To identify the neuro-cognitive substrates of valuation and choice, we analysed the neural correlates of anticipated food rewards in the ventral striatum of freely behaving chicks. One-week-old chicks were trained in a color-discrimination task using four color cues (red, yellow, green and blue), each of which was associated with a different food reward. Choosing a red bead was immediately rewarded with a large amount of food, choosing a yellow bead resulted in an immediate-small food reward, and choosing a green bead resulted in a late-large food reward. We selected chicks that consistently chose a large and immediate food reward (red over yellow, and red over green), with the proximity of the food valued higher than the size of the food reward (yellow over green). Of the 47 neurons recorded from the ventral striatum of these chicks, 20 neurons selectively showed cue-period responses to cues associated with food rewards. Five of these 20 neurons responded differentially during the cue period according to the expected delay to reward, and were thus assumed to code for the proximity of the reward. Additionally, three other neurons responded to the quantity of the reward. Furthermore, in the post-operant delay period, many of these 20 neurons showed reward-related activities that were linked to the proximity or presence of the food reward. We therefore propose that impulsive choice and behavioral perseveration observed after lesions of the ventral striatum could be due to impaired anticipation of rewards in the cue and delay periods, respectively.

To reveal the functional roles of the striatum, we examined the effects of excitotoxic lesions to the bilateral medial striatum (mSt) and nucleus accumbens (Ac) in a food reinforcement color discrimination operant task. With a food reward as reinforcement, 1-week-old domestic chicks were trained to peck selectively at red and yellow beads (S+) and not to peck at a blue bead (S−). Those chicks then received either lesions or sham operations and were tested in extinction training sessions, during which yellow turned out to be nonrewarding (S−), whereas red and blue remained unchanged. To further examine the effects on postoperant noninstrumental aspects of behavior, we also measured the “waiting time”, during which chicks stayed at the empty feeder after pecking at yellow. Although the lesioned chicks showed significantly higher error rates in the nonrewarding yellow trials, their postoperant waiting time gradually decreased similarly to the sham controls. Furthermore, the lesioned chicks waited significantly longer than the controls, even from the first extinction block. In the blue trials, both lesioned and sham chicks consistently refrained from pecking, indicating that the delayed extinction was not due to a general disinhibition of pecking. Similarly, no effects were found in the novel training sessions, suggesting that the lesions had selective effects on the extinction of a learned operant. These results suggest that a neural representation of memory-based reward anticipation in the mSt/Ac could contribute to the anticipation error required for extinction.

Effects of rearing conditions were examined in the development of agonistic behaviors in the male fighting fish. In group-I (highly social), fish were communally reared. In group-II (highly social and isolate), fish were individually housed and exposed to the group-I fish through transparent walls until the sexual maturity (from 6 to 12 weeks post-hatch). In group-III (social and isolate), individually housed fish were similarly exposed to other fish within the group. In group-IV (non-social and isolate), individually housed fish were further visually isolated. Agonisitc behaviors were compared among males of the groups-II, -III, and -IV in their fights against the group-I male. The group-IV males showed significantly higher rate of wins than the groups-II and -III males, without differences in the incidence of agonistic behaviors (butt-or-bite, chase, and gill-cover erect) before the termination of the mutual fights. Increased incidence of agonistic behaviors was found after the termination (particularly in the unilateral chase), suggesting that the group-IV males continued to fight even after the opponent male displayed a submission. The aggression was also enhanced in the group-II, when they were thereafter reared in a social isolation after the sexual maturation; a critical period was thus not found. The enhanced aggression was not reversed in the group-IV, when they were thereafter exposed to social stimuli; shift to the continued fights was irreversible. Possible fitness gain of the enhanced aggression was discussed in terms of the adjustability to altered biological resources.

To understand the animal mind, we have to reconstruct how animals recognize the external world through their own eyes. For the reconstruction to be realistic, explanations must be made both in their proximate causes (brain mechanisms) as well as ultimate causes (evolutionary backgrounds). Here, we review recent advances in the behavioral, psychological, and system-neuroscience studies accomplished using the domestic chick as subjects. Diverse behavioral paradigms are compared (such as filial imprinting, sexual imprinting, one-trial passive avoidance learning, and reinforcement operant conditioning) in their behavioral characterizations (development, sensory and motor aspects of functions, fitness gains) and relevant brain mechanisms. We will stress that common brain regions are shared by these distinct paradigms, particularly those in the ventral telencephalic structures such as AIv (in the archistriatum) and LPO (in the medial striatum). Neuronal ensembles in these regions could code the chick's anticipation for forthcoming events, particularly the quality/quantity and the temporal proximity of rewards. Without the internal representation of the anticipated proximity in LPO, behavioral tolerance will be lost, and the chick makes impulsive choice for a less optimized option. Functional roles of these regions proved compatible with their anatomical counterparts in the mammalian brain, thus suggesting that the neural systems linking between the memorized past and the anticipated future have remained highly conservative through the evolution of the amniotic vertebrates during the last 300 million years. With the conservative nature in mind, research efforts should be oriented toward a unifying theory, which could explain behavioral deviations from optimized foraging, such as "naïve curiosity," "contra-freeloading," "Concorde fallacy," and "altruism."

The archistriatum mediates a neural pathway from the medial part of intermediate hyperstriatum ventrale (in the dorsal pallium) to the lobus parolfactorius (in the medial striatum), thus is possibly involved in memory formation in the domestic chick. To elucidate the functional roles, we examined single neuron activities from archistriatum in unconstrained chicks during execution of a GO/NOGO task. In this task, a brief motor sound was given as initial cue, and immediately followed by presentation of a coloured bead. Chick was required to recall the memorized associations between the colour and reward, and pecked at the bead to gain food after a delay (GO trials) or stayed not pecking (NOGO trials). The ventral part of intermediate archistriatum proved to contain a group of neurons that selectively responded to the reward-associated colours before the reward was actually presented, possibly coding the memorized associations. Another group of neurons fired during the reward period, thus could code aspects of the food reward. Yet another group of neurons started to fire immediately on the cue sound and prior to the cued movements nonselectively in both GO and NOGO trials, thus could be involved in the sensori-motor link between the sound and the targeted body movements. It is concluded that even a subregion of archistriatum contains diverse neural codes for memorized associations and food rewards, and neural codes of movements cued by sounds, suggesting that archistriatum is a complex of different functional systems, possibly corresponding to striatum, limbic amygdala, and prefrontal cortex in mammals.

Effects of bilateral chemical lesions of the medial basal ganglia [lobus parolfactorius (LPO)] were examined in 7- to 14-d-old domestic chicks. Chicks were trained in a color discrimination task, in which the subject had to peck one of the two colored beads associated with rewards that differed in quantity (amount of food) and/or temporal proximity (delay of food delivery from peck). In experiment 1, food was given without delay, and chicks successfully learned to choose a colored bead that was associated with a larger reward than the other. In experiment 2, a colored bead (red) was associated with a large reward delivered after a delay (D = 1, 2, or 3 sec), whereas another (yellow) was associated with a small reward delivered immediately. In intact and sham-operated conditions, chicks with a longer D chose the red bead progressively fewer times. Selective lesions to the caudal LPO (but not the rostral LPO) caused impulsive choice, and the ablated chicks chose the yellow bead and gained a small-immediate reward regardless of D. However, when retrained in a null-delay condition (D = 0 sec), the lesioned chick chose the red bead again. Ability to associate novel colors with reward was also unimpaired. These results suggest that the LPO may be responsible for the anticipation of reward proximity and involved in a suppression of impulsiveness by which animals seek immediate gains. The present results also indicate a striking similarity in functional roles between the avian LPO and the nucleus accumbens/ventral striatum in mammals.

To study development of the cerebellum in precocial birds during the early post-hatch period, dendritic arborization of Purkinje neurons was analyzed by intracellular Lucifer-yellow filling in fixed slices. Purkinje neurons were sampled from parasagittal slices of the cerebellar vermis of quail chicks (from 1 day pre-hatch to 14 days post-hatch). Confocal images revealed that the dendritic arborization expanded two-fold during the initial 3 days post-hatch, whereas the growth of cell bodies was much smaller. The dendritic expansion was accompanied by significant elongation of distal dendritic segments (5th and more distal segments), while the length of proximal dendrites (1st-4th segments) and the number of dendritic branches remained unchanged. Active synaptogenesis may occur selectively in distal dendrites during the early post-hatch period.

Newly hatched chicks spontaneously peck at conspicuous objects, and soon learn to discriminate between edible food particles and inedible objects. To examine whether this discrimination is based on a chick's ability to memorize objects by shape cues, we analyzed the pecking behavior. One- to 3-day old quail chicks (Coturnix japonica) were presented with dry objects of different shapes (ball, disk, triangle and T-shape) of similar size (4 mm) and color (green). Habituation occurred after repeated presentation of any one of these objects (duration: 30 sec; interval: 4 min). When chicks showed significantly more pecks at a novel object (dishabituation), we assumed that chicks had memorized the habituated shapes and distinguished the novel object. Chicks did not show dishabituation between a ball and a disk. On the other hand, chicks discriminated a triangle or T-shape from the memorized image of disk, but did not memorize either triangle or T-shape by its shape. Similarly, chicks did not memorize the size of disks as a reference for subsequent pecking behavior. Chicks proved to have a limited ability to memorize shape and size cues for selective pecking behavior, in strong contrast to their accurate memorization of colors.

Effects of bilateral kainate lesions of telencephalic basal ganglia (lobus parolfactorius, LPO) were examined in domestic chicks. In the imprinting paradigm, where chicks learned to selectively approach a moving object without any explicitly associated reward, both the pre- and post-training lesions were without effects. On the other hand, in the water-reinforced pecking task, pre-training lesions of LPO severely impaired immediate reinforcement as well as formation of the association memory. However, post-training LPO lesions did not cause amnesia, and chicks selectively pecked at the reinforced color. The LPO could thus be involved specifically in the evaluation of present rewards and the instantaneous reinforcement of pecking, but not in the execution of selective behavior based on a memorized color cue.

When socially isolated from siblings, newly hatched chicks emit distress calls at a high frequency. As a step toward identifying brain mechanisms responsible for recognition of conspecifics, we analyzed sensory cues critical for suppressing distress calls in socially isolated quail chicks at post-hatch day 2 to 4. Each subject of wild-type chicks was reared with another wild-type companion chick for 1-2 nights before tests. As the first step, we found that distress call was completely suppressed when accompanied by another wild-type chick, irrespective of whether the subject had previously been familiarized with the chick or not. When paired with an albino chick, on the other hand, the subject emitted significantly more calls, suggesting that the wild-type plumage is important. As the next step, in search of the critical sensory modality, we found that visual display of a conspecific chick suppressed the distress call even without accompanying acoustical stimuli. On the other hand, the acoustical stimuli alone were without effects. Olfactory as well as tactile sensations also proved irrelevant. As the final step, we tried to identify the significant sub-modalities in the visual images of conspecifics. Degraded visual image of a moving chick (degraded in fine details) caused a significant but incomplete suppression, whereas an anesthetized chick was without effects. On the other hand, distress calls were completely suppressed even without behavioral correspondence or exact plumage coloration. All of these data suggest that visual image of a moving chick should be given at a high spatial resolution, and wild-type plumage pattern might provide important visual cues.

Properties of local synapses were analyzed in lobus parolfactorius (LPO; avian homologue of caudate-nucleus) of quail chicks by using slice preparations in vitro. Field-potential extracellular and whole-cell intracellular recordings revealed excitatory synaptic inputs converging from dorsal and ventral regions within LPO. With exogenous dopamine (100 μM) in the perfusate, synchronized conditioning stimulation induced biased changes in the dorsal and the ventral inputs; potentiation in the dorsal input and depression in the ventral input in average. On the other hand, de-synchronized conditioning failed to induce such biased changes, although the differences were not statistically significant. SCH-23390 (3 μM) blocked the dorsal potentiation, while AP-5 (100 μM) tended to block both of these changes. The plastic nature may underlie the memory formation in appetitive/aversive learning tasks.

We aimed to reveal what is coded in the basal ganglia of domestic chicks. In the water-reinforced ‘go’ task, chicks learned to peck selectively at a colored bead in order to obtain a drop of water. Out of 38 units obtained, seven showed excitatory activities specifically during the reward period. In the food-reinforced go/no-go task, chicks learned to discriminate two colors to obtain mash food after a delay period. They also learned to ignore another color, which was not associated with a reward. Out of 27 units obtained, four showed excitatory activities during the cue period, specifically when a food reward was anticipated. LPO neurons may code qualities of the obtained rewards, and also chick's anticipation of the forthcoming rewards.

Functional significance of an immediate early gene ZENK (zif/268) was examined in telencephalic regions (homologues of neocortex and basal ganglia) of newly-hatched quail chicks; hyperstriatum accessorium (HA), hyperstriatum ventrale (HV), neostriatum (N) and lobus parolfactorius (LPO). Chicks were trained by a green bead soaked either in a strong aversant (methylanthranilate, MeA), in a weak aversant (MeA diluted by ethanol, 1/3MeA), or in water. Chicks were then tested at 45-50 min post-training, and immediately processed for ZENK immunostaining. Neither the training condition (MeA, 1/3MeA, or water) nor the responses at test (recall or amnesia) significantly contributed to the immunopositive cell densities in all of these regions. On the other hand, single intraperitoneal injection of metrazole (CNS convulsant) induced a transient epileptiform seizure, and caused significantly enhanced ZENK expression in HV and LPO but not in HA and N. However, the metrazol-induced seizure did not interfere with the following passive avoidance training, and chicks successfully learned to avoid the aversive bead when tested at 24 hr subsequently. Among three groups of chicks (metrazol-treated, saline control, and untreated chicks), no significant differences were found in their responses at test (recall, generalized avoidance, or amnesia). These results suggest that enhanced ZENK expression may represent lasting neural activities, but may not be involved specifically in the passive avoidance memory formation.

Newly hatched chicks will spontaneously peck at conspicuous objects in their field of view, and soon learn to distinguish between edible food particles and unpleasant tasting objects. To examine whether the selective pecking is based on the ability to memorize shapes, we analyzed pecking behavior of 1- to 2-days-old quail chicks (Coturnix japonica) by using ball- and triangle-shaped beads both painted in green. Repeated presentation of dry bead (either ball or triangle) resulted in a progressively fewer number of pecks (habituation). When chicks were tested by triangle after repeated presentation of ball, chicks showed a significant increase in the number of pecks at the triangle (dishabituation). On the other hand, when tested by ball after a series of triangle presentations, pecking frequency did not increase (no dishabituation). Chicks thus distinguished the triangle as a novel object after being habituated to ball, but did not respond to the ball after triangle. A similar asymmetry was found in one-trial passive avoidance task. Chicks were pre-trained by water-coated (neutral) triangle and then trained by methylanthranilate-coated (aversive) ball. In this case, most chicks learned to avoid the ball, and half of these successful learners pecked at the triangle; they distinguished triangle from ball. When chicks were pre-trained by neutral ball and trained by aversive triangle, on the other hand, most chicks did not distinguish the ball from triangle, and showed a generalized avoidance for both beads. Chicks may be innately predisposed to memorize a limited category of shapes such as ball, and associate them with selective avoidance.

Animals are predisposed to memorize specific features of objects they encounter, and to link them with behavioral outputs in a selective manner. In this study, we examined whether chicks memorize objects by colors, and how they exploit the memorized color cues for selective pecking in 1- to 2-days-old quail chicks (Coturnix japonica). Ball-shaped beads painted in green (G), yellowish green (YG) and the intermediate color (YGG) were used. Repetitive presentation of a bead (interval: 4.5 min) resulted in gradually fewer pecks (habituation). Subsequent presentation of a different color caused proportionately more pecks (dishabituation); e.g., after habituation to the G bead, the YG bead caused a stronger dishabituation than the YGG bead did. The dishabituation appeared symmetric; e.g., the YG bead caused as strong dishabituation after the G-habituation, as was caused by the G bead after the YG-habituation. Number of pecks could thus reveal the memory-based color perception in chicks. Similar discrimination of beads by memorized color cues was found after one-trial passive avoidance training, where chicks learned to avoid a bitter-tasting object without any differential pre-training experiences. However, proportion of the chicks that discriminated between different colors became progressively smaller at test 15 min, 1 hr, and 24 hr post-training. On the other hand, proportion of chicks that distinguished beads by non-color cues remained unchanged. Chicks may primarily form an accurate memory of colors, but gradually change the link between the color memory and the pecking behavior.

Field potential responses to local electrical stimulation were analyzed in slices of goldfish optic tectum (OT) in vitro. The post-synaptic component of retino–tectal origin was suppressed by DNQX, suggesting glutamatergic field excitatory post-synaptic potential (fEPSP). Repetitive stimulation at 0.1 Hz for 30 min resulted in a slowly developing potentiation in the fEPSP in a population of slices examined. When stimulation at a lower frequency (0.017 Hz for 30 min) preceded the conditioning, the 0.1 Hz stimulation failed to induce the potentiation even though the 0.017 Hz stimulation by itself caused no lasting changes. Induction of the potentiation was partially blocked by AP5 (100 μM), suggesting an involvement of NMDA receptors, however the effect was not significant. We also examined a possible involvement of adenosine, which is supposed to be co-released with glutamate at synaptic terminals. Exogenously applied adenosine (200 μM) slowly depressed fEPSP, and the depression was blocked by DPCPX (A1 receptor antagonist: 400 nM) but not by DMPX (A2 antagonist: 4 μM). However, DPCPX failed to unmask the potentiation after low-frequency stimulation (0.017 Hz for 30 min). Moreover, the potentiability was not correlated with sensitivity to exogenous adenosine. The A1 depression may not underlie the masking effect.

Japanese quails have a variety of calling patterns depending on sex and age. Sexually mature adult males emit a characteristic crow which has a frequency-modulated acoustical trill element. Recently we reported that the intercollicular nucleus of the mesencephalon is the vocal neural system for producing the distress call in untreated chick and the crowing vocalization in testosterone-treated chick in Japanese quails. We postulated that the intercollicular nucleus could mediate this testosterone action on vocal behavior. The present study showed histologically that the intercollicular nucleus neurons of adult males have many more dendrites than those of females. The adult males produced the call with trill element with electrical stimulation of the intercollicular nucleus alone. Females produced the simple call, which has no frequency modulation, with electrical stimulation; 4 days after the testosterone treatment together with electrical stimulation, they also produced the call with trill element, the characteristic call of males. We suggest that the vocal neural system in the intercollicular nucleus is modulated by testosterone during development and produces the crow in males, and the intercollicular nucleus neurons in adults mediate the action of testosterone on vocal behavior. The crow of adult males could therefore be due to effects of testosterone on the neural mechanism in the intercollicular nucleus.

In the intermediate and medial hyperstriatum ventrale (IMHV), a telencephalic region essentially involved in the initial processes of early learning tasks in poultry chicks, induction of an immediate early gene c-fos correlates significantly with the degree of learning (K.V. Anokhin, R. Mileusnic, I.Y. Shamakina, S.P.R. Rose, Effects of early experience on c-fos gene expression in the chick forebrain, Brain Res. 544 (1991) 101-107; B.J. McCabe, G. Horn, Learning-related changes in Fos-like immunoreactivity in the chick forebrain after imprinting, Proc. Natl. Acad. Sci. USA 91 (1994) 11417-11421). In slices of IMHV in vitro, on the other hand, tetanic stimulation at a low frequency induces a potentiation of synaptic responses (P.M. Bradley, B.D. Burns, A.C. Webb, Potentiation of synaptic responses in slices from the chick forebrain, Proc. R. Soc. Lond. B. 243 (1991) 19-24; T. Matsushima, K. Aoki, Potentiation and depotentiation of DNQX-sensitive fast excitatory synaptic transmission in telencephalon of the quail chick, Neurosci. Lett. 185 (1995) 179-182). In this study, we have examined a possible causal link between these two forms of activity-dependent processes, c-fos expression and synaptic potentiation. C-fos was visualized immunohistochemically using antibody raised against the Fos-protein, and potentiation was evaluated on the basis of field potential responses to local electrical stimulation. Tetanic stimulation (5 Hz x 300 pulses) was required for potentiation, but not for c-fos expression. Conversely, a negative correlation appeared between them, and slices with relatively high density of Fos-like immunoreactive cells around the stimulation site failed to show potentiation. Furthermore, drugs similarly effective in blocking potentiation (such as AP5 (NMDA receptor antagonist) and bicuculline (GABA(A) receptor antagonist)) had different effects on the c-fos induction. While AP5 had minor, if any, effects on c-fos expression, bicuculline enhanced it selectively around the site of stimulation. Our results suggest that these two processes are basically distinct, and could represent different aspects in the formation of memory traces in IMHV.