Origin and evolution of human cognition.

We investigate which of the higher cognitive abilities or types of intelligence characteristic of humans are found, even in preliminary form, in non-human animals, predominantly primates, or whether qualitatively different ("unique") human abilities exist. This concerns (1) tool use and fabrication, (2) problem solving, (3) gaze following, (4) mirror self-recognition, (5) imitation, (6) metacognition, (7) theory of mind, (8) consciousness, (9) prosociality, and (10) language. We found that none of these abilities can be regarded as unique to humans without precursors in non-human primates. The observed differences in cognitive functions, underlying brain mechanisms and resulting behaviors correlate best with differences in the information processing capacity as an equivalent of general intelligence based on the number of cortical neurons, packing density and axonal conduction velocity plus long-range cortical fascicles. The biggest quantitative change appears to concern the origin of syntactical language, but this was preceded by an increased mental ability to manipulate sequential events within working memory.

Chemical characterization of the adhesive secretions of the salamander Plethodon shermani (Caudata, Plethodontidae).

Salamanders have developed a wide variety of antipredator mechanisms, including tail autotomy, colour patterns, and noxious skin secretions. As an addition to these tactics, the red-legged salamander (Plethodon shermani) uses adhesive secretions as part of its defensive strategy. The high bonding strength, the fast-curing nature, and the composition of the biobased materials makes salamander adhesives interesting for practical applications in the medical sector. To understand the adhesive secretions of P. shermani, its components were chemically analysed by energy dispersive X-ray spectroscopy (EDX), inductively coupled plasma mass spectrometry (ICP-MS), amino acid analysis, and spectroscopy (ATR-IR, Raman). In addition, proteins were separated by gel-electrophoresis and selected spots were characterised by peptide mass fingerprinting. The salamander secretion contains a high amount of water and predominantly proteins (around 77% in the dry stage). The gel-electrophoresis and peptide mass fingerprint analyses revealed a de novo set of peptides/proteins, largely with a pI between 5.0 and 8.0 and a molecular mass distribution between 10 and 170 kDa. Only low homologies with other proteins present in known databases could be identified. The results indicate that the secretions of the salamander Plethodon clearly differ chemically from those shown for other glue-producing terrestrial or marine species and thus represent a unique glue system.

Salamanders on the bench - A biocompatibility study of salamander skin secretions in cell cultures.

Salamanders have evolved a wide variety of antipredator mechanisms and behavior patterns, including toxins and noxious or adhesive skin secretions. The high bonding strength of the natural bioadhesives makes these substances interesting for biomimetic research and applications in industrial and medical sectors. Secretions of toxic species may help to understand the direct effect of harmful substances on the cellular level. In the present study, the biocompatibility of adhesive secretions from four salamander species (Plethodon shermani, Plethodon glutinosus, Ambystoma maculatum, Ambystoma opacum) were analyzed using the MTT assay in cell culture and evaluated against toxic secretions of Pleurodeles waltl, Triturus carnifex, Pseudotriton ruber, Tylototriton verrucosus, and Salamandra salamandra. Their effect on cells was tested in direct contact (direct culture) or under the influence of the extract (indirect exposure) in accordance with the protocol of the international standard norm ISO 10993-5. Human dermal fibroblasts (NHDF), umbilical vein endothelial cells (HUVEC), and articular chondrocytes (HAC), as well as the cell lines C2C12 and L929 were used in both culture types. While the adhesive secretions from Plethodon shermani are cytocompatible and those of Ambystoma opacum are even advantageous, those of Plethodon glutinosus and Ambystoma maculatum appear to be cytotoxic to NDHF and HUVEC. Toxic secretions from Salamandra salamandra exhibited harmful effects on all cell types. Pseudotriton ruber and Triturus carnifex secretions affected certain cell types marginally; those from Pleurodeles waltl and Tylototriton verrucosus were generally well tolerated. The study shows for the first time the effect of salamander secretions on the viability of different cell types in culture. Two adhesive secretions appeared to be cell compatible and are therefore promising candidates for future investigations in the field of medical bioadhesives. Among the toxic secretions tested, only two of the five had a harmful effect on cells, indicating different cell toxicity mechanisms.

Lesions of the dorsal striatum impair orienting behaviour of salamanders without affecting visual processing in the tectum.

In amphibians, visual information in the midbrain tectum is relayed via the thalamus to telencephalic centres. Lesions of the dorsal thalamus of the salamander Plethodon shermani result in impairment of orienting behaviour and in modulation of spike pattern of tectal neurons. These effects may be induced by an interruption of a tectum-thalamus-telencephalon-tectum feedback loop enabling spatial attention and selection of visual objects. The striatum is a potential candidate for involvement in this pathway; accordingly, we investigated the effects of lesioning the dorsal striatum. Compared to controls and sham lesioned salamanders, striatum-lesioned animals exhibited a significantly lower number of orienting responses toward one of two competing prey stimuli. Orienting towards stimuli was impaired, while the spike pattern of tectal cells was unaffected, because both in controls and striatum-lesioned salamanders the spike number significantly decreased at presentation of one prey stimulus inside the excitatory receptive field and another one in the surround compared to that at single presentation inside the excitatory receptive field. We conclude that the dorsal striatum contributes to orienting behaviour, but not to an inhibitory feedback signal onto tectal neurons. The brain area engaged in the feedback loop during visual object discrimination and selection has yet to be identified. Information processing in the amphibian striatum includes multisensory integration; the striatum generates behavioural patterns that influence (pre)motor processing in the brainstem. This situation resembles the situation found in rats, in which the dorsolateral striatum is involved in stimulus-response learning regardless of the sensory modality, as well as in habit formation.

Neuronal factors determining high intelligence.

Many attempts have been made to correlate degrees of both animal and human intelligence with brain properties. With respect to mammals, a much-discussed trait concerns absolute and relative brain size, either uncorrected or corrected for body size. However, the correlation of both with degrees of intelligence yields large inconsistencies, because although they are regarded as the most intelligent mammals, monkeys and apes, including humans, have neither the absolutely nor the relatively largest brains. The best fit between brain traits and degrees of intelligence among mammals is reached by a combination of the number of cortical neurons, neuron packing density, interneuronal distance and axonal conduction velocity--factors that determine general information processing capacity (IPC), as reflected by general intelligence. The highest IPC is found in humans, followed by the great apes, Old World and New World monkeys. The IPC of cetaceans and elephants is much lower because of a thin cortex, low neuron packing density and low axonal conduction velocity. By contrast, corvid and psittacid birds have very small and densely packed pallial neurons and relatively many neurons, which, despite very small brain volumes, might explain their high intelligence. The evolution of a syntactical and grammatical language in humans most probably has served as an additional intelligence amplifier, which may have happened in songbirds and psittacids in a convergent manner.

Morphological characterization of the glandular system in the salamander Plethodon shermani (Caudata, Plethodontidae).

Amphibians have evolved a wide variety of mechanisms that provide a certain degree of protection against predators, including camouflage, tail autonomy, encounter behavior and noxious or toxic skin secretions. In addition to these strategies, some amphibians release a glue-like secretion onto the surface of their skin when threatened. While some information regarding the origin and production of these adhesive secretions is available for frogs such as Notaden bennetti, these aspects are only partially understood in salamanders. We contribute to an earlier study and provide additional information regarding the origin, production, and characterization of the adhesive secretion in the red-legged salamander (Plethodon shermani) at a microanatomical level. When stressed, this salamander secretes a milky, viscous liquid from its dorsal and ventral skin. This secretion is extremely adhesive and hardens within seconds upon exposure to air. This study describes two cutaneous gland types (mucous and granular) in the dorsal and ventral epithelial tissue that differ considerably in their secretory content. While the smaller mucous glands contains flocculent to granular material, mostly acidic glycoproteins, the granular glands synthesize various granules of differing size and density that consist of basic proteinaceous material. The results strongly indicate that the secretions of both gland types from the dorsal as well as the ventral side form the adhesive mucus in Plethodon shermani, consisting of basic and acidic glycoproteins, glycoconjugates with mannose and α-L-fucose residues as well as lipid components.

The role of the dorsal thalamus in visual processing and object selection: a case of an attentional system in amphibians.

In amphibians, the midbrain tectum is regarded as the visual centre for object recognition but the functional role of forebrain centres in visual information processing is less clear. In order to address this question, the dorsal thalamus was lesioned in the salamander Plethodon shermani, and the effects on orienting behaviour or on visual processing in the tectum were investigated. In a two-alternative-choice task, the average number of orienting responses toward one of two competing prey or simple configural stimuli was significantly decreased in lesioned animals compared to that of controls and sham-lesioned animals. When stimuli were presented during recording from tectal neurons, the number of spikes on presentation of a stimulus in the excitatory receptive field and a second salient stimulus in the surround was significantly reduced in controls and sham-lesioned salamanders compared to single presentation of the stimulus in the excitatory receptive field, while this inhibitory effect on the number of spikes of tectal neurons was absent in thalamus-lesioned animals. In amphibians, the dorsal thalamus is part of the second visual pathway which extends from the tectum via the thalamus to the telencephalon. A feedback loop to the tectum is assumed to modulate visual processing in the tectum and to ensure orienting behaviour toward visual objects. It is concluded that the tectum-thalamus-telencephalon pathway contributes to the recognition and evaluation of objects and enables spatial attention in object selection. This attentional system in amphibians resembles that found in mammals and illustrates the essential role of attention for goal-directed visuomotor action.

Evolution of the brain and intelligence in primates.

Primates are, on average, more intelligent than other mammals, with great apes and finally humans on top. They generally have larger brains and cortices, and because of higher relative cortex volume and neuron packing density (NPD), they have much more cortical neurons than other mammalian taxa with the same brain size. Likewise, information processing capacity is generally higher in primates due to short interneuronal distance and high axonal conduction velocity. Across primate taxa, differences in intelligence correlate best with differences in number of cortical neurons and synapses plus information processing speed. The human brain stands out by having a large cortical volume with relatively high NPD, high conduction velocity, and high cortical parcellation. All aspects of human intelligence are present at least in rudimentary form in nonhuman primates or some mammals or vertebrates except syntactical language. The latter can be regarded as a very potent "intelligence amplifier."

Quantity discrimination in salamanders.

We investigated discrimination of large quantities in salamanders of the genus Plethodon. Animals were challenged with two different quantities (8 vs 12 or 8 vs 16) in a two-alternative choice task. Stimuli were live crickets, videos of live crickets or images animated by a computer program. Salamanders reliably chose the larger of two quantities when the ratio between the sets was 1:2 and stimuli were live crickets or videos thereof. Magnitude discrimination was not successful when the ratio was 2:3, or when the ratio was 1:2 when stimuli were computer animated. Analysis of the salamanders' success and failure as well as analysis of stimulus features points towards movement as a dominant feature for quantity discrimination. The results are generally consistent with large quantity discrimination investigated in many other animals (e.g. primates, fish), current models of quantity representation (analogue magnitudes) and data on sensory aspects of amphibian prey-catching behaviour (neuronal motion processing).

Allografting combined with systemic FK506 produces greater functional recovery than conduit implantation in a rat model of sciatic nerve injury.

Implantation of allografts or nerve conduits has been used to promote regeneration following peripheral nerve injuries involving substantial axon loss. Both methods provide promising alternatives to autologous grafting and avoid donor site morbidity. We compared the relative efficacies of allografting versus conduit implantation in a rat model of sciatic nerve regeneration. Two rat strains (Lewis and Dark Agouti; n = 30) were employed. Unoperated animals served as controls (group I). Animals in groups II and III underwent left sciatic nerve resection over a distance of 15 mm; group II animals received implants of collagen type I conduits; and group III animals received allografts from the other rat strain and systemic low-dose (0.1 mg/kg/d) administration of FK506. Walking tracks were recorded after 4, 8, 12, and 16 weeks; nerve sections were stained for myelin basic protein after 16 weeks. Functional tests revealed significantly better recovery in group III animals compared with group II even though there was no significant difference in the extent of remyelination. Neither group achieved the functional or histomorphometric values of control animals. Improved functional recovery following allografting plus systemic FK506, in comparison with conduit implantation, underlines the importance of systemic administration of neurotrophic molecules for nerve regeneration.

Administration of low-dose FK 506 accelerates histomorphometric regeneration and functional outcomes after allograft nerve repair in a rat model.

A substantial loss of peripheral nerves requires grafts for repair. In animal experiments, the use of allografts is successful only when rejection of the transplant is prevented and nerve regeneration is improved by the administration of the immunosuppressant FK 506 used in high doses. In this study, we examined the functional and morphometric outcome after allograft transplantation of the sciatic nerve in rats at low doses of FK 506. Functional recovery and quantitative assessment of myelination were investigated in un-operated controls, in rats receiving isograft transplants without FK 506 treatment and in rats receiving allograft transplants with FK 506 treatment (0.1mg/kg and 0.2mg/kg per day). Walking-track analysis at 4, 8, 12 and 16 weeks post-operation revealed significant functional recovery in allograft with FK 506 (0.1mg/kg) compared with other groups, although levels of the un-operated controls were not reached. At 16 weeks, myelination of nerve sections from FK 506 (0.1mg/kg)-treated and un-operated animals did not differ significantly. There was significantly less effect of the 0.2mg/kg dose than of the 0.1mg/kg dose, both in the histomorphological outcome and in the functional outcome. These findings indicate that higher doses of FK 506 are not necessary for nerve regeneration, and low-dose administration could be acceptable for clinical settings in future.

Correlation of three sciatic functional indices with histomorphometric findings in a rat sciatic nerve allograft repair model.

Walking track analysis was used to measure global functional recovery following sciatic nerve injury. The correlation of morphologic outcome and different sciatic functional indices (SFIs) depends on different variables. The objective of this study was to compare three different SFIs and their correlation with histomorphometric findings in a sciatic nerve allograft repair model in the rat without (group I, n = 8) or with (group II, n = 8) daily intramuscular administration of 0.1 mg/kg FK 506. The correlation of SFIs with each other and with the myelin basic protein (MBP) density of nerve sections proximal, median, and distal to sciatic nerve grafts (1.5 cm) at 4, 8, 12, and 16 weeks postoperation (p.o.) was calculated, and unoperated animals served as controls (n = 8). Significant differences between SFIs calculated for experimental groups I and II at 12 and 16 weeks p.o. suggested that superior functional nerve recovery occurred in group II. However, there were significant differences between all SFIs at 16 weeks p.o. in group II, whereas only differences between SFI 1 and SFI 2 + 3 occurred in group I. SFIs of group II did not reach the values of the unoperated group. There were significant differences between the histomorphometric outcomes of groups I and II. There was no significant difference of MBP density between group II and the unoperated group, suggesting complete morphologic recovery. In conclusion, we found significant correlation between the MBP densities of groups I and II and all SFIs, suggesting a close relationship between histomorphometric and functional findings. (c) 2009 Wiley-Liss, Inc. Microsurgery, 2009.

Histomorphological and functional impacts of postoperative motor training in rats after allograft sciatic nerve transplantation under low-dose FK 506.

This study aimed to determine the effect of motor training on recovery after nerve transplantation under low-dose FK 506. Rats (n=30) of two strains were randomly assigned to three groups. Group I served as untreated controls; groups II and III received allograft transplants for reconstruction of the sciatic nerve and FK 506 (0.1 mg/kg/d). Nonoperated limbs served as intra-animal controls. Group III received postoperative motor training. Functional and histomorphological outcomes were assessed by walking track analysis and by blob analysis for myelinization of nerve sections. Regeneration occurred in both groups II and III. The control sections of the nonoperated limbs in group III showed significantly higher myelinization compared with group I and II; regeneration of the operated side was superior in group II. With regard to postoperative motor training, no benefit could be seen; however, the impact of postoperative motor training on the nonoperated limb were identified.

Thalamo-telencephalic pathways in the fire-bellied toad Bombina orientalis.

It was suggested that among extant vertebrates, anuran amphibians display a brain organization closest to the ancestral tetrapod condition, and recent research suggests that anuran brains share important similarities with the brains of amniotes. The thalamus is the major source of sensory input to the telencephalon in both amphibians and amniote vertebrates, and this sensory input is critical for higher brain functions. The present study investigated the thalamo-telencephalic pathways in the fire-bellied toad Bombina orientalis, a basal anuran, by using a combination of retrograde tract tracing and intracellular injections with the tracer biocytin. Intracellular labeling revealed that the majority of neurons in the anterior and central thalamic nuclei project to multiple brain targets involved in behavioral modulation either through axon collaterals or en passant varicosities. Single anterior thalamic neurons target multiple regions in the forebrain and midbrain. Of note, these neurons display abundant projections to the medial amygdala and a variety of pallial areas, predominantly the anterior medial pallium. In Bombina, telencephalic projections of central thalamic neurons are restricted to the dorsal striato-pallidum. The bed nucleus of the pallial commissure/thalamic eminence similarly targets multiple brain regions including the ventral medial pallium, but this is accomplished through a higher variety of distinct neuron types. We propose that the amphibian diencephalon exerts widespread influence in brain regions involved in behavioral modulation and that a single dorsal thalamic neuron is in a position to integrate different sensory channels and distribute the resulting information to multiple brain regions.

Extremely high-power tongue projection in plethodontid salamanders.

Many plethodontid salamanders project their tongues ballistically at high speed and for relatively great distances. Capturing evasive prey relies on the tongue reaching the target in minimum time, therefore it is expected that power production, or the rate of energy release, is maximized during tongue launch. We examined the dynamics of tongue projection in three genera of plethodontids (Bolitoglossa, Hydromantes and Eurycea), representing three independent evolutionary transitions to ballistic tongue projection, by using a combination of high speed imaging, kinematic and inverse dynamics analyses and electromyographic recordings from the tongue projector muscle. All three taxa require high-power output of the paired tongue projector muscles to produce the observed kinematics. Required power output peaks in Bolitoglossa at values that exceed the greatest maximum instantaneous power output of vertebrate muscle that has been reported by more than an order of magnitude. The high-power requirements are likely produced through the elastic storage and recovery of muscular kinetic energy. Tongue projector muscle activity precedes the departure of the tongue from the mouth by an average of 117 ms in Bolitoglossa, sufficient time to load the collagenous aponeuroses within the projector muscle with potential energy that is subsequently released at a faster rate during tongue launch.

Distribution of GABA, glycine, and glutamate in neurons of the medulla oblongata and their projections to the midbrain tectum in plethodontid salamanders.

In the medulla oblongata of plethodontid salamanders, GABA-, glycine-, and glutamate-like immunoreactivity (ir) of neurons was studied. Combined tracing and immunohistochemical experiments were performed to analyze the transmitter content of medullary nuclei with reciprocal connections with the tectum mesencephali. The distribution of transmitters differed significantly between rostral and caudal medulla; dual or triple localization of transmitters was present in somata throughout the rostrocaudal extent of the medulla. Regarding the rostral medulla, the largest number of GABA- and gly-ir neurons was found in the medial zone. Neurons of the nucleus reticularis medius (NRM) retrogradely labeled by tracer application into the tectum revealed predominantly gly-ir, often colocalized with glu-ir. The NRM appears to be homologous to the mammalian gigantocellular reticular nucleus, and its glycinergic projection is most likely part of a negative feedback loop between medulla and tectum. Neurons of the dorsal and vestibular nucleus projecting to the tectum were glu-ir and often revealed additional GABA- and/or gly-ir in the vestibular nucleus. Regarding the caudal medulla, the highest density of GABA- and gly-ir cells was found in the lateral zone. Differences in the neurochemistry of the rostral versus caudal medulla appear to result from the transmitter content of projection nuclei in the rostral medulla and support the idea that the rostral medulla is involved in tecto-reticular interaction. Our results likewise underline the role of the NRM in visual object selection and orientation as suggested by behavioral studies and recordings from tectal neurons.

Evolution of the brain and intelligence.

Intelligence has evolved many times independently among vertebrates. Primates, elephants and cetaceans are assumed to be more intelligent than 'lower' mammals, the great apes and humans more than monkeys, and humans more than the great apes. Brain properties assumed to be relevant for intelligence are the (absolute or relative) size of the brain, cortex, prefrontal cortex and degree of encephalization. However, factors that correlate better with intelligence are the number of cortical neurons and conduction velocity, as the basis for information-processing capacity. Humans have more cortical neurons than other mammals, although only marginally more than whales and elephants. The outstanding intelligence of humans appears to result from a combination and enhancement of properties found in non-human primates, such as theory of mind, imitation and language, rather than from 'unique' properties.

Activation patterns of the tongue-projector muscle during feeding in the imperial cave salamander Hydromantes imperialis.

Salamanders of the genus Hydromantes project their tongues the greatest distance of any amphibian to capture prey, up to 80% of body length or approximately 6 cm in an adult individual. During tongue projection on distant prey, the tongue is shot ballistically and the tongue skeleton leaves the body of the salamander entirely. We investigated an aspect of the motor control of this remarkable behavior by examining electromyographic patterns within different regions of the tongue-projector muscle, the subarcualis rectus (SAR). SAR activation is strongly modulated, and features of this modulation can be predicted by tongue-projection distance (i.e. prey distance). The strap-like buccal portion of the SAR is always activated first and for the longest duration, compared to any other region. It is in a position to transmit force generated by the posterior SAR to the floor of the mouth, where it originates. The posterior SAR encompasses and applies force to the epibranchial of the tongue skeleton, and its activation pattern gradually changes from a posterior-to-anterior wave of activation onset during short-distance projection to an all-at-once pattern during the most extreme long-distance (ballistic) projection. The duration of activity and EMG area of each recorded region of the SAR increase with increasing prey distance, showing greater muscle recruitment during long-distance projection. No effect of prey-capture success was observed in the EMG patterns, indicating that SAR activation is controlled in a feed-forward manner.

Dorsal striatopallidal system in anurans.

The dorsal striatopallidal system of tetrapods consists of the dorsal striatum (caudate-putamen in mammals) and the dorsal pallidum. Although the existence of striatal and pallidal structures has been well documented in anuran amphibians, the exact boundaries of these structures have so far been a matter of debate. To delineate precisely the dorsal striatopallidal system of anurans, we used quantitative analysis of leucine-enkephalin immunohistochemistry (in Bombina orientalis, Discoglossus pictus, Xenopus laevis, and Hyla versicolor), retrograde neurobiotin tracing studies (injections in the central and ventromedial thalamic nuclei in H. versicolor), and double-labeling tracing studies (injections in the lateral forebrain bundle and the caudal striatum in B. orientalis). Immunohistochemistry revealed that enkephalin-positive neurons are located mainly in the rostral and intermediate striatum. Neurobiotin tracing studies demonstrated that neurons projecting to the central and ventromedial thalamic nuclei are found in the intermediate and caudal striatum. Double-labeling studies revealed that the population of neurons in the rostral and intermediate striatum innervating the caudal striatum is separated from neurons projecting into the lateral forebrain bundle. Neurons that project to both the caudal striatum and the lateral forebrain bundle are found only in the dorsal part of the intermediate striatum. Taken together, our results suggest that the rostral striatum of anurans is homologous to the striatum proper of mammals, whereas the caudal striatum is comparable to the dorsal pallidum. The intermediate striatum represents a transition area between the two structures.

Morphology, axonal projection pattern, and responses to optic nerve stimulation of thalamic neurons in the fire-bellied toad Bombina orientalis.

Intracellular recording and biocytin labeling were carried out in the fire-bellied toad Bombina orientalis to study the morphology and axonal projections of thalamic (TH) neurons and their responses to electrical optic nerve stimulation. Labeled neurons (n = 142) were divided into the following groups: TH1 neurons projecting to the dorsal striatum; TH2 neurons projecting to the amygdala, nucleus accumbens, and septal nuclei; TH3 neurons projecting to the medial or dorsal pallium; TH4 neurons with projections ascending to the dorsal striatum or ventral striatum/amygdala and descending to the optic tectum, tegmentum, and rostral medulla oblongata; TH5 neurons with projections to the tegmentum, rostral medulla oblongata, prectectum, or tectum; and TH6 neurons projecting to the hypothalamus. TH1 neurons are found in the central, TH2 neurons in the anterior and central, TH3 neurons in the anterior dorsal nucleus, and TH4 and TH5 neurons in the posterior dorsal or ventral nucleus. Neurons with descending projections arborize in restricted parts of retinal afferents; neurons with ascending projections do not substantially arborize within retinal afferents. At electrical optic nerve stimulation, neurons in the ventral thalamus respond with excitation at latencies of 10.8 msec; one-third of them follow repetitive stimulation and possibly are monosynaptically driven. Neurons in the dorsal thalamus respond mostly with inhibition at latencies of 42.3 msec and are polysynaptically driven. This corroborates the view that neurons in the dorsal thalamus projecting to the telencephalon receive no substantial direct retinal input and that the thalamopallial pathway of amphibians is not homologous to the mammalian retinogeniculocortical pathway.