Crows "count" the number of self-generated vocalizations.

Producing a specific number of vocalizations with purpose requires a sophisticated combination of numerical abilities and vocal control. Whether this capacity exists in animals other than humans is yet unknown. We show that crows can flexibly produce variable numbers of one to four vocalizations in response to arbitrary cues associated with numerical values. The acoustic features of the first vocalization of a sequence were predictive of the total number of vocalizations, indicating a planning process. Moreover, the acoustic features of vocal units predicted their order in the sequence and could be used to read out counting errors during vocal production.

Lesions in a songbird vocal circuit increase variability in song syntax.

Complex skills like speech and dance are composed of ordered sequences of simpler elements, but the neuronal basis for the syntactic ordering of actions is poorly understood. Birdsong is a learned vocal behavior composed of syntactically ordered syllables, controlled in part by the songbird premotor nucleus HVC (proper name). Here, we test whether one of HVC's recurrent inputs, mMAN (medial magnocellular nucleus of the anterior nidopallium), contributes to sequencing in adult male Bengalese finches (Lonchura striata domestica). Bengalese finch song includes several patterns: (1) chunks, comprising stereotyped syllable sequences; (2) branch points, where a given syllable can be followed probabilistically by multiple syllables; and (3) repeat phrases, where individual syllables are repeated variable numbers of times. We found that following bilateral lesions of mMAN, acoustic structure of syllables remained largely intact, but sequencing became more variable, as evidenced by 'breaks' in previously stereotyped chunks, increased uncertainty at branch points, and increased variability in repeat numbers. Our results show that mMAN contributes to the variable sequencing of vocal elements in Bengalese finch song and demonstrate the influence of recurrent projections to HVC. Furthermore, they highlight the utility of species with complex syntax in investigating neuronal control of ordered sequences.

Crows protect visual working memory against interference.

Working memory, the ability to actively maintain and manipulate information across time, is key to intelligent behavior. Because of the limited capacity of working memory, relevant information needs to be protected against distracting representations. Whether birds can resist distractors and safeguard memorized relevant information is unclear. We trained carrion crows in a delayed match-to-sample task to memorize an image while resisting other, interfering stimuli. We found that the repetition of the sample stimulus during the memory delay improved performance accuracy and accelerated reaction time relative to a reference condition with a neutral interfering stimulus. In contrast, the presentation of the image that constituted the subsequent non-match test stimulus mildly weakened performance. However, the crows' robust performance in this most demanding distractor condition indicates that sample information was actively protected from being overwritten by the distractor. These data show that crows can cognitively control and safeguard behaviorally relevant working memory contents.

Songbirds can learn flexible contextual control over syllable sequencing.

The flexible control of sequential behavior is a fundamental aspect of speech, enabling endless reordering of a limited set of learned vocal elements (syllables or words). Songbirds are phylogenetically distant from humans but share both the capacity for vocal learning and neural circuitry for vocal control that includes direct pallial-brainstem projections. Based on these similarities, we hypothesized that songbirds might likewise be able to learn flexible, moment-by-moment control over vocalizations. Here, we demonstrate that Bengalese finches (Lonchura striata domestica), which sing variable syllable sequences, can learn to rapidly modify the probability of specific sequences (e.g. 'ab-c' versus 'ab-d') in response to arbitrary visual cues. Moreover, once learned, this modulation of sequencing occurs immediately following changes in contextual cues and persists without external reinforcement. Our findings reveal a capacity in songbirds for learned contextual control over syllable sequencing that parallels human cognitive control over syllable sequencing in speech.

Human speech and birdsong share numerous parallels. Both humans and birds learn their vocalizations during critical phases early in life, and both learn by imitating adults. Moreover, both humans and songbirds possess specific circuits in the brain that connect the forebrain to midbrain vocal centers. Humans can flexibly control what they say and how by reordering a fixed set of syllables into endless combinations, an ability critical to human speech and language. Birdsongs also vary depending on their context, and melodies to seduce a mate will be different from aggressive songs to warn other males to stay away. However, so far it was unclear whether songbirds are also capable of modifying songs independent of social or other naturally relevant contexts. To test whether birds can control their songs in a purposeful way, Veit et al. trained adult male Bengalese finches to change the sequence of their songs in response to random colored lights that had no natural meaning to the birds. A specific computer program was used to detect different variations on a theme that the bird naturally produced (for example, "ab-c" versus "ab-d"), and rewarded birds for singing one sequence when the light was yellow, and the other when it was green. Gradually, the finches learned to modify their songs and were able to switch between the appropriate sequences as soon as the light cues changed. This ability persisted for days, even without any further training. This suggests that songbirds can learn to flexibly and purposefully modify the way in which they sequence the notes in their songs, in a manner that parallels how humans control syllable sequencing in speech. Moreover, birds can learn to do this 'on command' in response to an arbitrarily chosen signal, even if it is not something that would impact their song in nature. Songbirds are an important model to study brain circuits involved in vocal learning. They are one of the few animals that, like humans, learn their vocalizations by imitating conspecifics. The finding that they can also flexibly control vocalizations may help shed light on the interactions between cognitive processing and sophisticated vocal learning abilities.

Neurons in the crow nidopallium caudolaterale encode varying durations of visual working memory periods.

Adaptive sequential behaviors rely on the bridging and integration of temporally separate information for the realization of prospective goals. Corvids' remarkable behavioral flexibility is thought to depend on the workings of the nidopallium caudolaterale (NCL), a high-level avian associative forebrain area. We trained carrion crows to remember visual items for three alternating delay durations in a delayed match-to-sample task and recorded single-unit activity from the NCL. Sample-selective delay activity, a correlate of visual working memory, was maintained throughout different working memory durations. Delay responses remained selective for the same preferred sample item across blocks with different delay durations. However, selectivity strength decreased with increasing delay durations, mirroring worsened behavioral performance with longer memory delays. Behavioral relevance of delay activity was further evidenced by reduced encoding of the preferred sample item during error trials. In addition, NCL neurons adapted their time-dependent discharges to blocks of different memory durations, so that delay duration could be successfully classified based on population activity a few trials after the delay duration switched. Therefore, NCL neurons not only maintain information from individual trials, but also keep track of the duration for which this information is needed in the context of the task. These results strengthen the role of corvid NCL in maintaining working memory for flexible control of temporally extended goal-directed behavior.

Learning Recruits Neurons Representing Previously Established Associations in the Corvid Endbrain.

Crows quickly learn arbitrary associations. As a neuronal correlate of this behavior, single neurons in the corvid endbrain area nidopallium caudolaterale (NCL) change their response properties during association learning. In crows performing a delayed association task that required them to map both familiar and novel sample pictures to the same two choice pictures, NCL neurons established a common, prospective code for associations. Here, we report that neuronal tuning changes during learning were not distributed equally in the recorded population of NCL neurons. Instead, such learning-related changes relied almost exclusively on neurons which were already encoding familiar associations. Only in such neurons did behavioral improvements during learning of novel associations coincide with increasing selectivity over the learning process. The size and direction of selectivity for familiar and newly learned associations were highly correlated. These increases in selectivity for novel associations occurred only late in the delay period. Moreover, NCL neurons discriminated correct from erroneous trial outcome based on feedback signals at the end of the trial, particularly in newly learned associations. Our results indicate that task-relevant changes during association learning are not distributed within the population of corvid NCL neurons but rather are restricted to a specific group of association-selective neurons. Such association neurons in the multimodal cognitive integration area NCL likely play an important role during highly flexible behavior in corvids.

Spatially Tuned Neurons in Corvid Nidopallium Caudolaterale Signal Target Position During Visual Search.

The avian pallial endbrain area nidopallium caudolaterale (NCL) shows important similarities to mammalian prefrontal cortex in connectivity, dopamine neurochemistry, and function. Neuronal processing in NCL has been studied with respect to sensory, cognitive, and reward information, but little is known about its role in more direct control of motor behavior. We investigated NCL activity during the choice period of a delayed match-to-sample task, as 2 trained crows searched and selected a previously remembered visual target among an array of 4 pictures. The crows exhibited behavioral response patterns consistent with serial visual search. Many single NCL neurons were spatially tuned to specific target positions during visual search and directed motor behavior. Moreover, single NCL neurons dynamically changed their tuning properties to represent different behaviorally relevant task variables across the trial. In consecutive task periods, single neurons responded to visual stimuli, stored stimulus information in working memory, guided goal-directed behavior depending on the remembered target picture, and encoded trial outcomes. This flexible encoding of all task-relevant aspects in the executive control of goal-directed behavior represents a striking convergence to neuronal encoding in primate prefrontal cortex. These data highlight key properties of associative endbrain areas underlying flexible cognitive behavior in corvids and primates.

Associative learning rapidly establishes neuronal representations of upcoming behavioral choices in crows.

The ability to form associations between behaviorally relevant sensory stimuli is fundamental for goal-directed behaviors. We investigated neuronal activity in the telencephalic area nidopallium caudolaterale (NCL) while two crows (Corvus corone) performed a delayed association task. Whereas some paired associates were familiar to the crows, novel associations had to be learned and mapped to the same target stimuli within a single session. We found neurons that prospectively encoded the chosen test item during the delay for both familiar and newly learned associations. These neurons increased their selectivity during learning in parallel with the crows' increased behavioral performance. Thus, sustained activity in the NCL actively processes information for the upcoming behavioral choice. These data provide new insights into memory representations of behaviorally meaningful stimuli in birds, and how such representations are formed during learning. The findings suggest that the NCL plays a role in learning arbitrary associations, a cornerstone of corvids' remarkable behavioral flexibility and adaptability.

Neuronal correlates of visual working memory in the corvid endbrain.

The concept of working memory is key to cognitive functioning. Working memory encompasses the capacity to retain immediately past information, to process this information, and to use it to guide goal-directed behavior. Corvid songbirds are renowned for their high-level cognitive capabilities, but where and how visual information is temporarily retained by neurons in the avian brain in a behaviorally relevant way remains poorly understood. We trained four carrion crows (Corvus corone) on versions of a delayed match-to-sample task that required the crows to remember a visual stimulus for later comparison. While the crows performed the task, we recorded the activity of single neurons in the nidopallium caudolaterale (NCL), a pallial association area of the avian endbrain. We show that many NCL neurons encode information about visual stimuli and temporarily maintain this information after the stimulus disappeared by sustained delay activity. Selective delay activity allows the birds to hold relevant information in memory and correlates with discrimination behavior. This suggests that sustained activity of NCL neurons is a neuronal correlate of visual working memory in the corvid brain and serves to bridge temporal gaps, thereby offering a workspace for processing immediately past visual information.

Abstract rule neurons in the endbrain support intelligent behaviour in corvid songbirds.

Despite the lack of a layered neocortex and fundamental differences in endbrain organization in birds compared with mammals, intelligent species evolved from both vertebrate classes. Among birds, corvids show exceptional cognitive flexibility. Here we explore the neuronal foundation of corvid cognition by recording single-unit activity from an association area known as the nidopallium caudolaterale (NCL) while carrion crows make flexible rule-guided decisions, a hallmark of executive control functions. The most prevalent activity in NCL represents the behavioural rules, while abstracting over sample images and sensory modalities of the rule cues. Rule coding is weaker in error trials, thus predicting the crows' behavioural decisions. This suggests that the abstraction of general principles may be an important function of the NCL, mirroring the function of primate prefrontal cortex. These findings emphasize that intelligence in vertebrates does not necessarily rely on a neocortex but can be realized in endbrain circuitries that developed independently via convergent evolution.

Two distinct modes of forebrain circuit dynamics underlie temporal patterning in the vocalizations of young songbirds.

Accurate timing is a critical aspect of motor control, yet the temporal structure of many mature behaviors emerges during learning from highly variable exploratory actions. How does a developing brain acquire the precise control of timing in behavioral sequences? To investigate the development of timing, we analyzed the songs of young juvenile zebra finches. These highly variable vocalizations, akin to human babbling, gradually develop into temporally stereotyped adult songs. We find that the durations of syllables and silences in juvenile singing are formed by a mixture of two distinct modes of timing: a random mode producing broadly distributed durations early in development, and a stereotyped mode underlying the gradual emergence of stereotyped durations. Using lesions, inactivations, and localized brain cooling, we investigated the roles of neural dynamics within two premotor cortical areas in the production of these temporal modes. We find that LMAN (lateral magnocellular nucleus of the nidopallium) is required specifically for the generation of the random mode of timing and that mild cooling of LMAN causes an increase in the durations produced by this mode. On the contrary, HVC (used as a proper name) is required specifically for producing the stereotyped mode of timing, and its cooling causes a slowing of all stereotyped components. These results show that two neural pathways contribute to the timing of juvenile songs and suggest an interesting organization in the forebrain, whereby different brain areas are specialized for the production of distinct forms of neural dynamics.

Learning to breathe and sing: development of respiratory-vocal coordination in young songbirds.

How do animals with learned vocalizations coordinate vocal production with respiration? Songbirds such as the zebra finch learn their songs, beginning with highly variable babbling vocalizations known as subsong. After several weeks of practice, zebra finches are able to produce a precisely timed pattern of syllables and silences, precisely coordinated with expiratory and inspiratory pulses (Franz M, Goller F. J Neurobiol 51: 129-141, 2002). While respiration in adult song is well described, relatively little is known about respiratory patterns in subsong or about the processes by which respiratory and vocal patterns become coordinated. To address these questions, we recorded thoracic air sac pressure in juvenile zebra finches prior to the appearance of any consistent temporal or acoustic structure in their songs. We found that subsong contains brief inspiratory pulses (50 ms) alternating with longer pulses of sustained expiratory pressure (50-500 ms). In striking contrast to adult song, expiratory pulses often contained multiple (0-8) variably timed syllables separated by expiratory gaps and were only partially vocalized. During development, expiratory pulses became shorter and more stereotyped in duration with shorter and fewer nonvocalized parts. These developmental changes eventually resulted in the production of a single syllable per expiratory pulse and a single inspiratory pulse filling each gap, forming a coordinated sequence similar to that of adult song. To examine the role of forebrain song-control nuclei in the development of respiratory patterns, we performed pressure recordings before and after lesions of nucleus HVC (proper name) and found that this manipulation reverses the developmental trends in measures of the respiratory pattern.