Through the looking glass: how do marked dolphins use mirrors and what does it mean?

Mirror-guided self-inspection is seen as a cognitive hallmark purportedly indicating the existence of self-recognition. Only a few species of great apes have been reported to pass a standard mark test for mirror self-recognition in which animals attempt to touch a mark. In addition, evidence for passing the mark test was also reported for Asian elephants, two species of corvids, and a species of cleaner fish. Mirror self-recognition has also been claimed for bottlenose dolphins, using exposure of marked areas to a mirror as evidence. However, what counts as self-directed behaviour to see the mark and what does not has been debated. To avoid this problem, we marked the areas around both eyes of the animals at the same time, one with visible and the other with transparent dye to control for haptic cues. This allowed the animal to see the mark easily and us to investigate what side was exposed to the mirror as an indicator for mark observation. We found that the animals actively chose to inspect their visibly marked side while they did not show an increased interest in a marked conspecific in the pool. These results demonstrate that dolphins use the mirror to inspect their marks and, therefore, likely recognise a distinction between self and others.

Brain monoamine levels and behaviour of young and adult chickens genetically selected on feather pecking.

Severe feather pecking (SFP) in chickens is a detrimental behaviour with possibly neurochemical deficits at its base. Recent neurological studies depicted conflicting results on the role of serotonin (5-hydroxytryptamine, 5-HT) and dopamine (DA) in the development and display of feather pecking. We studied brain monoamine levels and behaviour in domestic chickens divergently genetically selected on feather pecking behaviour, the Low Feather Pecking (LFP) and High Feather Pecking (HFP) lines, both at a young age and when adult, to elucidate the role of 5-HT and DA in feather pecking. Also pecking behaviour and the behavioural response to challenging test situations was determined. At 8 weeks of age, HFP had lower 5-HT and DA turnover in several brain areas than LFP, whereas these differences had disappeared or were even reversed at 25 weeks of age. Line differences in central monoamine activity were found both in emotion-regulating and motor-regulating areas. As expected from previous generations, HFP exceeded LFP in most types of pecking at other birds, including severe feather pecking. Furthermore, HFP responded more actively in most behavioural tests conducted, and seem more impulsive or (hyper)active in their way of coping with challenges. This paper shows different developmental trajectories of the neurochemical systems (5-HT and DA) for chickens divergently selected on feather pecking behaviour, and a remarkable reversion of differences in monoamine activity at a later stage of life. Whether this is a cause or consequence of SFP needs further investigation.

Effects of feather pecking phenotype (severe feather peckers, victims and non-peckers) on serotonergic and dopaminergic activity in four brain areas of laying hens (Gallus gallus domesticus).

Severe feather pecking (SFP) in laying hens is a detrimental behavior causing loss of feathers, skin damage and cannibalism. Previously, we have associated changes in frontal brain serotonin (5-HT) turnover and dopamine (DA) turnover with alterations in feather pecking behavior in young pullets (28-60 days). Here, brain monoamine levels were measured in adult laying hens; focusing on four brain areas that are involved in emotional behavior or are part of the basal ganglia-thalamopallial circuit, which is involved in obsessive compulsive disorders. Three behavioral phenotypes were studied: Severe Feather Peckers (SFPs), Victims of SFP, and Non-Peckers (NPs). Hens (33 weeks old) were sacrificed after a 5-min manual restraint test. SFPs had higher 5-HIAA levels and a higher serotonin turnover (5-HIAA/5-HT) in the dorsal thalamus than NPs, with intermediate levels in victims. NPs had higher 5-HT levels in the medial striatum than victims, with levels of SFPs in between. 5-HT turnover levels did not differ between phenotypes in medial striatum, arcopallium and hippocampus. DA turnover levels were not affected by feather pecking phenotype. These findings indicate that serotonergic neurotransmission in the dorsal thalamus and striatum of adult laying hens depends on differences in behavioral feather pecking phenotype, with, compared to non-pecking hens, changes in both SFP and their victims. Further identification of different SFP phenotypes is needed to elucidate the role of brain monoamines in SFP.

Striatal dopamine D1 receptors are involved in the dissociation of learning based on reward-magnitude.

Here we investigate the contribution of striatal dopamine receptors (D1) to the influence of reward-magnitude on learning. Pigeons (Columba livia) were trained on a discrimination-task with two pairs of stimuli; correct discrimination resulted in a large reward in one pair of stimuli and in a small reward in the other pair. Acquisition of the discrimination-task was accompanied by intracranial injections to the medial striatum, either of a dopamine-antagonist (Sch23390) or of vehicle. In the control-condition the rate of learning was modulated by the magnitude of the reward; discrimination was learned faster if contingent rewards were large and learning was slower if contingent rewards were small. Following injections of D1 antagonist this effect vanished even though the ability to discriminate between the rewards was unaffected. Interestingly, the mean rate of learning was indistinguishable between the control and antagonist conditions. Consequently, it appears that not learning per se but the effect of reward-magnitude on learning is mediated through D1 receptors in the striatum. We argue that the injections of dopamine-antagonist cause a shift in strategy underlying learning. In the control-condition animals rely on positive feedback and thus learning is affected by the magnitude of the contingent reward; in the antagonist-condition, however, learning might rely on negative feedback and is thus insensitive to reward-magnitude.

Quantitative assessment of chronic thalamic stroke.

The procedure presented quantitatively assesses thalamic lesions in the chronic phase of an ischemic episode. The structural MR images of 19 patients with ischemia in the thalamus were assessed by radiologic inspection. An independent rater allocated the damage to the thalamic nuclei. The assessments showed 89% accordance with the radiologic inspection (P < .001). This procedure ranks the extent of the damage to thalamic nuclei and accounts for postacute rearrangement of the neural tissue.

Serotonin 5-HT(1A) receptor binding sites in the brain of the pigeon (Columba livia).

Present knowledge about the serotonergic system in birdbrains is very limited, although the pigeon was used as an animal model in various studies focused on the behavioral effects of serotonergic transmission. In the mammalian brain the 5-HT(1A) receptor is the most widespread serotonin receptor type, and is involved in various functions. Less is known about the distribution of 5-HT(1A) receptors in the avian species. Therefore, we analyzed serotonin 5-HT(1A) receptor binding sites in the pigeon brain using quantitative in vitro receptor autoradiography with the selective radioligand [³H]-8-hydroxy-2-(di-n-propylamino)tetralin ([³H]-8-OH-DPAT). The receptor is differentially distributed throughout the pigeon brain. High levels of 5-HT(1A) receptors are found in the nucleus pretectalis (PT). Moderate densities were detected in the tectum, as well as in the telencephalic nidopallium and hyperpallium. Very low levels were found in the hippocampal formation, the amygdaloid complex, the basal ganglia, and several thalamic nuclei. Furthermore, local variations in 5-HT(1A) receptor densities support the concept of further subdivisions of the entopallium. The regional distribution patterns of 5-HT(1A) receptors mostly display a similar distribution as found in homologue brain structures of mammals.

Telencephalic organization of the olfactory system in homing pigeons (Columba livia).

Pigeons use olfactory cues to navigate over unfamiliar areas, and any impairment of the olfactory system generates remarkable reduction of homing performance. Lesion and deprivation studies suggest a critical involvement of the right nostril and thus, the right olfactory bulb (OB) and the left piriform cortex (CPi) for initial orientation. This functional pattern suggests that OB and CPi are asymmetrically connected with a stronger projection from the right OB to the left CPi. However, the structural organization of the olfactory system is not unequivocally clarified yet. Thus, we re-analyzed the system by antero- and retrograde tract tracing with biotinylated dextran amine and choleratoxin subunit B, and we especially evaluated quantitative differences in the number of cells in the OB innervating the left and right CPi. Our anterograde tracing data verified a strong bilateral input to the CPi, and the prepiriform cortex (CPP), as well as small projections to the ipsilateral medial septum and the dorsolateral corticoid area and the nucleus taeniae of the amygdala in both hemispheres. Apart from the bilateral bulbar afferents, CPi in turn receives unequivocal input from the ipsilateral CPP, hyperpallium densocellulare, dorsal arcopallium, and from a cluster of cells located within the frontolateral nidopallium. Thus, an indirect connection between OB and CPi is only mediated by the CPP. For quantitative analysis of bulbar input to the CPi, we counted the number of ipsi- and contralaterally projecting neurons located in the OB after injections into the left or right CPi. Retrogradely labeled cells were found bilaterally in the OB with a higher number of ipsilaterally located cells. The bilaterality index did not differ after left- or right-sided CPi injections indicating that the functional lateralization of the olfactory system is not simply based on differences in the number of projecting axons of the major processing stream.

The role of dopamine in maintenance and distractability of attention in the "prefrontal cortex" of pigeons.

Selective attention is a crucial component of all sensory processing. Here we test the role of dopamine in attentional selection and in the maintenance of attention. Pigeons were trained on a moving-dot paradigm comparable to the shell game. In this paradigm, pigeons had to select a target among distractors and maintain attention to the target. Target and distractors consisted of white dots, moving at random on a touch-screen. In this task, the demand on attention was modulated by varying the number of distractors and the duration of motion. Both manipulations affected performance equally. In the next step, we investigated the contribution of dopamine to attention. Intracranial injections of D1-antagonist (Sch23390) before testing led to decrements in performance that equally affected trials with different attentional demand. This drop in performance cannot be attributed to altered motivation or motor performance. We conclude that dopamine has a critical role in attention. It is involved in the selection of targets for attention and in the stabilization of attention against interference. This is comparable to the role dopamine plays in working memory and argues for similar mechanisms underlying selective attention and working memory.

Consequences of different housing conditions on brain morphology in laying hens.

The aim of this study was to analyze the impact of physical and social stress on the avian forebrain morphology. Therefore, we used laying hens kept in different housing systems from puberty (approximately 16 weeks old) until the age of 48 weeks: battery cages, small littered ground pen, and free range system. Cell body sizes and catecholaminergic and serotonergic innervation patterns were investigated in brain areas expected to be sensitive to differences in environmental stimulation: hippocampal substructures and the nidopallium caudolaterale (NCL), a functional analogue of the prefrontal cortex. Our analysis shows both structures differing in the affected morphological parameters. Compared to battery cage hens, hens in the free range system developed larger cells in the dorsomedial hippocampus. Only these animals exhibited an asymmetry in the tyrosine hydroxylase density with more fibres in the left dorsomedial hippocampus. We assume that the higher spatial complexity of the free range system is the driving force of these changes. In contrast, in the NCL the housing systems affected only the serotonergic innervation pattern with highest fibre densities in free range hens. Moreover hens of the free range system displayed the worst plumage condition, which most likely is caused by feather pecking causing an altered serotonergic innervation pattern. Considering the remarkable differences between the three housing conditions, their effects on hippocampal structures and the NCL were surprisingly mild. This observation suggests that the adult brain of laying hens displays limited sensitivity to differences in social and physical environment induced post-puberty, which warrants further studies.

Lateralized cognition: asymmetrical and complementary strategies of pigeons during discrimination of the "human concept".

This study was aimed at revealing which cognitive processes are lateralized in visual categorizations of "humans" by pigeons. To this end, pigeons were trained to categorize pictures of humans and then tested binocularly or monocularly (left or right eye) on the learned categorization and for transfer to novel exemplars (Experiment 1). Subsequent tests examined whether they relied on memorized features or on a conceptual strategy, using stimuli composed of new combinations of familiar and novel humans and backgrounds (Experiment 2), whether the hemispheres processed global or local information, using pictures with different levels of scrambling (Experiment 3), and whether they attended to configuration, using distorted human figures (Experiment 4). The results suggest that the left hemisphere employs a category strategy and concentrates on local features, while the right hemisphere uses an exemplar strategy and relies on configuration. These cognitive dichotomies of the cerebral hemispheres are largely shared by humans, suggesting that lateralized cognitive systems already defined the neural architecture of the common ancestor of birds and mammals.

Organization of telencephalotectal projections in pigeons: Impact for lateralized top-down control.

Birds display hemispheric specific modes of visual processing with a dominance of the right eye/left hemisphere for detailed visual object analysis. In pigeons, this behavioral lateralization is accompanied by morphological left-right differences in the ascending tectofugal pathway. This system is also asymmetrically modulated by descending telencephalotectal input whereby the left forebrain displays a much more pronounced physiological control over ipsilateral left and contralateral right visual thalamic processes. In the present study we aimed to answer the question if this top-down asymmetry that up to now had been demonstrated in single cell recording studies is due to anatomical asymmetries in the size of the fiber systems descending from the telencephalon to the tectum. We approached this question by means of a quantitative retrograde tracing study. Cholera toxin subunit B (CtB) was injected unilaterally into either the left or right optic tectum of adult pigeons. After immunohistochemical detection of CtB-positive cells, the number of ipsi- and contralaterally projecting neurons was estimated. Retrogradely labeled cells were located within the arcopallium, the hyperpallium apicale (HA) and the temporo-parieto-occipital area (TPO). Descending projections from HA, arcopallium, and TPO were mainly or exclusively ipsilateral with the contralateral projection being extremely small. Moreover, there was no difference between left and right hemispheric projections. These anatomical data sharply contrast with behavioral and electrophysiological ones which reveal an asymmetric and bilateral top down control. Therefore, contralateral and lateralized forebrain influences onto tectofugal processing are possibly not the direct result of asymmetrical descending axon numbers. Those influences emerge by a lateralized intra- and/or interhemispheric integration of ascending and descending input onto the rotundus.

Transcallosal inhibition across the menstrual cycle: a TMS study.

OBJECTIVE: To determine if there are steroid-dependent changes in transcallosal transfer during the menstrual cycle in normal women. METHODS: We tested 13 normally cycling women during the menstrual, follicular and midluteal phases. Blood levels of estradiol (E) and progesterone (P) were determined by radioimmunoassay. Ipsilateral tonic voluntary muscle activity suppression, called ipsilateral silent period (iSP), was evoked by applying transcranial magnetic stimulation (TMS) over the left motor cortex and by measuring the EMG of the ipsilateral first dorsal interosseus (FDI) muscle. Both iSP-duration and transcallosal conduction times were measured and related to cycle phase and steroid levels. RESULTS: Duration of iSPs varied over the cycle with largest differences between follicular and midluteal phases. During the midluteal phase high levels of P were significantly related to short iSPs. This relation also applied to E levels and iSPs during the follicular phase. CONCLUSIONS: Our study shows for the first time that the transcallosal transfer is modulated by E and P and changes over the menstrual cycle. SIGNIFICANCE: It is suggested that gonadal steroid hormones affect the interhemispheric interaction and change the functional cerebral organization sex dependently via its neuromodulatory properties on GABAergic and glutamatergic neurons.

Neuron numbers in sensory cortices of five delphinids compared to a physeterid, the pygmy sperm whale.

With its large mass and enormous gyrification, the neocortex of whales and dolphins has always been a challenge to neurobiologists. Here we analyse the relationship between neuron number per cortical unit in three different sensory areas and brain mass in six different toothed whale species, five delphinids and one physeterid. Cortex samples, including primary cortical areas of the auditory, visual, and somatosensory systems were taken from both hemispheres of brains fixed in 10% buffered formalin. The samples were embedded in paraffin, sectioned at 25 microm thickness and stained with cresyl violet. Because cortical thickness varies among toothed whale species, cell counts were done in cortical units measuring 150mum in width, 25 microm in thickness, and extending from the pial surface to the white matter. By arranging the delphinid brains according to their total mass, 834-6052 g, we found decreasing neuron numbers in the investigated areas with increasing brain mass. The pigmy sperm whale (Kogia breviceps), a physeterid with an adult brain weight of 1000 g had a distinctly lower neuron number per cortical unit. As had been expected, an increase in adult brain weight in delphinid cetaceans (family Delphinidae) is not correlated with an increase in neuron number per cortical unit.

Right ear advantage for conspecific calls in adults and subadults, but not infants, California sea lions (Zalophus californianus): hemispheric specialization for communication?

This paper explores functional hemispheric asymmetries in the perception of auditory signals in a marine mammal species, the sea lion. Using a head-orienting task toward sounds we found a right ear--left hemisphere--advantage for conspecific calls in adult and subadult California sea lions (Zalophus californianus) that was absent in infants. Non-conspecific sounds did not elicit lateralized reactions in any age group. These findings show that maturational steps regarding communication in the brain of pinnipeds are similar to those described in primates. Such a result in a semi-aquatic species distant from primates on the phylogenetic tree speaks for a stability and an ancient emergence of the left hemispheric specialization for vocal communication. The origin of what seems to be a widespread brain feature might be searched in the temporal and spectral communicative sound's characteristics rather than in its semantic value.

Post-hatch activity-dependent modulation of visual asymmetry formation in pigeons.

The embryonically induced visual lateralization in pigeons can be modified by occlusion of one eye after hatching. Here we show that this deprivation effect could be also attained by short-term blocking of retinal activity with tetrodotoxin (TTX), leading to a dominance of the ipsilateral hemisphere in a visual discrimination task. This lateralization pattern resulted from a performance increase conveyed by the non-deprived hemisphere, while performance with the TTX-injected eye did not differ from that of saline-injected controls. Thus, post-hatch modulation of visual lateralization is mediated by TTX-sensitive, activity-dependent neuronal mechanisms. The transient silencing of one visual input alters the activity balance between the left and right eye system, enhancing visuoperceptive skills in the relatively higher active hemisphere.

Catecholaminergic and dopamine-containing neurons in the spinal cord of pigeons: an immunohistochemical study.

Within the different species belonging to the vertebrate radiation, catecholaminergic elements of the spinal cord present a partly conservative, partly variable pattern. Unfortunately, the overall picture is far from clear since the situation for birds is largely obscure. Therefore, we examined the distribution of dopamine (DA)- and tyrosine hydroxylase (TH)-positive cells and fibers in the spinal cord of the adult pigeon by immunohistochemistry. TH-immunoreactive cells were located within two restricted areas. One group of cells with multipolar shape was located in laminae VI and VII, close to the white-gray border. These cells were more frequently found at rostral and caudal levels while being scarce at cervical-thoracic levels. The second group of cells was located in lamina VIII surrounding the central canal. These cells were bipolar in shape and were found ventrally and laterally to the central canal, with most of them contacting the lumen of the canal through a separate process. The TH-immunoreactive fibers were distributed in both the gray and the white matter. In the gray matter, they were mainly distributed around the central canal (lamina VIII), in the ventral horn close to the border of laminae VII-IX and in the lateral part of the dorsal horn in laminae II-VI. In the white matter the fibers were present in the lateral columns running longitudinal to the main axis. DA-immunoreactive cells were also located within two restricted areas, closely matching the distribution of TH-immunopositive ones. Additionally, the DA-immunoreactive cells had the same shape as the TH-immunoreactive cells, as bipolar neurons contacted the central canal and multipolar ones were located in the laminae VI and VII. Also the distribution of DA- and TH-immunoreactive fibers roughly matched. Both, DA-immunoreactive cells and fibers were scarcer than TH-immunoreactive ones. This finding suggests that the catecholaminergic system in the spinal cord consists of DA-immunoreactive cells as well as other catecholaminergic cells.

Impaired learning of a color reversal task after NMDA receptor blockade in the pigeon (Columba livia) associative forebrain (neostriatum caudolaterale).

The neostriatum caudolaterale (NCL) in the pigeon (Columba livia) forebrain is a multisensory associative area and a functional equivalent to the mammalian prefrontal cortex (PFC). To investigate the role of N-methyl-D-aspartate (NMDA) receptors in the NCL for learning flexibility, the authors trained pigeons in a color reversal task while locally blocking NMDA receptors with D,L-2-2-amino-5-phosphonovalerate (AP-5). Controls received saline injections. AP-5-treated pigeons made significantly more errors and showed significantly stronger perseveration in a learning strategy applied by both groups but were unimpaired in initial learning. Results indicate that NMDA receptors in the NCL are necessary for efficient performance in this PFC-sensitive task, and that they are involved in extinction of obsolete information rather than in acquiring new information.

Electrophysiological and morphological properties of cell types in the chick neostriatum caudolaterale.

The neostriatum caudolaterale, in the chick also referred to as dorsocaudal neostriatal complex, is a polymodal associative area in the forebrain of birds that is involved in sensorimotor integration and memory processes. We have used whole-cell patch-clamp recordings in chick brain slices to characterize the principal cell types of the neostriatum caudolaterale. Electrophysiological properties distinguished four classes of neurons. The morphological characteristics of these classes were examined by intracellular injection of Lucifer Yellow. Type I neurons characteristically fired a brief burst of action potentials. Morphologically, type I neurons had large somata and thick dendrites with many spines. Type II neurons were characterized by a repetitive firing pattern with conspicuous frequency adaptation. Type II neurons also had large somata and thick dendrites with many spines. There was no clear morphological distinction between type I and type II neurons. Type III neurons showed high-frequency firing with little accommodation and a prominent time-dependent inward rectification. They had thin, sparsely spiny dendrites and extensive local axonal arborizations. Electrophysiological and morphological properties indicated them as being interneurons. Type IV neurons had a longer action potential duration, a larger input resistance, and a longer membrane time constant than the other classes. Type IV neurons had small somata and short dendrites with few spines. The long axon collaterals of neurons in all spiny cell classes (types I, II, IV) followed similar patterns, suggesting that neurons from all these types can contribute to the projections of the neostriatum caudolaterale to sensory, limbic and motor areas. The electrophysiological and anatomical characterization of the major classes of neurons in the caudal forebrain of the chick provides a framework for the investigation of sensorimotor integration and learning at the cellular level in birds.

Distribution of BDNF, NT-3, trkB and trkC in the developing retino-tectal system of the pigeon (Columba livia).

The distribution of the neurotrophins BDNF and NT-3 as well as their corresponding high-affinity receptors trkB and trkC was characterized by immunohistochemistry in the developing retino-tectal system of the pigeon. These neurotrophins are known to be important for survival and development of neuronal tissues, but also for activity-dependent neuronal plasticity. In pigeons visual asymmetry is established at the morphological and behavioral level due to a natural asymmetrical light input before hatch, which is followed by a posthatch period of consolidation with unbiased light stimulation. Since the retino-tectal system is the crucial entity of these events, we studied the retinal and the tectal distribution of these neurotrophins and their receptors during retino-tectal formation, to analyze the developmental sequences to which these neurotrophins are tuned. Here we demonstrate that in altricial pigeons no retinal immunolabeling of BDNF, NT-3 or their receptors could be detected before hatch, although a prominent tectal labeling pattern throughout most layers was evident. After hatch, both neurotrophins and their receptors showed a dramatic increase of retinal and tectal distribution. While the tectal and retinal protein synthesis of NT-3 vanished after 2 weeks, that of BDNF could still be revealed in adults. Therefore, the establishment of the retino-tectal system does not seem to depend on these neurotrophins before hatch, although they are probably utilized to shape the intratectal wiring pattern. In contrast, BDNF and NT-3 could play a prominent role in posthatch retino-tectal plasticity, as the consolidation of tectal asymmetries requires posthatch modifications of tectal circuits and proceeds within the first two posthatching weeks. These data are comparable with the distribution of neurotrophins in the retino-tectal system of chicks, although the onset of neurotrophin synthesis seems to be earlier in precocial chicks.