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Neurons can change their classical neurotransmitters during ontogeny, sometimes going through stages of dual release. Here, we explored the development of the neurotransmitter identity of neurons of the avian nucleus isthmi parvocellularis (Ipc), whose axon terminals are retinotopically arranged in the optic tectum (TeO) and exert a focal gating effect upon the ascending transmission of retinal inputs. Although cholinergic and glutamatergic markers are both found in Ipc neurons and terminals of adult pigeons and chicks, the mRNA expression of the vesicular acetylcholine transporter, VAChT, is weak or absent. To explore how the Ipc neurotransmitter identity is established during ontogeny, we analyzed the expression of mRNAs coding for cholinergic (ChAT, VAChT, and CHT) and glutamatergic (VGluT2 and VGluT3) markers in chick embryos at different developmental stages. We found that between E12 and E18, Ipc neurons expressed all cholinergic mRNAs and also VGluT2 mRNA; however, from E16 through posthatch stages, VAChT mRNA expression was specifically diminished. Our ex vivo deposits of tracer crystals and intracellular filling experiments revealed that Ipc axons exhibit a mature paintbrush morphology late in development, experiencing marked morphological transformations during the period of presumptive dual vesicular transmitter release. Additionally, although ChAT protein immunoassays increasingly label the growing Ipc axon, this labeling was consistently restricted to sparse portions of the terminal branches. Combined, these results suggest that the synthesis of glutamate and acetylcholine, and their vesicular release, is complexly linked to the developmental processes of branching, growing and remodeling of these unique axons.

AP-2 is a family of transcription factors involved in many aspects of development, cell differentiation, and regulation of cell growth and death. AP-2δ is a member of this group and specific gene expression patterns are required in the adult mouse brain for the development of parts of the inferior colliculus (IC), as well as the cortex, dorsal thalamus, and superior colliculus. The midbrain is one of the central areas in the brain where multimodal integration, i.e., integration of information from different senses, occurs. Previous data showed that AP-2δ-deficient mice are viable but due to increased apoptosis at the end of embryogenesis, lack part of the posterior midbrain. Despite the absence of the IC in AP-2δ-deficient mice, these animals retain at least some higher auditory functions. Neuronal responses to tones in the neocortex suggest an alternative auditory pathway that bypasses the IC. While sufficient data are available in mammals, little is known about AP-2δ in chickens, an avian model for the localization of sounds and the development of auditory circuits in the brain. Here, we identified and localized AP-2δ expression in the chicken midbrain during embryogenesis. Our data confirmed the presence of AP-2δ in the inferior colliculus and optic tectum (TeO), specifically in shepherd’s crook neurons, which are an essential component of the midbrain isthmic network and involved in multimodal integration. AP-2δ expression in the chicken midbrain may be related to the integration of both auditory and visual afferents in these neurons. In the future, these insights may allow for a more detailed study of circuitry and computational rules of auditory and multimodal networks.

The midbrain is a subcortical area involved in central functions such as integrating sensory modalities, movement initiation and bottom-up and top-down attention. In chicken, the midbrain roof is termed optic tectum (TeO) and consists of 15 layers with distinct in- and output regions. Visual input targets the superficial layers, while auditory input terminates in deeper layers. It has been shown that ablation of sensory epithelia leads to changes in the cellular patterning and structural organization of the sensory pathways. For the tectum, ablation of the eye anlagen was shown to affect retinorecipient neurons. While the gross morphology remained intact after enucleation, the shape of dendritic endings was changed presumably due to missing presynaptic input during synaptic pruning. We investigated the effect of deafferentation in a multisensory cell type, the Shepherd's crook neuron (SCN) in the TeO. SCNs have distinct dendritic branches in retinorecipient layers (superficial layers 1 to 5 and 7) and in layers where auditory input terminates. To assess whether removal of a single sensory input only affects the dendrites recipient for that input, we removed the eye anlagen and retrogradely labeled SCNs later in embryogenesis to visualize the morphology in lesioned and non-lesioned embryos. We found no changes in the gross morphology or in the basal dendrites, but an altered growth of the fine structures at the apical dendrite of SCNs in the retinorecipient layers. Our data indicate that the neuronal morphology of SCNs is mostly predefined before retinal innervation affect the fine structure.

The midbrain is an important subcortical area involved in distinct functions such as multimodal integration, movement initiation, bottom-up, and top-down attention. Our group is particularly interested in cellular computation of multisensory integration. We focus on the visual part of the avian midbrain, the optic tectum (TeO, counterpart to mammalian superior colliculus). This area has a layered structure with the great advantage of distinct input and output regions. In chicken, the TeO is organized in 15 layers where visual input targets the superficial layers while auditory input terminates in deeper layers. One specific cell type, the Shepherd's crook neuron (SCN), extends dendrites in both input regions. The characteristic feature of these neurons is the axon origin at the apical dendrite. The molecular identity of this characteristic region and thus, the site of action potential generation are of particular importance to understand signal flow and cellular computation in this neuron. We present immunohistochemical data of structural proteins (NF200, Ankyrin G, and Myelin) and ion channels (Pan-Nav , Nav 1.6, and Kv 3.1b). NF200 is strongly expressed in the axon. Ankyrin G is mainly expressed at the axon initial segment (AIS). Myelination starts after the AIS as well as the distribution of Nav channels on the axon. The subtype Nav 1.6 has a high density in this region. Kv 3.1b is restricted to the soma, the primary neurite and the axon branch. The distribution of functional molecules in SCNs provides insight into the information flow and the integration of sensory modalities in the TeO of the avian midbrain.

The avian centrifugal visual system, which projects from the brain to the retina, has been intensively studied in several Neognathous birds that have a distinct isthmo-optic nucleus (ION). However, birds of the order Palaeognathae seem to lack a proper ION in histologically stained brain sections. We had previously reported in the palaeognathous Chilean Tinamou (Nothoprocta perdicaria) that intraocular injections of Cholera Toxin B subunit retrogradely label a considerable number of neurons, which form a diffuse isthmo-optic complex (IOC). In order to better understand how this IOC-based centrifugal visual system is organized, we have studied its major components by means of in vivo and in vitro tracing experiments. Our results show that the IOC, though structurally less organized than an ION, possesses a dense core region consisting of multipolar neurons. It receives afferents from neurons in L10a of the optic tectum, which are distributed with a wider interneuronal spacing than in Neognathae. The tecto-IOC terminals are delicate and divergent, unlike the prominent convergent tecto-ION terminals in Neognathae. The centrifugal IOC terminals in the retina are exclusively divergent, resembling the terminals from “ectopic” centrifugal neurons in Neognathae. We conclude that the Tinamou's IOC participates in a comparable general IOC-retina-TeO-IOC circuitry as the neognathous ION. However, the connections between the components are structurally different and their divergent character suggests a lower spatial resolution. Our findings call for further comparative studies in a broad range of species for advancing our understanding of the evolution, plasticity and functional roles of the avian centrifugal visual system.

Sharp wave ripples (SWR) represent one of the most synchronous population patterns in the mammalian brain. Although SWRs are highly conserved throughout mammalian evolution, the existence of SWRs in non-mammalian species remains controversial. We reexamined the existence of avian SWRs by recording the brain activity during sleep and under anesthesia in two species of birds, the zebra finch and the chicken. Electrophysiological recordings using silicon probes implanted in the avian telencephalon revealed highly dynamic switching between high and low delta phases during sleep. High delta phases were composed of large-amplitude, negative deflections (sharp waves) that coincided with a high frequency oscillation (ripple). Correlation analysis revealed that these events were highly synchronous and spanned a large anatomical range of the avian telencephalon. Finally, detailed spike analysis revealed that an increase in the population spiking activity coincided with the occurrence of SWRs, that this spiking activity occurred in specific sequences of spike patterns locked to the SWRs, and that the mean population spiking activity peaked prior to the trough of the negative deflection. These results provide the first evidence of avian SWRs during natural sleep and under anesthesia, and suggest that the evolutionary origin of SWR activity may precede the mammalian-sauropsid bifurcation.

Echolocating bats evolved a sophisticated biosonar imaging system that allows for a life in dim-light habitats. However, especially for far-range operations such as homing, bats can support biosonar by vision. Large eyes and a retina that mainly consists of rods are assumed to be the optical adjustments that enable bats to use visual information at low light levels. In addition to optical mechanisms, many nocturnal animals evolved neural adaptations such as elongated integration times or enlarged spatial sampling areas to further increase the sensitivity of their visual system by temporal or spatial summation of visual information. The neural mechanisms that underlie the visual capabilities of echolocating bats have, however, so far not been investigated. To shed light on spatial and temporal response characteristics of visual neurons in an echolocating bat, Phyllostomus discolor, we recorded extracellular multiunit activity in the retino-recipient superficial layers of the superior colliculus (SC). We discovered that response latencies of these neurons were generally in the mammalian range, whereas neural spatial sampling areas were unusually large compared to those measured in the SC of other mammals. From this we suggest that echolocating bats likely use spatial but not temporal summation of visual input to improve visual performance under dim-light conditions. Furthermore, we hypothesize that bats compensate for the loss of visual spatial precision, which is a byproduct of spatial summation, by integration of spatial information provided by both the visual and the biosonar systems. Given that knowledge about neural adaptations to dim-light vision is mainly based on studies done in non-mammalian species, our novel data provide a valuable contribution to the field and demonstrate the suitability of echolocating bats as a nocturnal animal model to study the neurophysiological aspects of dim-light vision.

Lugaro cells are inhibitory interneurons found in the upper granular layer of the cerebellar cortex, just below or within the Purkinje cell layer. They are characterized by (1) a fusiform soma oriented in the parasagittal plane, (2) two pairs of dendrites emanating from opposite ends of the soma, (3) innervation from Purkinje cell collaterals, and (4) an axon that projects into the molecular layer akin to granular cell parallel fibers. Lugaro cells have been described in mammals, but not in other vertebrate classes, save one report in teleost fish. Here, we propose the existence of Lugaro cells in the avian cerebellum based on the morphological characteristics and connectivity described above. Immunohistochemical staining against the calcium binding protein secretagogin (SCGN) revealed Lugaro-like cells in the pigeon cerebellum. Some SCGN-labeled cells exhibit fusiform somata and dendrites parallel to the Purkinje cell layer in the parasagittal plane, as well as long axons that project into the molecular layer and travel alongside parallel fibers in the coronal plane. While mammalian Lugaro cells are known to express calretinin, the SCGN-labeled cells in the pigeon do not. SCGN-labeled cells also express glutamic acid decarboxylase, confirming their inhibitory function. Calbindin labeling revealed Purkinje cell terminals surrounding the SCGN-expressing cells. Our results suggest that Lugaro cells are more widespread among vertebrates than previously thought and may be a characteristic of the cerebellum of all vertebrates.

While in birds and mammals the cerebellum is a highly convoluted structure that consists of numerous transverse lobules, in most amphibians and reptiles it consists of only a single unfolded sheet. Orthogonal to the lobules, the cerebellum is comprised of sagittal zones that are revealed in the pattern of afferent inputs, the projection patterns of Purkinje cells, and Purkinje cell response properties, among other features. The expression of several molecular markers, such as aldolase C, is also parasagittally organized. Aldolase C, also known as zebrin II (ZII), is a glycolytic enzyme expressed in the cerebellar Purkinje cells of the vertebrate cerebellum. In birds, mammals, and some lizards (Ctenophoresspp.), ZII is expressed in a heterogenous fashion of alternating sagittal bands of high (ZII+) and low (ZII–) expression Purkinje cells. In contrast, turtles and snakes express ZII homogenously (ZII+) in their cerebella, but the pattern in crocodilians is unknown. Here, we examined the expression of ZII in two crocodilian species (Crocodylus niloticus and Alligator mississippiensis) to help determine the evolutionary origin of striped ZII expression in vertebrates. We expected crocodilians to express ZII in a striped (ZII+/ZII–) manner because of their close phylogenetic relationship to birds and their larger and more folded cerebellum compared to that of snakes and turtles. Contrary to our prediction, all Purkinje cells in the crocodilian cerebellum had a generally homogenous expression of ZII (ZII+) rather than clear ZII+/– stripes. Our results suggest that either ZII stripes were lost in three groups (snakes, turtles, and crocodilians) or ZII stripes evolved independently three times (lizards, birds, and mammals).

Cocaine- and amphetamine-regulated transcript peptides (CARTp) are neuropeptides that act as neurotransmitters in the brain of vertebrates. The expression of CARTp has been characterized in teleosts, amphibians, and several mammalian species, but comparative data in reptiles and birds are nonexistent. In this study, we show the distribution of immunoreactivity against CART peptides (CARTp-ir) in the brains of two bird species: the pigeon (Columba livia) and zebra finch (Taeniopygia guttata). We found CARTp-ir cells and terminals in the brains of both, but no major differences between the two species. As in mammals, teleost fish, and amphibians, CARTp-ir terminals and cells were abundant in subpallial regions, particularly the striatum and nucleus accumbens. We also found CARTp-ir cells and terminals in the hypothalamus, and a large number of CARTp-ir terminals in the substantia nigra, ventral tegmental area, periaqueductal gray, parabrachial nucleus, and dorsal vagal complex. However, in contrast to other vertebrates, CARTp-ir was not found in the olfactory bulb. In addition there was almost no CARTp-ir in the pallium or the hippocampal formation, and little CARTp-ir in the cerebellum. The conserved expression of CARTp in the subpallium, hypothalamus, and dorsal vagal complex of birds suggests that some of the functions of CARTp, such as regulation of food intake and interactions with the social control network and mesolimbic reward system, are conserved among vertebrates. J. Comp. Neurol. 524:3747-3773, 2016. © 2016 Wiley Periodicals, Inc.

Background: The epithelial layer of the nasal mucosa is the first barrier for drug permeation during intranasal drug delivery. With increasing interest for intranasal pathways, adequate in vitro models are required. Here, porcine olfactory (OEPC) and respiratory (REPC) primary cells were characterised against the nasal tumour cell line RPMI 2650. Methods: Culture conditions for primary cells from porcine nasal mucosa were optimized and the cells characterised via light microscope, RT-PCR and immunofluorescence. Epithelial barrier function was analysed via transepithelial electrical resistance (TEER), and FITC-dextran was used as model substance for transepithelial permeation. Beating cilia necessary for mucociliary clearance were studied by immunoreactivity against acetylated tubulin. Results: OEPC and REPC barrier models differ in TEER, transepithelial permeation and MUC5AC levels. In contrast, RPMI 2650 displayed lower levels of MUC5AC, cilia markers and TEER, and higher FITC-dextran flux rates. Conclusion: To screen pharmaceutical formulations for intranasal delivery in vitro, translational mucosal models are needed. Here, a novel and comprehensive characterisation of OEPC and REPC against RPMI 2650 is presented. The established primary models display an appropriate model for nasal mucosa with secreted MUC5AC, beating cilia and a functional epithelial barrier, which is suitable for long-term evaluation of sustained release dosage forms.

The optic tectum (TeO), or superior colliculus, is a multisensory midbrain center that organizes spatially orienting responses to relevant stimuli. To define the stimulus with the highest priority at each moment, a network of reciprocal connections between the TeO and the isthmi promotes competition between concurrent tectal inputs. In the avian midbrain, the neurons mediating enhancement and suppression of tectal inputs are located in separate isthmic nuclei, facilitating the analysis of the neural processes that mediate competition. A specific subset of radial neurons in the intermediate tectal layers relay retinal inputs to the isthmi, but at present it is unclear whether separate neurons innervate individual nuclei or a single neural type sends a common input to several of them. In this study, we used in vitro neural tracing and cell-filling experiments in chickens to show that single neurons innervate, via axon collaterals, the three nuclei that comprise the isthmotectal network. This demonstrates that the input signals representing the strength of the incoming stimuli are simultaneously relayed to the mechanisms promoting both enhancement and suppression of the input signals. By performing in vivo recordings in anesthetized chicks, we also show that this common input generates synchrony between both antagonistic mechanisms, demonstrating that activity enhancement and suppression are closely coordinated. From a computational point of view, these results suggest that these tectal neurons constitute integrative nodes that combine inputs from different sources to drive in parallel several concurrent neural processes, each performing complementary functions within the network through different firing patterns and connectivity.

Pitvipers have a specialized sensory system in the upper jaw to detect infrared (IR) radiation. The bilateral pit organs resemble simple pinhole cameras that map IR objects onto the sensory epithelium as blurred representations of the environment. Trigeminal afferents transmit information about changing temperature patterns as neuronal spike discharge in a topographic manner to the hindbrain nucleus of the lateral descending trigeminal tract (LTTD). A presumed, yet so far unknown neuronal connectivity within this central nucleus exerts a synaptic computation that constrains the relatively large receptive field of primary afferent fibers. Here, we used intracellular recordings of LTTD neurons in isolated rattlesnake brains to decipher the spatio-temporal pattern of excitatory and inhibitory responses following electrical stimulation of single and multiple peripheral pit organ-innervating nerve branches. The responses of individual neurons consisted of complex spike sequences that derived from spatially and temporally specific interactions between excitatory and inhibitory synaptic inputs from the same as well as from adjacent peripheral nerve terminal areas. This pattern complies with a central excitation that is flanked by a delayed lateral inhibition, thereby enhancing the contrast of IR sensory input, functionally reminiscent of the computations for contrast enhancement in the peripheral visual system.

The ability to detect motion is crucial for the survival of animals. In the avian optic tectum, motion-sensitive output neurons in the stratum griseum centrale have large dendritic fields and receive direct retinal input at their distal dendrites (bottlebrush endings). It has been hypothesized that the activation of each ending elicits a burst in the neuron. Thus, an object moving across the receptive field would lead to a fixed number of bursts, independent of movement speed. However, experimental confirmation of this hypothesis is still missing. We measured the response of tectal neurons to moving stimuli in vivo and found that in 'fast-bursting' units (~500 Hz within burst), the number of bursts was independent of stimulus speed. These results indicate that the number of bursts might indeed be related to the sequential activation of the bottlebrush endings by visual stimuli.

Most systematic studies of the avian visual system have focused on Neognathous species, leaving virtually unexplored the Palaeognathae, comprised of the flightless ratites and the South American tinamous. We investigated the visual field, the retinal topography, and the pattern of retinal and centrifugal projections in the Chilean tinamou, a small Palaeognath of the family Tinamidae. The tinamou has a panoramic visual field with a small frontal binocular overlap of 20°. The retina possesses three distinct topographic specializations: a horizontal visual streak, a dorsotemporal area, and an area centralis with a shallow fovea. The maximum ganglion cell density is 61,900/ mm(2) , comparable to Falconiformes. This would provide a maximal visual acuity of 14.0 cycles/degree, in spite of relatively small eyes. The central retinal projections generally conform to the characteristic arrangement observed in Neognathae, with well-differentiated contralateral targets and very few ipsilateral fibers. The centrifugal visual system is composed of a considerable number of multipolar centrifugal neurons, resembling the "ectopic" neurons described in Neognathae. They form a diffuse nuclear structure, which may correspond to the ancestral condition shared with other sauropsids. A notable feature is the presence of terminals in deep tectal layers 11-13. These fibers may represent either a novel retinotectal pathway or collateral branches from centrifugal neurons projecting to the retina. Both types of connections have been described in chicken embryos. Our results widen the basis for comparative studies of the vertebrate visual system, stressing the conserved character of the visual projections' pattern within the avian clade.

Pit vipers detect infrared (IR) radiation with loreal pit organs [1] that are connected to the hindbrain by trigeminal nerve fibers [2-4]. The pattern of central afferent termination forms a topographical representation of the sensory periphery within the nucleus of the lateral descending trigeminal tract (LTTD) [4-7]. All LTTD neurons project to another specialized, ipsilateral hindbrain area, the nucleus reticularis caloris (RC) [8-11], before IR signals are integrated with visual signals in the optic tectum [12, 13]. Pit-organ-innervating afferent fibers provoke in individual LTTD neurons a direct, robust spike activity upon peripheral activation [7, 14]. This discharge is truncated by an indirect, delayed synaptic inhibition from afferent fibers of adjacent sensory areas through parallel microcircuitry that converges with afferent fibers onto the same target neurons [7]. Here, we determined the impact of this interaction on IR contrast enhancement and/or motion detection in LTTD and RC neurons using isolated whole-brain preparations of rattlesnakes with intact pit organs. Simulated and real IR source motion provoked weak directional tuning of the discharge in LTTD neurons and RC neurons expressed a strong, motion-direction-differentiating activity. The hierarchically increasing motion sensitivity potentially derives from a direction-specific inhibition or spike frequency adaptation of LTTD neuronal discharge that becomes further pronounced by convergent projections onto individual RC neurons. The emerging signaling pattern complies with contrast enhancement (LTTD) and extraction of movement-related signals (RC), thereby forming a motion detection mechanism that encodes moving IR sources relative to the ambient temperature [14].

The avian inferior colliculus (IC), also referred to as the nucleus mesencephalicus lateralis pars dorsalis (MLd), is an auditory midbrain nucleus that converges auditory cues from tonotopically organized brainstem nuclei. This information is relayed onto the optic tectum on the one hand and to nucleus ovoidalis on the other hand. Morphologically, there has been considerable debate about the number and nomenclature of the subnuclei within the IC. Here, we provide morphological characteristics of single cells in five IC subnuclei in chicken. The cellular structure within the IC was studied by whole-cell patch technique and biocytin iontophoresis. In addition, histological staining was performed, to delineate the borders between subnuclei of the IC. We were able to discriminate between 5 subnuclei: the core of the central nucleus (ICCc), the medial and lateral shell of the central nucleus (ICCms and ICCls), the external nucleus (ICX) and the superficial nucleus (ICS) of the IC. Our findings suggest the existence of at least two different morphologies of neurons with two subtypes each. The IC in chicken is a largely homogenous nucleus in terms of neuronal anatomy on a cellular level. However, its compartmentation into diversified subnuclei with different neurophysiological characteristics suggests a complex system to process auditory information. The auditory system in chicken is not as hypertrophied as in specialists such as the barn owl, but appears to have comparable connectivity and cellular morphology.

We present a method to identify specific sub-populations of auditory neurons in a mixed primary cell culture of the chicken brainstem, allowing the study of individual neurons with a known identity in vitro. To label specific afferent cell types, we injected retrograde tracers (dextrans coupled to fluorescent dyes) into either the mid-line or the superior olivary nuclei (SON) of the isolated chicken brainstem in vitro. Mid-line injections resulted in stable labelling of neurons of the nucleus magnocellularis (NM), whereas injections into the SON retrogradely labelled neurons of the nucleus laminaris (NL). The fluorescent label survives the dissociation procedure and is detectable for at least 1 week in vitro. Only about 0.1% of all cells in vitro are pre-labelled. The auditory identity of the pre-labelled neurons was confirmed with calretinin immunocytochemistry and electrophysiological recordings, where the cells had typical firing patterns of auditory brainstem neurons. In the future, this method can be combined with single cell PCR to match nuclear origin, firing patterns and the expression of functional molecules in vitro.

Neurons in the auditory hindbrain pathway of the chicken are physiologically and morphologically highly specialized. It remains unclear to what extent independent differentiation vs. activity-dependent mechanisms determines the development of this system. To address this question we established a primary culture system of the early auditory hindbrain neurons. Primary cultures of neurons from nucleus magnocellularis and nucleus laminaris were prepared from embryonic day 6.5 chicken. These cells developed in culture under serum-free conditions for up to 15 days. Immunocytochemical staining and whole-cell patch recordings were used to characterize the development of the neurons. A stable expression of the calcium-binding protein calretinin, which serves as a characteristic marker of the auditory pathway, was found at all stages. A voltage-gated potassium channel (Kv3.1b) with a specific function in the auditory system was also expressed after about 1 week in culture. Electrophysiological recordings showed a general maturation of the neuronal phenotype as reflected by an increase in the mean resting membrane potential, a decrease in the mean input resistance as well as a maturation of action potential parameters. Four groups of neurons that generate action potentials could be distinguished. One of these showed the phasic firing pattern of auditory brainstem neurons known from slice preparations. In older cultures we demonstrated functional synaptogenesis in vitro by recording postsynaptic activity elicited by extracellular stimulation and styryl dye loading of vesicles. Thus, isolated neurons from the auditory region of the avian brainstem differentiate to specific neuronal subtypes and autonomously develop synaptic connections in vitro.

The avian retino-tecto-rotundal pathway plays a central role in motion analysis and features complex connectivity. Yet, the relation between the pathway's structural arrangement and motion computation has remained elusive. For an important type of tectal wide-field neuron, the stratum griseum centrale type I (SGC-I) neuron, we quantified its structure and found a spatially sparse but extensive sampling of the retinal projection. A computational investigation revealed that these structural properties enhance the neuron's sensitivity to change, a behaviorally important stimulus attribute, while preserving information about the stimulus location in the SGC-I population activity. Furthermore, the SGC-I neurons project with an interdigitating topography to the nucleus rotundus, where the direction of motion is computed. We showed that, for accurate direction-of-motion estimation, the interdigitating projection of tectal wide-field neurons requires a two-stage rotundal algorithm, where the second rotundal stage estimates the direction of motion from the change in the relative stimulus position represented in the first stage.

Humans and other animals generally perceive motion independently of the cues that define the moving object. To understand the underlying mechanisms of this generalization of stimulus attributes, we have examined the cellular properties of avian wide-field tectal neurons that are sensitive to a variety of moving stimuli but not to static stationary stimuli. This in vitro study showed phasic signal transfer at the retinotectal synapse and binary dendritic responses to synaptic inputs that interact in a mutually exclusive manner in the postsynaptic tectal neuron. A model of the tectal circuitry predicts that these two cellular properties mediate sensitivity to a wide range of dynamic spatiotemporal stimuli, including moving stimuli, but not to static stationary stimuli in a tectal neuron. The computation that is independent of stimulus detail is initiated by tectal neurons and is completed by rotundal neurons that integrate outputs from multiple tectal neurons in a directionally selective manner.

The stable and specific locking-in of pre- and postsynaptic membranes in synaptogenesis may be mediated by integral membrane proteins, such as members of the cadherin family. Cadherins are ideal candidate molecules for mediating synaptic specificity because they are differentially expressed in functionally connected brain structures. We studied the expression of four classic cadherins (R-cadherin, N-cadherin, cadherin-6B and cadherin-7) at the synaptic level on the somata and the proximal neurites of identified neuron populations that were traced selectively in the developing chicken visual system. Three major findings were observed. (1) Synapses on somata of shepherd's crook cells of the optic tectum are associated preferentially with one cadherin subtype. (2) In an isthmic nucleus that contains a mixed population of cells expressing different cadherins, somatic synapses tend to express the same cadherin subtype as the rest of the cell. (3) In the oculomotor complex, two cadherin subtypes are expressed only by synapses on the axon hillock. However, another neuron type that projects from the tectum to the isthmic nucleus does not show such selective synaptic cadherin staining. Our findings support the idea that a cadherin-based adhesive mechanism can mediate synaptic specificity.

During development of the nervous system, neurotrophic interactions are essential for the synchronization of substructures such as retina and visual midbrain. However, morphological studies of postsynaptic elements after ablation of afferents are sparse. We investigated the effect of uni- and bilateral eye anlagen removal on identified projection neurons in the chicken optic tectum. Without retinal input, neurons in the stratum griseum centrale express their specific cell adhesions molecules, retain large dendritic fields and form specialized dendritic endings; however, the latter are deformed and extend over a much larger area. Our results show that even monosynaptically innervated tectal neurons develop largely independently from trophic retinal inputs and only become dependent on these after synaptic contact with retinal afferents.

The anuran torus semicircularis consists of several subnuclei that are part of the ascending auditory pathway as well as audiomotor interface structures. Additionally, recent anatomical studies suggest that the midbrain tegmentum is an integral part of the audiomotor network. To describe the chemoarchitecture of these nuclei, taking into account the toral subdivisions, we investigated the distribution of serotonin, leucine-enkephalin, substance P, tyrosine-hydroxylase, dopamine D2-receptor, parvalbumin, aspartate, GABA, and estrogen-binding protein-immunoreactivity in the midbrain of Bombina orientalis, Discoglossus pictus and Xenopus laevis. In the torus semicircularis, the highest density of immunoreactive fibers and terminals for all transmitters was found in the laminar nucleus. Parvalbumin-like immunoreactivity was highest in the principal nucleus, and D2-receptor-like immunoreactivity was uniformly distributed throughout the torus. In the tegmentum, axons and/or dendrites were stained with all antibodies except estrogen-binding protein. Additionally, heavily stained enkephalin and substance P-immunopositive fiber plexus were found in the lateral and dorsal tegmentum. The immunostainings revealed no qualitative differences between the three species. Immunopositive cell bodies were labeled in several brain areas, the connectivity of which with torus and tegmentum is discussed on the background of functional questions. The putative neuromodulatory innervation of both the laminar nucleus of the torus semicircularis and the tegmentum may be the anatomical basis for the influence of the animal's endogenous state on the behavioral reaction to sensory stimuli. These data corroborate earlier anatomical and physiological findings that the neurons of these nuclei are key elements in the audio-motor interface.

The widefield vertical neurons of the lower stratum griseum superficiale (SGS3) and upper stratum opticum (SO) of the superior colliculus provide an extrageniculate route for visual information to reach the pulvinar. Previous physiological studies indicate that SGS3/SO neurons have large receptive fields and respond to small moving stimuli. We sought to better characterize the dendritic morphology of SGS3/SO neurons with intracellular filling in slice preparations of the ground squirrel superior colliculus. We found that dendrites of widefield vertical cells end in monostratified arrays of spiny terminal specializations called "bottlebrush" dendritic endings. Two major subtypes of neurons are described. Type I neurons have somata restricted to the SGS3 and bottlebrush endings in the most superficial sublayer of the SGS. Type II neurons are found at the base of the SGS and in the upper SO, and have bottlebrush endings arrayed within the middle sublayers of the SGS. Bottlebrush endings may sample and integrate laminated afferents to the superior colliculus, and cellular subtypes may underlie multiple information streams within the tectopulvinar pathway. A similar dendritic morphology and projection pattern can be found in cells of the avian optic tectum that project upon the nucleus rotundus, a thalamic nucleus homologous to the mammalian caudal/inferior pulvinar. Because motion processing is a dominant feature of the avian tectorotundal pathway, the current results suggest that both dendritic morphology and motion processing are conserved features of widefield vertical cells in the tectopulvinar pathway of vertebrates.

Acoustic signals are extensively used for guiding various behaviors in frogs such as vocalization and phonotaxis. While numerous studies have investigated the anatomy and physiology of the auditory system, our knowledge of intrinsic properties and connectivity of individual auditory neurons remains poor. Moreover, the neural basis of audiomotor integration still has to be elucidated. We determined basic response patterns, dendritic arborization and axonal projection patterns of auditory midbrain units with intracellular recording and staining techniques in an isolated brain preparation. The subnuclei of the torus semicircularis subserve different tasks. The principal nucleus, the main target of the ascending auditory input, has mostly intrinsic neurons, i.e., their dendrites and axons are restricted to the torus itself. In contrast, neurons of the magnocellular and the laminar nucleus project to various auditory and non-auditory processing centers. The projection targets include thalamus, tegmentum, periaqueductal gray, medulla oblongata, and – in the case of laminar neurons – the spinal cord. Additionally, tegmental cells receive direct auditory input and project to various targets, including the spinal cord. Our data imply that both auditory and premotor functions are implemented in individual toral and tegmental neurons. Their axons constitute parallel descending pathways to several effector systems and might be part of the neural substrate for differential audiomotor integration.

The chicken optic tectum (TeO) and its mammalian counterpart, the superior colliculus, are important sensory integration centers. Multimodal information is represented in a topographic map, which plays a role in spatial attention and orientation movements. The TeO is organised in 15 layers with clear input and output regions, and further interconnected with the isthmic nuclei (NI), which modulate the response in a winner-takes-all fashion. While many studies have analysed tectal cell types and their modulation from the isthmic system physiologically, little is known about local network activity and its modulation in the tectum. We have recently shown with voltage-sensitive dye imaging that electrical stimulation of the retinorecipient layers results in a stereotypic response, which is under inhibitory control [S. Weigel & H. Luksch (2012) J. Neurophysiol., 107, 640–648]. Here, we analysed the contribution of acetylcholine (ACh) and the NI to evoked tectal responses using a pharmacological approach in a midbrain slice preparation. Application of the nicotinic ACh receptor (AChR) antagonist curarine increased the tectal response in amplitude, duration and lateral extent. This effect was similar but less pronounced when γ-aminobutyric acidA receptors were blocked, indicating interaction of inhibitory and cholinergic neurons. The muscarinic AChR antagonist atropine did not change the response pattern. Removal of the NI, which are thought to be the major source of cholinergic input to the TeO, reduced the response only slightly and did not result in a disinhibition. Based on the data presented here and the neuroanatomical literature of the avian TeO, we propose a model of the underlying local circuitry.

In the present study an isolated preparation of the complete anuran central nervous system (CNS) is described which can be kept alive for several days and allows tracing, immunohistochemical and electrophysiological studies. A simple perfusion chamber is being used in which the isolated CNS preparation is superfused with oxygenated Ringer. The use of an isolated CNS has many advantages including: (1) virtually all areas are easily accessible at the same time without having the problem of blood vessels that hinder access; (2) large lesions and massive tracer applications are possible without survival problems of the animal, and tracers will not be translocated by blood circulation; (3) since pulsations caused by the pressure changes of blood circulation do not occur, intracellular recordings are comparatively easy and stable; and (4) this approach offers the possibility of working on the same brain for several days by storing the preparation in a refrigerator overnight at low temperatures, thus allowing extensive utilization of a single preparation and reduction in the number of experimental animals required. Some applications to the anuran auditory system illustrate that the isolated anuran CNS is well-suited for a variety of neuroanatomical and physiological techniques.

SYNOPSIS. Anurans respond to different acoustic signals in a distinct manner. Moreover, acoustic behavior strongly depends on the inner state of the animal and the social context. Neuroanatomical studies as well as extra- and intracellular recordings were carried out to examine the problem of audio-motor interfacing. Acoustic signals are processed in a partly hierarchical and partly distributed manner in the anuran central nervous system. Features are represented in a topographical manner. Auditory maps in the midbrain torus semicircularis are read by a subset of neurons of the laminar and magnocellular subnuclei, which interconnect the auditory pathway and premotor centres. These, in turn, feed into neural networks controlling vocalization, locomotion, or autonomic functions. Our data on the cytoarchitectonic organization and connectivity of the interfacing neurons give rise to a population coding hypothesis which may explain the differential evaluation of acoustic signals. Additionally, immunohistochemical findings reveal strong neuromodulatory and hormonal input, especially onto interfacing neurons, suggesting intense influence on the physiological properties of these cells.

Topographically organized neurons represent multiple stimuli within complex visual scenes and compete for subsequent processing in higher visual centers. The underlying neural mechanisms of this process have long been elusive. We investigate an experimentally constrained model of a midbrain structure: the optic tectum and the reciprocally connected nucleus isthmi. We show that a recurrent antitopographic inhibition mediates the competitive stimulus selection between distant sensory inputs in this visual pathway. This recurrent antitopographic inhibition is fundamentally different from surround inhibition in that it projects on all locations of its input layer, except to the locus from which it receives input. At a larger scale, the model shows how a focal top-down input from a forebrain region, the arcopallial gaze field, biases the competitive stimulus selection via the combined activation of a local excitation and the recurrent antitopographic inhibition. Our findings reveal circuit mechanisms of competitive stimulus selection and should motivate a search for anatomical implementations of these mechanisms in a range of vertebrate attentional systems.

One strategy in localizing a sound source in the azimuthal plane is the comparison of arrival times of sound stimuli at the two ears. The processing of interaural time differences (ITDs) in the auditory brainstem was suggested by the Jeffress model in 1948. In chicks, binaural neurons in the nucleus laminaris (NL) receive input from both ipsilateral and contralateral nucleus magnocellularis (NM) neurons, with the axons of the latter acting as delay lines. A given neuron in the NL responds maximally to coinciding input from both NM neurons. To achieve maximum resolution of sound localization in the NL, the conduction velocity along these delay lines must be precisely tuned. Here, we examined the development of this velocity between embryonic days (E)12 and E18. Our optical imaging approach visualizes the contralateral delay lines along almost the complete NL of the chicken embryo. Optical imaging with the voltage-sensitive dye RH 795 showed no significant differences in the velocity between E12 and E15, but a significant increase from E15 to E18, at both 21 degrees C and 35 degrees C. Surprisingly, at 21 degrees C the conduction velocity in the dorso-lateral part of the NL was significantly higher compared to the situation in the ventro-medial part. The observed development in contralateral conduction velocity may be due to a developmental increase in myelination of the NM axons. Indeed, antibody staining against myelin-associated glycoprotein (alpha-MAG) showed no myelination of the NM axon branches within the NL at E12 and E15. On the other hand, a clear alpha-MAG immunoreactivity occurred at E18. Our results therefore describe the developmental physiological properties of the delay line in the chicken embryo.

In the auditory system voltage-activated currents mediated by potassium channels Kv1.1 and Kv3.1b and their interaction with sodium inward currents play a crucial role for computational function. However, it is unresolved how these potassium channels are developmentally regulated. We have therefore combined a biochemical investigation of Kv1.1 and Kv3.1b protein expression with electrophysiological recordings of membrane currents to characterize neuronal differentiation in the auditory brain stem of the chick. Differentiation in vitro was compared with cells prepared from corresponding embryonic stages in vivo. Using a computer model based on the empirical data we were then able to predict physiological properties of developing auditory brain stem neurons. In vivo Kv3.1b expression increased strongly between E10 and E14, a time of functional synaptogenesis in the auditory brainstem. We also found this increase of expression in vitro, again coinciding with synaptogenesis in the cultures. Whole-cell patch recordings revealed a corresponding increase of the (Kv3.1-like) high threshold potassium current. In contrast, Kv1.1 protein expression failed to increase in vitro, and changes in (Kv1.1-like) low threshold potassium current with time in culture were not significant. Electrophysiological recordings revealed that sodium inward currents increased with cultivation time. Thus, our data suggest that Kv3.1b expression occurs with the onset of functional synaptogenesis, while a different signal, absent from cultures of dissociated auditory brain stem, is needed for Kv1.1 expression. A biophysical model constructed with parameters from our recordings was used to investigate the functional impact of the currents mediated by these channels. We found that during development both high and low threshold potassium currents need to be increased in a concerted manner with the sodium conductance for the neurons to exhibit fast and phasic action potential firing and a narrow time window of coincidence detection.

Avian nucleus isthmi pars parvocellularis (Ipc) neurons are reciprocally connected with the layer 10 (L10) neurons in the optic tectum and respond with oscillatory bursts to visual stimulation. Our in vitro experiments show that both neuron types respond with regular spiking to somatic current injection and that the feedforward and feedback synaptic connections are excitatory, but of different strength and time course. To elucidate mechanisms of oscillatory bursting in this network of regularly spiking neurons, we investigated an experimentally constrained model of coupled leaky integrate-and-fire neurons with spike-rate adaptation. The model reproduces the observed Ipc oscillatory bursting in response to simulated visual stimulation. A scan through the model parameter volume reveals that Ipc oscillatory burst generation can be caused by strong and brief feedforward synaptic conductance changes. The mechanism is sensitive to the parameter values of spike-rate adaptation. In conclusion, we show that a network of regular-spiking neurons with feedforward excitation and spike-rate adaptation can generate oscillatory bursting in response to a constant input.

The magnetic compass of migratory birds has been suggested to be light-dependent. Retinal cryptochrome-expressing neurons and a forebrain region, "Cluster N", show high neuronal activity when night-migratory songbirds perform magnetic compass orientation. By combining neuronal tracing with behavioral experiments leading to sensory-driven gene expression of the neuronal activity marker ZENK during magnetic compass orientation, we demonstrate a functional neuronal connection between the retinal neurons and Cluster N via the visual thalamus. Thus, the two areas of the central nervous system being most active during magnetic compass orientation are part of an ascending visual processing stream, the thalamofugal pathway. Furthermore, Cluster N seems to be a specialized part of the visual wulst. These findings strongly support the hypothesis that migratory birds use their visual system to perceive the reference compass direction of the geomagnetic field and that migratory birds "see" the reference compass direction provided by the geomagnetic field.

The vertebrate nervous system has been shown to contain high concentrations of intracellular calcium-binding proteins, each of them with a restricted expression pattern in specific brain regions and specific neuronal subpopulations. Using immunohistochemical staining techniques, we analyzed the expression pattern of calbindin, calretinin and parvalbumin in visual brain areas of a songbird species, the zebra finch (Taeniopyga guttata). Here we show that the analyzed proteins are expressed in a complementary fashion within different brain substructures generally corresponding to functional subpathways of the avian visual system. In detail, calbindin is expressed in the brain structures that belong to the thalamofugal pathway, whereas parvalbumin-positive neurons are found in the brain structures that are part of the tectofugal visual pathway. Originally, the expression of calcium-binding proteins has been associated with specific morphological or neurochemical criteria of neurons. Our results suggest that their expression pattern also indicates a functional segregation of brain substructures linked to vision in the zebra finch brain. As the selective labeling of functional streams has also been shown for the visual system in mammalian species, function-selective expression of calcium-binding proteins might be a general feature of vertebrates.

Vitamin A is a major morphogen for the visual system. Most of its effects are mediated by retinoic acid (RA), whose developmental functions include pattern formation, neuronal differentiation and possibly axonal guidance. Although RA has been suggested to regulate development of the retina and its central projection, little is known about the distribution of retinoid receptors and binding proteins in the optic tectum, which in birds is the direct target of most retinofugal axons. We investigated the spatial and temporal distribution of the cellular retinoic acid binding protein-I (CRABP-I) in the chick midbrain. While the precise role of CRABP-I is still unknown, this is an intracellular transport protein for RA, which tends to be expressed in cells that are responsive to retinoic acid. Our data show immunoreactivity of CRABP-I in the tectal anlage at E2.5 and during the entire period of embryonic development. It was found in differentiating neurons of the generative zone, in migrating cells of the prospective stratum griseum et fibrosum superficiale and in mature neurons in this layer. In addition, we detected retinoid receptors RARalpha, RARbeta, RXRalpha, RXRbeta and RXRgamma in the developing tectum. Cell culture experiments demonstrate CRABP-I expression in a subpopulation of tectal neurons as they differentiate in vitro. These results are consistent with a regulatory role of RA in tectal neurogenesis and physiology.

The cholinergic division of the avian nucleus isthmi, the homolog of the mammalian nucleus parabigeminalis, is composed of the pars parvocellularis (Ipc) and pars semilunaris (SLu). Ipc and SLu were studied with in vivo and in vitro tracing and intracellular filling methods. 1) Both nuclei have reciprocal homotopic connections with the ipsilateral optic tectum. The SLu connection is more diffuse than that of Ipc. 2) Tectal inputs to Ipc and SLu are Brn3a-immunoreactive neurons in the inner sublayer of layer 10. Tectal neurons projecting on Ipc possess "shepherd's crook" axons and radial dendritic fields in layers 2-13. 3) Neurons in the mid-portion of Ipc possess a columnar spiny dendritic field. SLu neurons have a large, nonoriented spiny dendritic field. 4) Ipc terminals form a cylindrical brush-like arborization (35-50 microm wide) in layers 2-10, with extremely dense boutons in layers 3-6, and a diffuse arborization in layers 11-13. SLu neurons terminate in a wider column (120-180 microm wide) lacking the dust-like boutonal features of Ipc and extend in layers 4c-13 with dense arborizations in layers 4c, 6, and 9-13. 5) Ipc and SLu contain specialized fast potassium ion channels. We propose that dense arborizations of Ipc axons may be directed to the distal dendritic bottlebrushes of motion detecting tectal ganglion cells (TGCs). They may provide synchronous activation of a group of adjacent bottlebrushes of different TGCs of the same type via their intralaminar processes, and cross channel activation of different types of TGCs within the same column of visual space.

We studied the development of the projection from the external nucleus of the inferior colliculus (ICX) to the optic tectum (OT) in the barn owl. The projection was labeled by tracer application in vitro to either the OT or the ICX, or by staining ICX cells intracellularly with biocytin. The axons of ICX neurons bifurcated into an ascending branch that projected toward the OT and a descending branch that coursed caudally to an unknown target in the brainstem. Axons of the ICX were observed to grow into the OT from embryonic day 16 (E16) on. From E22 on, side branches of the axonal projections could be found within the OT. At the day of hatching (E32), the projection displayed a dorsoventral topography comparable to the adult owl; however, atopically projecting cells remained. The complexity of the axonal arborization in the adult barn owl was found to be slightly increased compared with the hatchling. The terminal area of individual ICX cells in the OT of the adult barn owl was still broad, a finding that had not been expected from the sharply defined physiological response properties of the bimodal neurons in the space map of the OT. However, the width of the termination zone was in accordance with the large dendritic tree of the adult ICX cells, because both spanned comparable angles in their respective maps. Our data suggest that a coarse projection from the ICX to the OT can develop without coherent sensory input and may, therefore, be innately determined.

Pit vipers (Crotalinae) have a specific sensory system that detects infrared radiation with bilateral pit organs in the upper jaw. Each pit organ consists of a thin membrane, innervated by three trigeminal nerve branches that project to a specific nucleus in the dorsal hindbrain. The known topographic organization of infrared signals in the optic tectum prompted us to test the implementation of spatiotopically aligned sensory maps through hierarchical neuronal levels from the peripheral epithelium to the first central site in the hindbrain, the nucleus of the lateral descending trigeminal tract (LTTD). The spatial organization of the anatomical connections was revealed in a novel in vitro whole-brain preparation of the western diamondback rattlesnake (Crotalus atrox) that allowed specific application of multiple neuronal tracers to identified pit-organ-supplying trigeminal nerve branches. After adequate survival times, the entire peripheral and central projections of fibers within the pit membrane and the LTTD became visible. This approach revealed a morphological partition of the pit membrane into three well-defined sensory areas with largely separated innervations by the three main branches. The peripheral segregation of infrared afferents in the sensory epithelium was matched by a differential termination of the afferents within different areas of the LTTD, with little overlap. This result demonstrates a topographic organizational principle of the snake infrared system that is implemented by maintaining spatially aligned representations of environmental infrared cues on the sensory epithelium through specific neuronal projections at the level of the first central processing stage, comparable to the visual system.

At least three identified cell types in the stratum griseum centrale (SGC) of the chick optic tectum mediate separate pathways from the retina to different subdivisions of the thalamic nucleus rotundus. Two of these, SGC type I and type II, constitute the major direct inputs to rotundal subdivisions that process various aspects of visual information, e.g., motion and luminance changes. Here, we examined the responses of these cell types to somatic current injection and synaptic input. We used a brain slice preparation of the chick tectum and applied whole-cell patch recordings, restricted electrical stimulation of dendritic endings, and subsequent labeling with biocytin. Type I neurons responded with regular sequences of bursts (“chattering”) to depolarizing current injection. Electrical stimulation of retinal afferents evoked a sharp-onset EPSP/burst response that was blocked with CNQX. The sharp-onset EPSP/burst response to synaptic stimulation persisted when the soma was hyperpolarized, thus suggesting the presence of dendritic spike generation. In contrast, the type II neurons responded to depolarizing current injection solely with an irregular sequence of individual spikes. Electrical stimulation of retinal afferents led to slow and long-lasting EPSPs that gave rise to one or several action potentials. In conclusion, the morphological distinct SGC type I and II neurons also have different response properties to retinal inputs. This difference is likely to have functional significance for the differential processing of visual information in the separate pathways from the retina to different subdivisions of the thalamic nucleus rotundus.

Many organisms use multimodal maps to generate coherent neuronal representations that allow adequate responses to stimuli that excite several sensory modalities. During ontogeny of these maps, one modality typically acts as the dominant system the other modalities are aligned to. A well studied model for the alignment of sensory maps is the calibration of the auditory space map by the visual system in the optic tectum of the barn owl. However, a projection from the optic tectum to the site of plasticity in the auditory pathway that could deliver an instructive signal has not been found so far. We have analyzed the development of the connectivity between the bimodal (visual and auditory) map of space in the barn owl’s optic tectum and the auditory space map in the inferior colliculus with tracing methods and intracellular fills. Neurons in the tectal stratum griseum centrale were found to be suited to deliver an alignment signal from the visual midbrain to the auditory pathway. These neurons are presumably part of the efferent tectal projection pathway that mediates head saccades. The implications of a sensory alignment signal possibly being delivered by a (pre)motor command pathway are discussed.

In avian and mammalian brains, visual information from the retina is conveyed to the telencephalon via two separate pathways: the thalamofugal and the tectofugal pathways. Recently, Karten et al. ([1997] J. Comp. Neurol. 387:449–465) examined a portion of the tectofugal pathway, the projection from the optic tectum to the nucleus rotundus thalami, in pigeons. They defined two distinct subpopulations of tectal neurons projecting from the stratum griseum centrale (SGC; tectal layer 13) to specific divisions of the rotundus. The goal of this study in chick was to verify the existence of the type I and type II SGC neurons, as defined by Karten et al., and then examine in greater detail the connectivity and morphology of these SGC neurons. Furthermore, our results suggest how the unique morphological characteristics of SGC neurons contribute to the large receptive fields (20–50°) found in physiological recordings and the SGC neuronal response to extremely small (ca. 0.05°), fast-moving (100°/second) stimuli. By injecting retrograde tracer into various divisions of the chick rotundus, we verified that, indeed, the chick did possess type I and type II SGC neurons, as well as a “new” type of SGC neuron, type III, that is not found in the pigeon. We then used intracellular cell-filling techniques to define further these three types of SGC neurons. Our examination revealed the following: Type I SGC neurons had large, circular dendritic fields (average diameter, 1,725 μm) composed of smooth dendrites and ending in spine-rich, bottlebrush endings located in retinorecipient tectal layer 5b; type II SGC neurons had elliptical dendritic fields (average 1,447 μm) and dendritic endings located never more superficially than tectal layer 8; and type III SGC neurons had large dendritic fields (average 1,800 μm) of unknown shape and bottlebrush dendritic endings located in retinorecipient tectal layer 4. We suggest that the neuronal features of the SGC neurons (i.e., bottlebrush dendritic endings and large dendritic fields) are key morphological characteristics for the detection of motion within the tectofugal pathway. Furthermore, because neurons with similar morphology have also been found in the tecta of both mammals and reptiles, we suggest that these neuronal features are fundamental components of a phylogenetically conserved system used for the “extrastriate” detection of motion in vertebrates. J. Comp. Neurol. 396:399–414, 1998. © 1998 Wiley-Liss, Inc.

The amphibian optic tectum and pretectum have been analyzed in detail anatomically and physiologically, and a specific model for tecto-pretectal interaction in the context of the visual guidance of behavior has been proposed. However, anatomical evidence for this model, particularly the precise pattern of pretecto-tectal connectivity, is lacking. Therefore, we stained pretectal neurons intracellularly in an in-vitro preparation of the salamanders Plethodon jordani and Hydromantes genei. Our results demonstrate that the projections of neurons of the nucleus praetectalis profundus are divergent and widespread. Individual neurons may project divergently to telencephalic (ipsilateral amygdala and striatum), diencephalic (ipsi-and contralateral thalamus, contralateral pretectum), and mesencephalic (ipsi- and contralateral tectum and tegmentum) centers, and to the ipsi- and contralateral medulla oblongata and rostral spinal cord. The projection of pretectal cells to the optic tectum is bilateral; axonal structures do not show discernible patterns and are present in all layers of the superficial white matter. A classification of pretectal neurons on the basis of axonal termination pattern or dendritic arborization has not been possible. Our results do not support the hypothesis that a distinct class of pretectal neurons projects to a particular subset of tectal cells. Rather, the pretectum appears to influence the tectum indirectly, acting either on retinal afferents or modulating inhibitory interneurons.

Interactions between pretectum and optic tectum of salamanders were analyzed by recording evoked potentials (EPs) in the optic tectum in vitro in response to stimulation of the contralateral optic nerve. Neither lesioning of the pretectum nor ablation of the medulla oblongata including the nucleus isthmi altered the shape of tectal EPs, suggesting that the tectal EP in this preparation reflects activation of tectal circuitry by retinal afferents without a major contribution from non-retinal afferents. To analyze the effect of stimulation of the pretectum on the tectal EP, we stimulated the pretectal area pharmacologically. Amplitudes of tectal EPs decreased rapidly after stimulation of the pretectum and recovered within minutes (glutamate) or hours (kainic acid). The pretectal influence on the tectal EP might act presynaptically on retinal afferents or by modulating the response of inhibitory interneurons.

To analyse cellular computation in the vertebrate brain, a thorough knowledge of the underlying anatomy, physiology and connectivity of the neuronal substrate is essential. This review compiles data on one of the best known structures of the vertebrate brain, the optic tectum of birds. The functions of this structure are multifold, but can be attributed largely to orientation and the basic analysis of sensory data in a spatial context. In the tectum, a wealth of data on physiology and anatomy has been gathered over more than a century and provides an excellent background for computational studies. The analysis of the optic tectum is facilitated by several principles of organisation, including the retinotopic input and the highly laminated layout with separated input and output layers. Moreover, the molecular mechanisms guiding the development and connectivity have been analysed in detail. As the avian tectum and the mammalian superior colliculus are partly homologous, the cellular mechanisms unraveled in the tectum can also be transferred to the colliculus and thus contribute to the understanding of the vertebrate visual system in general.