demonstrate that lobsters can be trained by operant conditioning paradigms involving acquisition and extinction procedures with the precise claw gripping even under the force control.

Readiness discharge for spontaneous initiation of walking in crayfish.

Animals initiate behavior not only reflexively but also spontaneously in the absence of external stimuli. In vertebrates, electrophysiological data on the neuronal activity associated with the self-initiated voluntary behavior have accumulated extensively. In invertebrates, however, little is known about the neuronal basis of the spontaneous initiation of behavior. We investigated the spike activity of brain neurons at the time of spontaneous initiation of walking in the crayfish Procambarus clarkii and found neuronal signals indicative of readiness or preparatory activities in the vertebrate brain that precede the onset of voluntary actions. Those readiness discharge neurons became active >1 s before the initiation of walking regardless of stepping direction. They remained inactive at the onset of mechanical stimulus-evoked walking in which other descending units were recruited. These results suggest that the parallel descending mechanisms from the brain separately subserve the spontaneous and stimulus-evoked walking. Electrical stimulation of these different classes of neurons caused different types of walking. In addition, we found other descending units that represented different aspects of walking, including those units that showed a sustained activity increase throughout the walking bout depending on its stepping direction, as well as one veto unit for canceling out the output effect of the readiness discharge and three termination units for stopping the walking behavior. These findings suggest that the descending activities are modularized in parallel for spontaneous initiation, continuation, and termination of walking, constituting a sequentially hierarchical control.

Static electric field detection and behavioural avoidance in cockroaches.

Electric fields are pervasively present in the environment and occur both as a result of man-made activities and through natural occurrence. We have analysed the behaviour of cockroaches to static electric fields and determined the physiological mechanisms that underlie their behavioural responses. The behaviour of animals in response to electric fields was tested using a Y-choice chamber with an electric field generated in one arm of the chamber. Locomotory behaviour and avoidance were affected by the magnitude of the electric fields with up to 85% of individuals avoiding the charged arm when the static electric field at the entrance to the arm was above 8-10 kV m(-1). Electric fields were found to cause a deflection of the antennae but when the antennae were surgically ablated, the ability of cockroaches to avoid electric fields was abolished. Fixation of various joints of the antennae indicated that hair plate sensory receptors at the base of the scape were primarily responsible for the detection of electric fields, and when antennal movements about the head-scape joint were prevented cockroaches failed to avoid electric fields. To overcome the technical problem of not being able to carry out electrophysiological analysis in the presence of electric fields, we developed a procedure using magnetic fields combined with the application of iron particles to the antennae to deflect the antennae and analyse the role of thoracic interneurones in signalling this deflection. The avoidance of electric fields in the context of high voltage power lines is discussed.

Electromyographic characterization of walking behavior initiated spontaneously in crayfish.

Crayfish initiate walking behavior not only reflexively in response to external stimuli but also spontaneously in the absence of any specific stimulus. In order to analyze the initiation mechanism underlying these different types of walking, we made simultaneous electromyographic (EMG) recordings from thoracic legs when animals initiated walking, either reflexively or spontaneously, and video recorded their movements synchronously with the EMG recording. Two different stimuli, mechanical and chemical, were used to reflexively induce walking. A non-rhythmic, sustained activation of leg muscles was found to precede the behavioral initiation of either type of walking. The duration of this non-rhythmic muscle activation was significantly longer in the spontaneously initiated walking than in the mechanical stimulus-evoked walking, although no difference was observed between the spontaneous and chemical stimulus-evoked walking. EMG recordings from all eight legs revealed that their non-rhythmic muscle activation occurred almost simultaneously prior to initiation of rhythmical stepping movements. When an animal was suspended without a leg substratum, the timing of muscle activation was more variable among the legs than in the free condition on the substratum. When the circumesophageal commissures were both severed to eliminate signals descending from the brain to the thoracic ganglia, the bilaterally coordinated rhythmic burst activity was not observed in the walking legs. These findings suggest that the spontaneous initiation of walking behavior requires sensory feedback signals from leg proprioceptors, subserved by a different descending activation mechanism from that for stimulus-driven initiation of walking.

Effects of active conductance distribution over dendrites on the synaptic integration in an identified nonspiking interneuron.

The synaptic integration in individual central neuron is critically affected by how active conductances are distributed over dendrites. It has been well known that the dendrites of central neurons are richly endowed with voltage- and ligand-regulated ion conductances. Nonspiking interneurons (NSIs), almost exclusively characteristic to arthropod central nervous systems, do not generate action potentials and hence lack voltage-regulated sodium channels, yet having a variety of voltage-regulated potassium conductances on their dendritic membrane including the one similar to the delayed-rectifier type potassium conductance. It remains unknown, however, how the active conductances are distributed over dendrites and how the synaptic integration is affected by those conductances in NSIs and other invertebrate neurons where the cell body is not included in the signal pathway from input synapses to output sites. In the present study, we quantitatively investigated the functional significance of active conductance distribution pattern in the spatio-temporal spread of synaptic potentials over dendrites of an identified NSI in the crayfish central nervous system by computer simulation. We systematically changed the distribution pattern of active conductances in the neuron's multicompartment model and examined how the synaptic potential waveform was affected by each distribution pattern. It was revealed that specific patterns of nonuniform distribution of potassium conductances were consistent, while other patterns were not, with the waveform of compound synaptic potentials recorded physiologically in the major input-output pathway of the cell, suggesting that the possibility of nonuniform distribution of potassium conductances over the dendrite cannot be excluded as well as the possibility of uniform distribution. Local synaptic circuits involving input and output synapses on the same branch or on the same side were found to be potentially affected under the condition of nonuniform distribution while operation of the major input-output pathway from the soma side to the one on the opposite side remained the same under both conditions of uniform and nonuniform distribution of potassium conductances over the NSI dendrite.

Disynaptic and polysynaptic statocyst pathways to an identified set of premotor nonspiking interneurons in the crayfish brain.

Nonspiking giant interneurons (NGIs) in the crayfish brain occupy a key position in the neuronal circuit for eyestalk motor control and, hence, play a crucial role in the central compensation process following unilateral deprivation of the statocyst that functions as an equilibrium sensory system. The synaptic organization of their input pathways, however, remains unknown. In the present study we identified 11 local interneurons that were polysynaptically connected to NGIs by making simultaneous intracellular recordings. We also identified three other local interneurons that connected to NGIs monosynaptically. PLNI-2 was a nonspiking interneuron making an excitatory synaptic connection to an NGI that had its cell body on the opposite side. PLSI was a spiking interneuron that made an inhibitory connection to an ipsilateral NGI. These cells were entirely confined to the protocerebrum. Another local spiking interneuron termed UGLI-1 was found to make an excitatory connection with a contralateral NGI, extending dendrites in the anterior and posterior medial protocerebral neuropils and the lateral antenna I neuropil in the deutocerebrum where statocyst afferents project. When a depolarizing current was injected into the UGLI-1, the frequency of discrete excitatory PSPs increased remarkably in the postsynaptic NGI, each PSP following the UGLI-1 spike in one-to-one correspondence. The UGLI was previously reported to be activated monosynaptically by statocyst afferents. The present study thus finally demonstrates the tri-synaptic organization of the statocyst-to-eyestalk motor neuron pathway in its simplest form, suggesting the critical role of the UGLI-1 in the central compensation.

Gaseous neuromodulator-related genes expressed in the brain of honeybee Apis mellifera.

Nitric oxide (NO), hydrogen sulfide (H2S), and carbon monoxide (CO) are thought to act as gaseous neuromodulators in the brain across species. For example, in the brain of honeybee Apis mellifera, NO plays important roles in olfactory learning and discrimination, but the existence of H2S- and CO-mediated signaling pathways remains unknown. In the present study, we identified the genes of nitric oxide synthase (NOS), soluble guanylyl cyclase (sGC), cystathionine beta-synthase (CBS), and heme oxygenase (HO) from the honeybee brain. The honeybee brain contains at least one gene for each of NOS, CBS, and HO. The deduced proteins for NOS, CBS, and HO are thought to contain domains to generate NO, H2S, and CO, respectively, and to contain putative Ca2+/calmodulin-binding domains. On the other hand, the honeybee brain contains three subunits of sGC: sGCalpha1, sGCbeta1, and sGCbeta3. Phylogenetic analysis of sGC revealed that Apis sGCalpha1 and sGCbeta1 are closely related to NO- and CO-sensitive sGC subunits, whereas Apis sGCbeta3 is closely related to insect O2-sensitive sGC subunits. In addition, we performed in situ hybridization for Apis NOS mRNA and NADPH-diaphorase histochemistry in the honeybee brain. The NOS gene was strongly expressed in the optic lobes and in the Kenyon cells of the mushroom bodies. NOS activity was detected in the optic lobes, the mushroom bodies, the central body complex, the lateral protocerebral lobes, and the antennal lobes. These findings suggest that NO is involved in various brain functions and that H2S and CO can be endogenously produced in the honeybee brain.

Physiological changes of premotor nonspiking interneurons in the central compensation of eyestalk posture following unilateral sensory ablation in crayfish.

We investigated how the physiological characteristics and synaptic activities of nonspiking giant interneurons (NGIs), which integrate sensory inputs in the brain and send synaptic outputs to oculomotor neurons innervating eyestalk muscles, changed after unilateral ablation of the statocyst in order to clarify neuronal mechanisms underlying the central compensation process in crayfish. The input resistance and membrane time constant in recovered animals that restored the original symmetrical eyestalk posture 2 weeks after operation were significantly greater than those immediately after operation on the operated side whereas in non-recovered animals only the membrane time constant showed a significant increase. On the intact side, both recovered and non-recovered animals showed no difference. The frequency of synaptic activity showed a complex pattern of change on both sides depending on the polarity of the synaptic potential. The synaptic activity returned to the bilaterally symmetrical level in recovered animals while bilateral asymmetry remained in non-recovered ones. These results suggest that the central compensation of eyestalk posture following unilateral impairment of the statocyst is subserved by not only changes in the physiological characteristics of the NGI membrane but also the activity of neuronal circuits presynaptic to NGIs.

Functional significance of passive and active dendritic properties in the synaptic integration by an identified nonspiking interneuron of crayfish.

Nonspiking interneurons control their synaptic output directly by membrane potential changes caused by synaptic activities. Although these interneurons do not generate spikes, their dendritic membrane is endowed with a variety of voltage-dependent conductances whose functional significance in synaptic integration remains unknown. We quantitatively investigated how the passive and active dendritic properties affect the synaptic integration in an identified nonspiking interneuron of crayfish by computer simulation using its multicompartment model based on electrophysiological measurements and three-dimensional morphometry. At the resting potential level, the attenuation factor (V(s)/V(t)) of a unitary synaptic potential in the course of its spread from a dendritic terminal (V(s)) to other terminals (V(t)) ranged from 4.42 to 6.30 with no substantial difference between hyperpolarizing and depolarizing potentials. The compound synaptic responses to strong mechanosensory stimulation could be reproduced in calculation only as the result of spatial summation of attenuated potentials, not as any single large potential. The characteristic response could be reproduced by assuming that the active conductances were distributed only in the dendritic region where the synaptic summation was carried out. The active conductances in other parts of the cell affected neither the shape of the compound synaptic response nor the dendritic spread of synaptic potentials. These findings suggest that the active membrane conductances do not affect the spatial distribution of synaptic potentials over dendrites but function in sculpting the summed synaptic potential to enhance temporal resolution in the synaptic output of the nonspiking interneuron.

Further exploration into the adaptive design of the arthropod "microbrain": I. Sensory and memory-processing systems.

Arthropods have small but sophisticated brains that have enabled them to adapt their behavior to a diverse range of environments. In this review, we first discuss some of general characteristics of the arthropod "microbrain" in comparison with the mammalian "megalobrain". Then we discuss about recent progress in the study of sensory and memory-processing systems of the arthropod "microbrain". Results of recent studies have shown that (1) insects have excellent capability for elemental and context-dependent forms of olfactory learning, (2) mushroom bodies, higher olfactory and associative centers of arthropods, have much more elaborated internal structures than previously thought, (3) many genes involved in the formation of basic brain structures are common among arthropods and vertebrates, suggesting that common ancestors of arthropods and vertebrates already had organized head ganglia, and (4) the basic organization of sensori-motor pathways of the insect brain has features common to that of the mammalian brain. These findings provide a starting point for the study of brain mechanisms of elaborated behaviors of arthropods, many of which remain unexplored.

Characterization of electro-olfactogram oscillations and their computational reconstruction.

Electro-olfactogram (EOG) oscillations induced by odorant stimulation have been often reported in various vertebrates from fishes to mammals. However, the mechanism of generation of EOG oscillations remains unclear. In the present study, we first characterized the properties of EOG oscillations induced by amino acid odorants in the rainbow trout and then performed a computer simulation based on the main assumption that olfactory receptor neurons (ORNs) have intrinsic oscillatory properties due to two types of voltage-gated ion channels, which have not yet been reported in vertebrate ORNs. EOG oscillations appeared mostly on the peak and decay phases of negative EOG responses, when odorant stimuli at high intensity flowed regularly anterior to posterior olfactory lamellae in the olfactory organ. The appearance of EOG oscillations was dependent on the odorant intensity but not on the flow rate. The maximum amplitude and the maximum power frequency of EOG oscillations were 3.51 +/- 3.35 mV (mean +/- SD, n = 232, range 0.12-16.79 mV) and 10.59 +/- 5.05 Hz (mean +/- SD, n = 232, range 3.51-40.03 Hz), respectively. The simulation represented sufficiently well the characteristics of EOG oscillations; occurrence at high odorant concentration, odorant concentration-dependent amplitude and the maximum power frequency range actually observed. Our results suggest that EOG oscillations are due to the intrinsic oscillatory properties of individual ORNs, which have two novel types of voltage-gated ion channels (resonant and amplifying channels). The simulation program for Macintosh ('oscillation 3.2.4' for MacOS 8.6 or later) is available on the world wide web (http://bio2.sci.hokudai.ac.jp/bio/chinou1/noriyo_home.html).

An optical telemetry system for underwater recording of electromyogram and neuronal activity from non-tethered crayfish.

We have developed an optical telemetry system for recording electrical signals associated with muscle and neuronal activities from freely walking crayfish under water. The device was made from conventional electronic parts which are commercially available, utilizing infrared light (880 nm) for signal transmission. Two or four channels of biological signals were multiplexed, the voltage of each data point modulated to the duration of subcarrier pulses and further to the interval of narrower carrier pulses that directly drove the infrared light emission diode (IRLED) under water. The light-pulse modulated signals were received by photodiodes and demodulated to restore the original two or four channel signals. Electrical recordings using wired electrodes and conventional amplifiers revealed that the optically transmitted signals were consistent with the wire-transmitted ones. In order to test the performance of this system, we recorded electromyograms (EMGs) from the second and third walking legs on each side of crayfish together with the neuronal activity in the ventral nerve cord. The results confirmed our previous observation in tethered crayfish that the background tonus of leg muscles showed an increase preceding their rhythmic activation.

Mechanoreceptors involved in the hindwing-evoked escape behaviour in cricket, Gryllus bimaculatus.

Mechanoreceptors involved in the escape jumping evoked by hindwing stimulation have been investigated in the field cricket Gryllus bimaculatus. By partial ablation of the hindwing, we found that a mechanosensory system relevant to the escape behaviour was localized on specific veins of the hindwing tip. Scanning electron microscopy revealed three types of mechanoreceptive sensillae on the corresponding region. Based on their morphology, type I and type III sensillae were judged to be trichoid and chaetic sensillae, respectively. Type II sensillae were newly found in this study, having a twisted shaft with a socket-like structure at its base. They existed almost exclusively on the tip and middle regions of the hindwing. The conduction velocity of type II units was significantly smaller than that of type I units. One cycle of sinusoidal deflection of a single type II sensilla at frequencies in the range of 10-120 Hz caused the sensory unit to discharge a single or a few spikes that were not directly correlated with any specific direction of hair movement nor specific deflection angle. The response probability decreased with the stimulus frequency to be less than 0.1 at 0.2 Hz. The results suggest that type II sensillae would serve as contact mechanoreceptors with a low-cut filter property to obtain general information on the presence of stimuli on the hindwing tip rather than specific information on their precise positioning or movement.

Oscillatory current responses of olfactory receptor neurons to odorants and computer simulation based on a cyclic AMP transduction model.

Neural oscillatory activities triggered by odorant stimulation have been often reported at various levels of olfactory nervous systems in vertebrates. To elucidate the origin of neural oscillations, we studied first the oscillatory properties of current responses of isolated olfactory receptor neurons (ORNs) of the rainbow trout to amino acid odorants, using a whole-cell voltage-clamp technique and found that the damped current oscillations were intrinsic in both ciliated and microvillous ORNs and occurred when ORNs were stimulated by odorants at high intensities. Continuous wavelet analysis using the Gabor function revealed that the dominant frequency of oscillations was 1.89 +/- 0.50 Hz (mean +/- SD, n = 92). There was no significant difference in oscillation frequency between the two types of ORNs and between different perfusion conditions with standard and Na(+)-free (choline) Ringer's solutions, but there was a slight difference in oscillation frequency between different holding potential conditions of negative and positive potentials. We then performed a computer simulation of the current responses with a cAMP olfactory transduction model. The model was based on the assumption that the current responses of ORNs were linearly related to the sum of concentrations of active cyclic-nucleotide-gated channels and Ca(2+)-activated Cl(-) channels, and was expressed by 12 differential equations with 44 different parameters. The simulation revealed that the oscillations of current responses of ORNs were mainly due to the oscillatory properties of intracellular cAMP and Ca(2+) concentrations. The necessary reaction component for the oscillations in the transduction model was direct inhibition of adenylate cyclase activity by Ca(2+). High Ca(2+) efflux by the Na(+)-Ca(2+) exchanger and cAMP-phosphodiesterase activity were most influential on the oscillations. The simulation completely represented the characteristics of current responses of ORNs: odorant-intensity-dependent response, intensity-dependent latency and adaptation. Thus, the simulation is generally applicable to current and voltage responses of ORNs equipped with cAMP olfactory transduction pathway in other vertebrate species. The simulation programs for Macintosh (cAMP 9.2.7 and 9.2.8 for MacOS 8.1 or later) and cAMP JAVA applet versions based on cAMP 9.2.8 have been published on the world wide web (http://bio2.sci.hokudai.ac.jp/bio/chinou1/noriyo_home.html).