Avian Brain Research Articles

Local sleep in songbirds: different simultaneous sleep states across the avian pallium.

Wakefulness and sleep have often been treated as distinct and global brain states. However, an emerging body of evidence on the local regulation of sleep stages challenges this conventional view. Apart from unihemispheric sleep, the current data that support local variations of neural oscillations during sleep are focused on the homeostatic regulation of local sleep, i.e., the role preceding awake activity. Here, to examine local differences in brain activity during natural sleep, we recorded the electroencephalogram and the local field potential across multiple sites within the avian pallium of zebra finches without perturbing the previous awake state. We scored the sleep stages independently in each pallial site and found that the sleep stages are not pallium-wide phenomena but rather deviate widely across electrode sites. Importantly, deeper electrode sites had a dominant role in defining the temporal aspects of sleep state congruence. Altogether, these findings show that local regulation of sleep oscillations also occurs in the avian brain without prior awake recruitment of specific pallial circuits and in the absence of mammalian cortical neural architecture.

Functional MRI of imprinting memory in awake newborn domestic chicks

Filial imprinting, a crucial ethological paradigm, provides insights into the neurobiology of early learning and its long-term impact on behaviour. To date, invasive techniques like autoradiography or lesions have been used to study it, limiting the exploration of whole brain networks. Recent advances in fMRI for avian brains now open new windows to explore bird’s brain functions at the network level. We developed an fMRI technique for awake, newly hatched chicks, capturing BOLD signal changes during imprinting experiments. While early memory acquisition phases are understood, long-term storage and retrieval remain unclear. Our findings identified potential long-term storage of imprinting memories across a neural network, including the hippocampal formation, the medial striatum, the arcopallium, and the prefrontal-like nidopallium caudolaterale. This paradigm opens up new avenues for exploring the broader landscape of learning and memory in neonatal vertebrates, enhancing our understanding of behaviour and brain networks.

Development and Characterization of Primary Brain Cultures from Japanese Quail Embryos.

Cell cultures play a crucial role in neuroscience research, facilitating the elucidation of the complexities of cellular physiology and pathology. The relative simplicity in producing cultures and the accessibility to cells that the cultures provide, in contrast to in vivo settings, allow users to manipulate and monitor cells more easily at higher throughputs and lower costs. These are ideal for screening purposes and electrophysiological characterizations. Despite the prevalence of methodologies for producing brain cultures from various animal models, rodents in particular, approaches for culturing neurons (and glia) from birds are less established or completely absent as in the case of the Japanese quail model. Here, we present a unique culturing protocol for brain cells (e.g., neurons at different maturation levels, such as progenitor cells, excitatory and inhibitory neurons, microglia, and endothelial cells) from entire forebrains of Japanese quail embryos for high-throughput screening of viral vectors in vitro and other various purposes. Following dissection and digestion methods uniquely suited for avian brains, we tailored the growth media and culturing surface to allow the survival of quail brain cultures for more than three weeks in vitro. Key features • We introduce a detailed protocol for producing primary brain cultures from quail embryos' forebrains for up to 30 days. • We show that the cultures support in vitro viral transfections effectively. • We demonstrate the use of the cultures for rapid (days) screening for suitable viruses for quail brain cells, electrophysiological characterizations, and single mRNA sequencing.

Endocranial development in non-avian dinosaurs reveals an ontogenetic brain trajectory distinct from extant archosaurs

Modern birds possess highly encephalized brains that evolved from non-avian dinosaurs. Evolutionary shifts in developmental timing, namely juvenilization of adult phenotypes, have been proposed as a driver of head evolution along the dinosaur-bird transition, including brain morphology. Testing this hypothesis requires a sufficient developmental sampling of brain morphology in non-avian dinosaurs. In this study, we harness brain endocasts of a postnatal growth series of the ornithischian dinosaur Psittacosaurus and several other immature and mature non-avian dinosaurs to investigate how evolutionary changes to brain development are implicated in the origin of the avian brain. Using three-dimensional characterization of neuroanatomical shape across archosaurian reptiles, we demonstrate that (i) the brain of non-avian dinosaurs underwent a distinct developmental trajectory compared to alligators and crown birds; (ii) ornithischian and non-avialan theropod dinosaurs shared a similar developmental trajectory, suggesting that their derived trajectory evolved in their common ancestor; and (iii) the evolutionary shift in developmental trajectories is partly consistent with paedomorphosis underlying overall brain shape evolution along the dinosaur-bird transition; however, the heterochronic signal is not uniform across time and neuroanatomical region suggesting a highly mosaic acquisition of the avian brain form.

Flight patterns of monocularly occluded homing pigeons further reveal a left eye/right cerebral hemispheric advantage in the visual processing of familiar landscape/landmark features

Studying pigeons during homing has offered an opportunity to investigate visual–functional brain lateralization in the context of free flying and long distance navigation. In the current study we examine at a high scale of spatial resolution the flight paths of pigeons, which vary with respect to monocular and binocular treatment, as they familiarise themselves with landscape and landmark features during repeated flights home from two locations. The analysis of the flight paths of monocularly occluded pigeons revealed that pigeons using the right eye/left hemisphere visual system were more likely to display more tortuous paths over tracts of 500 m along the flight path home compared to both pigeons using the left eye/right hemisphere visual system and control birds. Accompanying this finding was the observation that pigeons using the right eye/left hemisphere system were more likely to perform flight “loops”, suggesting that, without right hemispheric visual processing, pigeons are more motivated to seek out visual input, behaviourally expanding the now monocularly reduced visual field. Taken together, the data are consistent with a more important role of the left eye/right hemisphere system in processing visual features of the landscape, which may contribute to the construction of the familiar landmark-based map used for navigation. More broadly, our data are consistent with the hypothesis that the right cerebral hemisphere of the avian brain plays a more important role in memorising and using the relational properties of visual stimuli to guide performance on spatial tasks.

Tutor auditory memory for guiding sensorimotor learning in birdsong.

Memory-guided motor shaping is necessary for sensorimotor learning. Vocal learning, such as speech development in human babies and song learning in bird juveniles, begins with the formation of an auditory template by hearing adult voices followed by vocally matching to the memorized template using auditory feedback. In zebra finches, the widely used songbird model system, only males develop individually unique stereotyped songs. The production of normal songs relies on auditory experience of tutor's songs (commonly their father's songs) during a critical period in development that consists of orchestrated auditory and sensorimotor phases. "Auditory templates" of tutor songs are thought to form in the brain to guide later vocal learning, while formation of "motor templates" of own song has been suggested to be necessary for the maintenance of stereotyped adult songs. Where these templates are formed in the brain and how they interact with other brain areas to guide song learning, presumably with template-matching error correction, remains to be clarified. Here, we review and discuss studies on auditory and motor templates in the avian brain. We suggest that distinct auditory and motor template systems exist that switch their functions during development.

A histological and diceCT-derived 3D reconstruction of the avian visual thalamofugal pathway

Amniotes feature two principal visual processing systems: the tectofugal and thalamofugal pathways. In most mammals, the thalamofugal pathway predominates, routing retinal afferents through the dorsolateral geniculate complex to the visual cortex. In most birds, the thalamofugal pathway often plays the lesser role with retinal afferents projecting to the principal optic thalami, a complex of several nuclei that resides in the dorsal thalamus. This thalamic complex sends projections to a forebrain structure called the Wulst, the terminus of the thalamofugal visual system. The thalamofugal pathway in birds serves many functions such as pattern discrimination, spatial memory, and navigation/migration. A comprehensive analysis of avian species has unveiled diverse subdivisions within the thalamic and forebrain structures, contingent on species, age, and techniques utilized. In this study, we documented the thalamofugal system in three dimensions by integrating histological and contrast-enhanced computed tomography imaging of the avian brain. Sections of two-week-old chick brains were cut in either coronal, sagittal, or horizontal planes and stained with Nissl and either Gallyas silver or Luxol Fast Blue. The thalamic principal optic complex and pallial Wulst were subdivided on the basis of cell and fiber density. Additionally, we utilized the technique of diffusible iodine-based contrast-enhanced computed tomography (diceCT) on a 5-week-old chick brain, and right eyeball. By merging diceCT data, stained histological sections, and information from the existing literature, a comprehensive three-dimensional model of the avian thalamofugal pathway was constructed. The use of a 3D model provides a clearer understanding of the structural and spatial organization of the thalamofugal system. The ability to integrate histochemical sections with diceCT 3D modeling is critical to better understanding the anatomical and physiologic organization of complex pathways such as the thalamofugal visual system.

Decoding Typical Flight States Based on Neural Signals from the Midbrain Motor Nuclei of Pigeons.

Exploring the neural encoding mechanism and decoding of motion state switching during flight can advance our knowledge of avian behavior control and contribute to the development of avian robots. However, limited acquisition equipment and neural signal quality have posed challenges, thus we understand little about the neural mechanisms of avian flight. We used chronically implanted micro-electrode arrays to record the local field potentials (LFPs) in the formation reticularis medialis mesencephali (FRM) of pigeons during various motion states in their natural outdoor flight. Subsequently, coherence-based functional connectivity networks under different bands were constructed and the topological features were extracted. Finally, we used a support vector machine model to decode different flight states. Our findings indicate that the gamma band (80-150 Hz) in the FRM exhibits significant power for identifying different states in pigeons. Specifically, the avian brain transmitted flight related information more efficiently during the accelerated take-off or decelerated landing states, compared with the uniform flight and baseline states. Finally, we achieved a best average accuracy of 0.86 using the connectivity features in the 80-150 Hz band and 0.89 using the fused features for state decoding. Our results open up possibilities for further research into the neural mechanism of avian flight and contribute to the understanding of flight behavior control in birds.

Brain surface morphology and ecological and macroevolutionary inferences of avian New World suboscines (Aves, Passeriformes, Tyrannides).

The New World suboscines (Passeriformes and Tyrannides) are one of the biggest endemic vertebrate radiations in South America, including the families Furnariidae and Tyrannidae. Avian brain morphology is a reliable proxy to study their evolution. The aim of this work is to elucidate whether the brains of these families reflect the ecological differences (e.g., feeding behavior) and to clarify macroevolutionary aspects of their neuroanatomy. Our hypotheses are as follows: Brain size is similar between both families and with other Passeriformes; brain morphology in Tyrannides is the result of the pressure of ecological factors; and brain disparity is low since they share ecological traits. Skulls of Furnariidae and Tyrannidae were micro-computed tomography-scanned, and three-dimensional models of the endocast were generated. Regression analyses were performed between brain volume and body mass. Linear and surface measurements were used to build phylomorphospaces and to calculate the amount of phylogenetic signal. Tyrannidae showed a larger brain disparity than Furnariidae, although it is not shaped by phylogeny in the Tyrannides. Furnariidae present enlarged Wulsts (eminentiae sagittales) but smaller optic lobes, while in Tyrannidae, it is the opposite. This could indicate that in Tyrannides there is a trade-off between the size of these two visual-related brain structures.

Three different seasonally expressed opsins are present in the brain of the Eared Dove, an opportunist breeder

Birds living at high latitudes perceive the photoperiod through deep-brain photoreceptors (DBP) located in deep-brain neurons. During long photoperiods the information transmitted by these photoreceptors increases the activity of the hypothalamic-pituitary-gonadal (HPG) axis, leading to gonadal development. The presence of photopigments such as VA-Opsin, Opn4, Opn5 and Opn2 in brain areas implicated in reproductive behaviors has been firmly established in several avian species with seasonal breeding, whereas their existence in opportunistic breeding birds remains unconfirmed. The Eared Dove is an urban and peri-urban dove that breeds throughout the year. Males of this species do not exhibit the typical gonadal regression/recrudescence cycle, thus posing the question of what occurs upstream of the HPG axis. We addressed this issue by first studying the presence of diverse opsins located in DBP in the brains of Eared Dove males and whether these photopigments changed their expression throughout the year. We carried out an immunohistochemistry analysis on three different opsins: Opn2 (rhodopsin), Opn3 and Opn5. Our results demonstrate the discrete neuroanatomical distribution of these opsins in the brain of Eared Dove males and strongly indicate different seasonal expressions. In the anterior region of the hypothalamus, Opn2-positive cells were detected throughout the year. By contrast, Opn5 was found to be strongly and seasonally expressed during winter in the anterior and the hypothalamic region. Opn3 was also found to be significantly and seasonally expressed during winter in the hypothalamic region. We thus demonstrate for the first time that males of the Eared Dove, have three different deep-brain opsin-expressing photoreceptors with differential location/distribution in the anterior and hypothalamic region and differential seasonality. The persistence of Opn2 and the strong seasonal expression of nonvisual photopigments Opn3 and Opn5 in two areas of the avian brain, which are associated with reproduction, could be the primary distinction between seasonal and opportunistic breeders.

Lateralisation, Hubs and Cognition in the Mammalian and Avian Brain

Lateralisation, Hubs and Cognition in the Mammalian and Avian Brain

Corvids optimize working memory by categorizing continuous stimuli

Working memory (WM) is a crucial element of the higher cognition of primates and corvid songbirds. Despite its importance, WM has a severely limited capacity and is vulnerable to noise. In primates, attractor dynamics mitigate the effect of noise by discretizing continuous information. Yet, it remains unclear whether similar dynamics are seen in avian brains. Here, we show jackdaws (Corvus monedula) have similar behavioral biases as humans; memories are less precise and more biased as memory demands increase. Model-based analysis reveal discrete attractors are evenly spread across the stimulus space. Altogether, our comparative approach suggests attractor dynamics in primates and corvids mitigate the effect of noise by systematically drifting towards specific attractors. By demonstrating this effect in an evolutionary distant species, our results strengthen attractor dynamics as general, adaptive biological principle to efficiently use WM.

Segmented spin-echo echo-planar imaging improves whole-brain BOLD functional MRI in awake pigeon brains.

Functional magnetic resonance imaging (fMRI) in awake small animals such as pigeons or songbirds opens a new window into the neural fundaments of cognitive behavior. However, high-field fMRI in the avian brain is challenging due to strong local magnetic field inhomogeneities caused by air cavities in the skull. A spoiled gradient-echo fMRI sequence has already been used to map the auditory network in songbirds, but due to susceptibility artifacts only 50% of the whole brain could be recorded. Since whole-brain fMRI coverage is vital to reveal whole-brain networks, an MRI sequence that is less susceptible to these artifacts was required. This was recently achieved in various bird species by using a rapid acquisition with relaxation enhancement (RARE) sequence. Weak blood oxygen level-dependent (BOLD) sensitivity, low temporal resolution, and heat caused by the long train of RF refocusing pulses are the main limits of RARE fMRI at high magnetic fields. To go beyond some of these limitations, we here describe the implementation of a two-segmented spin-echo echo-planar imaging (SE-EPI). The proposed sequence covers the whole brain of awake pigeons. The sequence was applied to investigate the auditory network in awake pigeons and assessed the relative merits of this method in comparison with the single-shot RARE sequence. At the same imaging resolution but with a volume acquisition of 3 s versus 4 s for RARE, the two-segmented SE-EPI provided twice the strength of BOLD activity compared with the single-shot RARE sequence, while the image signal-to-noise ratio (SNR) and in particular the temporal SNR were very similar for the two sequences. In addition, the activation patterns in two-segmented SE-EPI data are more symmetric and larger than single-shot RARE results. Two-segmented SE-EPI represents a valid alternative to the RARE sequence in avian fMRI research since it yields more than twice the BOLD sensitivity per unit of time with much less energy deposition and better temporal resolution, particularly for event-related experiments.

Molecular biology of serotonergic systems in avian brains.

Serotonin (5-hydroxytryptamine, 5-HT) is a phylogenetically conserved neurotransmitter and modulator. Neurons utilizing serotonin have been identified in the central nervous systems of all vertebrates. In the central serotonergic system of vertebrate species examined so far, serotonergic neurons have been confirmed to exist in clusters in the brainstem. Although many serotonin-regulated cognitive, behavioral, and emotional functions have been elucidated in mammals, equivalents remain poorly understood in non-mammalian vertebrates. The purpose of this review is to summarize current knowledge of the anatomical organization and molecular features of the avian central serotonergic system. In addition, selected key functions of serotonin are briefly reviewed. Gene association studies between serotonergic system related genes and behaviors in birds have elucidated that the serotonergic system is involved in the regulation of behavior in birds similar to that observed in mammals. The widespread distribution of serotonergic modulation in the central nervous system and the evolutionary conservation of the serotonergic system provide a strong foundation for understanding and comparing the evolutionary continuity of neural circuits controlling corresponding brain functions within vertebrates. The main focus of this review is the chicken brain, with this type of poultry used as a model bird. The chicken is widely used not only as a model for answering questions in developmental biology and as a model for agriculturally useful breeding, but also in research relating to cognitive, behavioral, and emotional processes. In addition to a wealth of prior research on the projection relationships of avian brain regions, detailed subdivision similarities between avian and mammalian brains have recently been identified. Therefore, identifying the neural circuits modulated by the serotonergic system in avian brains may provide an interesting opportunity for detailed comparative studies of the function of serotonergic systems in mammals.

Dim artificial light at night alters immediate early gene expression throughout the avian brain.

Artificial light at night (ALAN) is a pervasive pollutant that alters physiology and behavior. However, the underlying mechanisms triggering these alterations are unknown, as previous work shows that dim levels of ALAN may have a masking effect, bypassing the central clock. Light stimulates neuronal activity in numerous brain regions which could in turn activate downstream effectors regulating physiological response. In the present study, taking advantage of immediate early gene (IEG) expression as a proxy for neuronal activity, we determined the brain regions activated in response to ALAN. We exposed zebra finches to dim ALAN (1.5 lux) and analyzed 24 regions throughout the brain. We found that the overall expression of two different IEGs, cFos and ZENK, in birds exposed to ALAN were significantly different from birds inactive at night. Additionally, we found that ALAN-exposed birds had significantly different IEG expression from birds inactive at night and active during the day in several brain areas associated with vision, movement, learning and memory, pain processing, and hormone regulation. These results give insight into the mechanistic pathways responding to ALAN that underlie downstream, well-documented behavioral and physiological changes.

Nest complexity correlates with larger brain size but smaller body mass across bird species.

Amniotes differ substantially in absolute and relative brain size after controlling for allometry, and numerous hypotheses have been proposed to explain brain size evolution. Brain size is thought to correlate with processing capacity and the brain's ability to support complex manipulation such as nest-building skills. The increased complexity of nest structure is supposed to be a measure of an ability to manipulate nesting material into the required shape. The degree of nest-structure complexity is also supposed to be associated with body mass, partly because small species lose heat faster and delicate and insulated nests are more crucial for temperature control of eggs during incubation by small birds. Here, we conducted comparative analyses to test these hypotheses by investigating whether the complexity of species-typical nest structure can be explained by brain size and body mass (a covariate also to control for allometric effects on brain size) across 1353 bird species from 147 families. Consistent with these hypotheses, our results revealed that avian brain size increases as the complexity of the nest structure increases after controlling for a significant effect of body size, and also that a negative relationship exists between nest complexity and body mass.

Brain Gross Anatomy and Cerebellar Histology of the Cattle Egret (Bubulcus ibis)

Abstract The structural parts of brain are similar in all vertebrates, but they differ in their complexity and organization. The avian brain for instance, is organised differently compared to mammals, with variations existing in the relative size and location of their structures. The cattle egret (Bubulcus ibis) is a cosmopolitan avian species native to Africa. Despite their economic importance as excellent sentinels in assessing environmental pollutants, there is a dearth of information on their neuroanatomy. Hence, we here investigated their gross morphological features and morphometric parameters, and the cerebellar histology. Twelve transcardi-ally perfused juvenile cattle egrets were studied, with the body and brain weights and brain linear measurements considered. The brain weight constituted 0.46 % of the total body mass, with a strong positive correlation recorded between the body and brain weights (r = 0.9204). Morphologically, the brain was lissencephalic, with the corpus callosum absent and the olfactory bulbs rudimentary with no discernible division into the olfactory bulb, olfactory tract and olfactory lobe. We observed prominent sagittal eminence and vallecula telencephali, as well as an obvious fovea limbica on the dorsal and lateral surfaces of the cerebral hemispheres, respectively. The diencephalic structures were completely covered by telencephalon, with the pineal gland occupying the polygonal space between cerebrum, optic lobe and cerebellum. The mesencephalic tectum appeared as a large oval bilaterally bulging structure with prominent optic tracts and constituted the bulk of the midbrain. There was a dorsal extension of the fourth ventricle into the cerebellum known as the ventriculus cerebelli and the cerebellar histology presented a persistent external granular layering suggestive of a potential for adult neurogenesis. Our data has added relevant literature on the cattle egret brain and could prove useful in comparative, developmental and evolutionary avian neuroanatomy.

Comparative genomics of two Empidonax flycatchers reveal candidate genes for bird song production.

Whole-genome level comparisons of sister taxa that vary in phenotype against a background of high genomic similarity can be used to identify the genomic regions that might underlie their phenotypic differences. In wild birds, this exploratory approach has detected markers associated with plumage coloration, beak and wing morphology, and complex behavioral traits like migration. Here, we use genomic comparisons of two closely related suboscine flycatchers (Empidonax difficilis and E. occidentalis) and their hybrids to search for candidate genes underlying their variation in innate vocal signals. We sequenced the genomes of 20 flycatchers that sang one of two species-specific parental song types and 14 putative hybrid individuals with intermediate song types. In the resulting genomic comparisons, we found six areas of high differentiation that may be associated with variation in non-learned songs. These narrow regions of genomic differentiation contain a total of 67 described genes, three of which have been previously associated with forms of language impairment and dyslexia in humans, and 18 of which are known to be differentially expressed in the song nuclei regions of the avian brain compared to adjacent parts of the avian brain. This 'natural experiment' therefore may help identify loci associated with song differences that merit further study across bird lineages with both learned and innate vocalizations.

Multi-channel recordings reveal age-related differences in the sleep of juvenile and adult zebra finches

Despite their phylogenetic differences and distinct pallial structures, mammals and birds show similar electroencephalography (EEG) traces during sleep, consisting of distinct rapid eye movement (REM) sleep and slow wave sleep (SWS) stages. Studies in human and a limited number of other mammalian species show that this organization of sleep into interleaving stages undergoes radical changes during lifetime. Do these age-dependent variations in sleep patterns also occur in the avian brain? Does vocal learning have an effect on sleep patterns in birds? To answer these questions, we recorded multi-channel sleep EEG from juvenile and adult zebra finches for several nights. Whereas adults spent more time in SWS and REM sleep, juveniles spent more time in intermediate sleep (IS). The amount of IS was significantly larger in male juveniles engaged in vocal learning compared to female juveniles, which suggests that IS could be important for vocal learning. In addition, we observed that functional connectivity increased rapidly during maturation of young juveniles, and was stable or declined at older ages. Synchronous activity during sleep was larger for recording sites in the left hemisphere for both juveniles and adults, and generally intra-hemispheric synchrony was larger than inter-hemispheric synchrony during sleep. A graph theory analysis revealed that in adults, highly correlated EEG activity tended to be distributed across fewer networks that were spread across a wider area of the brain, whereas in juveniles, highly correlated EEG activity was distributed across more numerous, albeit smaller, networks in the brain. Overall, our results reveal that significant changes occur in the neural signatures of sleep during maturation in an avian brain.

(New neurons) singing in the avian brain.

(New neurons) singing in the avian brain.