Developmental polychlorinated biphenyl exposure influences adult zebra finch reproductive behaviour.

Polychlorinated biphenyls (PCBs) are worldwide chemical pollutants that have been linked to disrupted reproduction and altered sexual behaviour in many organisms. However, the effect of developmental PCB-exposure on adult passerine reproductive behaviour remains unknown. A commercial PCB mixture (Aroclor 1242) or an estrogenic congener (PCB 52) were administered in sublethal amounts to nestling zebra finches (Taeniopygia guttata) in the laboratory to identify effects of developmental PCB-exposure on adult zebra finch reproductive parameters. Results indicate that although traditional measures of reproductive success are not altered by this PCB dosage, PCBs do alter sexual behaviours such as male song and nesting behaviour. Males treated with PCB 52 in the nest sang significantly fewer syllables than control males, while females treated with Aroclor 1242 in the nest showed the strongest song preferences. PCB treatment also caused an increase in the number of nesting attempts and abandoned nests in the Aroclor 1242 treatment relative to the PCB 52 treatment, and offspring with control fathers fledged significantly earlier than those with fathers treated with Aroclor 1242. Behavioural differences between males seem to best explain these reproductive effects, most notably aggression. These findings suggest that sublethal PCB-exposure during development can significantly alter key reproductive characteristics of adult zebra finches, likely reducing fitness in the wild.

Concerted and mosaic evolution of functional modules in songbird brains.

Vertebrate brains differ in overall size, composition and functional capacities, but the evolutionary processes linking these traits are unclear. Two leading models offer opposing views: the concerted model ascribes major dimensions of covariation in brain structures to developmental events, whereas the mosaic model relates divergent structures to functional capabilities. The models are often cast as incompatible, but they must be unified to explain how adaptive changes in brain structure arise from pre-existing architectures and developmental mechanisms. Here we show that variation in the sizes of discrete neural systems in songbirds, a species-rich group exhibiting diverse behavioural and ecological specializations, supports major elements of both models. In accordance with the concerted model, most variation in nucleus volumes is shared across functional domains and allometry is related to developmental sequence. Per the mosaic model, residual variation in nucleus volumes is correlated within functional systems and predicts specific behavioural capabilities. These comparisons indicate that oscine brains evolved primarily as a coordinated whole but also experienced significant, independent modifications to dedicated systems from specific selection pressures. Finally, patterns of covariation between species and brain areas hint at underlying developmental mechanisms.

Juvenile zebra finches learn the underlying structural regularities of their fathers' song.

Natural behaviors, such as foraging, tool use, social interaction, birdsong, and language, exhibit branching sequential structure. Such structure should be learnable if it can be inferred from the statistics of early experience. We report that juvenile zebra finches learn such sequential structure in song. Song learning in finches has been extensively studied, and it is generally believed that young males acquire song by imitating tutors (Zann, 1996). Variability in the order of elements in an individual's mature song occurs, but the degree to which variation in a zebra finch's song follows statistical regularities has not been quantified, as it has typically been dismissed as production error (Sturdy et al., 1999). Allowing for the possibility that such variation in song is non-random and learnable, we applied a novel analytical approach, based on graph-structured finite-state grammars, to each individual's full corpus of renditions of songs. This method does not assume syllable-level correspondence between individuals. We find that song variation can be described by probabilistic finite-state graph grammars that are individually distinct, and that the graphs of juveniles are more similar to those of their fathers than to those of other adult males. This grammatical learning is a new parallel between birdsong and language. Our method can be applied across species and contexts to analyze complex variable learned behaviors, as distinct as foraging, tool use, and language.

The effect of polychlorinated biphenyls on the song of two passerine species.

Polychlorinated biphenyls (PCBs) are synthetic chemical pollutants with demonstrated detrimental toxic and developmental effects on humans and wildlife. Laboratory studies suggest that PCBs influence behavior due to their effects on endocrine and neurological systems, yet little is known about the behavioral consequences of sublethal PCB exposure in the field. Additionally, specific PCB congener data (in contrast to total PCB load) is necessary to understand the possible effects of PCBs in living organisms since number and position of chlorine substitution in a PCB molecule dictates the toxicity and chemical fate of individual PCB congeners. We non-lethally investigated total PCB loads, congener specific PCB profiles, and songs of black-capped chickadees (Poecile atricapillus) and song sparrows (Melospiza melodia) along a historical PCB gradient at the Hudson River in New York State. Our results indicate that black-capped chickadees and song sparrows have higher total blood PCBs in regions with higher historic PCB contamination. The two bird species varied substantially in their congener-specific PCB profiles; within sites, song sparrows showed a significantly higher proportion of lower chlorinated PCBs, while black-capped chickadees had higher proportions of highly chlorinated PCBs. In areas of PCB pollution, the species-specific identity signal in black-capped chickadee song varied significantly, while variation in song sparrow trill performance was best predicted by the mono-ortho PCB load. Thus, PCBs may affect song production, an important component of communication in birds. In conclusion, we suggest that the ramifications of changes in song quality for bird populations may extend the toxic effects of environmental PCB pollution.

Motor pathway convergence predicts syllable repertoire size in oscine birds.

Behavioral specializations are frequently associated with expansions of the brain regions controlling them. This principle of proper mass spans sensory, motor, and cognitive abilities and has been observed in a wide variety of vertebrate species. Yet, it is unknown if this concept extrapolates to entire neural pathways or how selection on a behavioral capacity might otherwise shape circuit structure. We investigate these questions by comparing the songs and neuroanatomy of 49 species from 17 families of songbirds, which vary immensely in the number of unique song components they produce and possess a conserved neural network dedicated to this behavior. We find that syllable repertoire size is strongly related to the degree of song motor pathway convergence. Repertoire size is more accurately predicted by the number of neurons in higher motor areas relative to that in their downstream targets than by the overall number of neurons in the song motor pathway. Additionally, the convergence values along serial premotor and primary motor projections account for distinct portions of the behavioral variation. These findings suggest that selection on song has independently shaped different components of this hierarchical pathway, and they elucidate how changes in pathway structure could have underlain elaborations of this learned motor behavior.

Captivity reduces hippocampal volume but not survival of new cells in a food-storing bird.

In many naturalistic studies of the hippocampus wild animals are held in captivity. To test if captivity itself affects hippocampal integrity, adult black-capped chickadees (Poecile atricapilla) were caught in the fall, injected with bromodeoxyuridine to mark neurogenesis, and alternately released to the wild or held in captivity. The wild birds were recaptured after 4-6 weeks and perfused simultaneously with their captive counterparts. The hippocampus of captive birds was 23% smaller than wild birds, with no hemispheric differences in volume within groups. Between groups there was no statistically significant difference in the size of the telencephalon, or in the number and density of surviving new cells. Proximate causes of the reduced hippocampal volume could include stress, lack of exercise, diminished social interaction, or limited caching opportunity-a hippocampal-dependent activity. The results suggest the avian hippocampus-a structure essential for rapid, complex relational and spatial learning-is both plastic and sensitive, much as in mammals, including humans.

Development of a brain nucleus involved in song production in zebra finches (Taeniopygia guttata) is disrupted by Aroclor 1248.

We studied whether polychlorinated biphenyls (PCBs) may alter the development of song control brain nuclei in zebra finch (Taeniopygia guttata) offspring of pulse-exposed hens. We orally administered 40 microg of Aroclor 1248 to adult female finches before egg laying. When the progeny were 50 d old, we measured the volumes of the song control nuclei robustus arcopallialis (RA) and higher vocal center (HVC) using light microscopy. Both male and female progeny of exposed birds had a significantly smaller RA than control birds (36 and 16%, respectively; p < or = 0.05). The HVC did not differ in either sex between exposed and control groups. Perhaps impaired development of RA was caused by PCB action on steroid receptors. We conclude that animals living in contaminated areas may be at risk of neurological damage in hormone-sensitive brain areas and that changes in brain nuclei related to song may be a sensitive indicator of low-level PCB exposure.

Immediate early gene (ZENK) responses to song in juvenile female and male zebra finches: effects of rearing environment.

Accurate song perception is likely to be as important for female songbirds as it is for male songbirds. Male zebra finches (Taeniopygia guttata) show differential ZENK expression to conspecific and heterospecific songs by day 30 posthatch in auditory perceptual brain regions such as the caudomedial nidopallium (NCM) and the caudomedial mesopallium (CMM). The current study examined ZENK expression in response to songs of different qualities at day 45 posthatch in both sexes. Normally reared juvenile zebra finches showed higher densities of immunopositive nuclei in both the dorsal and ventral areas of NCM and CMM (formerly cmHV), but not HA, a visual area, in response to normal song over untutored song or silence. Male and female patterns of ZENK expression did not differ. We next compared responses of birds reared without exposure to normal song (untutored) to those of normally reared birds. Untutored birds did not show higher responses to normal song than to untutored song in the three song perception areas. Furthermore, untutored birds of both sexes showed lower densities of immunopositive nuclei in all four areas than did normally reared birds. In addition, ZENK expression was greater in untutored females than in males in the dorsal portion of NCM and in CMM. Our findings suggest that at least some neural mechanisms of song perception are in place in socially reared female and male finches at an early age. Furthermore, early exposure to song tutors affects responses to song stimuli.

Adult female and male zebra finches show distinct patterns of spine deficits in an auditory area and in the song system when reared without exposure to normal adult song.

Male songbirds typically require exposure to normal adult conspecific song during development in order to learn a normal song of their own. Females require exposure to conspecific song during development in order to select high-quality, learned song over the incomplete song produced by males reared in isolation. Altering males' opportunity for song learning during development affects the neuroanatomy of brain regions involved in song production (the song system), but in females the neural effects of song learning are unknown. We raised male and female zebra finches (Taeniopygia guttata) with differing amounts of exposure to singing males during development. At 120 days, we Golgi-stained their brains and measured the frequency of dendritic spines in brain areas used in song perception or production. We found that females reared with little or no exposure to song have 31% fewer dendritic spines per unit length of dendrite in caudomedial nidopallium (NCM), a brain area activated by song perception, compared to control females. The deprived females had small deficits in the frequency of spines in HVC, a region activated by song production in males. Males with limited exposure to song had a 24% lower spine density in HVC than controls but only a 10% lower density in NCM. These data support the hypothesis that NCM is important in auditory learning, while HVC is involved in sensorimotor learning, and that these capacities are differentially emphasized in the two sexes.

Reproductive impairment in zebra finches (Taeniopygia guttata).

The effects of polychlorinated biphenyls (PCBs) as compounds that may disrupt endocrine activity and, consequently, alter reproductive performance were investigated in altricial zebra finches (Taeniopygia guttata). The breeding performance and breeding cycle of zebra finches differed significantly between nonexposed birds and those experimentally pulse-exposed to Aroclor 1248, a PCB compound (40 microg/bird). Aroclor-exposed birds showed significantly increased numbers of clutches laid, nests constructed per pair, incubation time per pair, and percentage of hatchling mortality compared to controls. Not all reproductive parameters were affected. Those traditionally regarded as indicators of reproductive capacity (number of eggs laid per clutch, number of eggs laid per pair, hatchlings per clutch, and fledglings per clutch) did not differ statistically between exposed and control birds. Findings support the hypothesis that very low PCB doses may be associated with endocrine disruption. It is suggested that evaluation of reproductive parameters related to parental care is more adequate to assess endocrine disruption than is evaluation of reproductive success parameters. Given its short breeding cycle, altricial breeding behavior, and other advantages not possessed by precocial birds, we propose using the zebra finch for evaluations of chemicals with endocrine-disruptive activity.

Cannabinoid inhibition improves memory in food-storing birds, but with a cost.

Food-storing birds demonstrate remarkable memory ability in recalling the locations of thousands of hidden food caches. Although this behaviour requires the hippocampus, its synaptic mechanisms are not understood. Here we show the effects of cannabinoid receptor (CB1-R) blockade on spatial memory in food-storing black-capped chickadees (Poecile atricapilla). Intra-hippocampal infusions of the CB1-R antagonist SR141716A enhanced long-term memory for the location of a hidden food reward, measured 72 h after encoding. However, when the reward location changed during the retention interval, birds that had received SR141716A during initial learning showed impairments in recalling the most recent reward location. Thus, blocking CB1-R activity may lead to more robust, long-lasting memories, but these memories may be a source of proactive interference. The relationship between trace strength and interference may be important in understanding neural mechanisms of hippocampal function in general, as well as understanding the enhanced memory of food-storing birds.

Where is the bird?

Patterns of song perception, learning, and expression differ across species, sexes, and individuals. We can understand the neurobiology of song better by paying attention to these differences. I focus selectively on a few of the studies done in my lab over in recent years to illustrate this.

Neural constraints on the complexity of avian song.

Why do birds sing? In many species, because the song attracts or retains a mate. Why do females pay attention? This paper reviews evidence that females may do so because male song can be an honest indicator of attributes of a male's brain that could contribute to his fitness or that of his young. Male songbirds learn and produce their songs using a set of brain regions collectively known as the song system. The learning has distinct auditory and motor components, and current data suggest that the neural changes that encode these forms of learning primarily occur in different subdivisions of the song system. There are positive correlations between song complexity and the volume of motor song system nucleus HVC, both between and within species. The correlations appear to arise because individual differences in volume lead to differences in capacity for learning. The differences in HVC volume are correlated with differences in the volumes of other song system components and with the volume of the forebrain. They are heritable. Thus, a complex song can be a signal to a female of immediate fitness (the male has a larger brain) and ultimate fitness (he has attractive characteristics that will be passed on to progeny).

Long-term memory for spatial locations in a food-storing bird (Poecile atricapilla) requires activation of NMDA receptors in the hippocampal formation during learning.

Food-storing birds use a form of long-term memory to recover their hidden food caches that depends on the hippocampal formation (HF). The authors assessed whether food-storing birds' long-term memory for spatial locations requires N-methyl-D-aspartate receptor (NMDA-R)-dependent synaptic plasticity. Black-capped chickadees (Poecile atricapilla) were given bilateral infusions of the NMDA-R antagonist AP5 into the hippocampus, and their memory on a spatial reference memory task was assessed. NMDA-R inactivation during learning prevented formation of long-term spatial memories but did not affect short-term memory and retrieval processes. NMDA-R inactivation immediately following learning did not disrupt long-term memory formation. NMDA-R inactivation disrupted the learning of multiple serially encoded reward locations when a 180-min delay separated successive learning episodes, suggesting that NMDA-R activity has a role in the incorporation of new information into existing long-term memory, as well as in forming unitary long-term memories.

Reversible inactivation of the hippocampal formation in food-storing black-capped chickadees (Poecile atricapillus).

The role of the hippocampal formation (HF) in memory processing was assessed in food-storing black-capped chickadees (Poecile atricapilla) by reversibly inactivating the HF during different memory tests. The memory tests required birds to remember a location based on spatial cues only, or based on a combination of both spatial and distinct visual cues. Inactivation of the HF impaired short-term spatial memory, but not visual-spatial memory. Inactivation of the HF impaired the retrieval of short-term (15 min) spatial memories, but not long-term (3-h) spatial memories. The pattern of deficits produced by inactivation of the HF in chickadees suggests a possible function of the hippocampal specialization of food-storing birds, as well as extends the notion of functional homology between the avian and mammalian HF.

Septum volume and food-storing behavior are related in parids.

The hippocampal formation (HF) of food-storing birds is larger than non-storing species, and the size of the HF in food-storing Black-Capped Chickadees (Poecile atricapillus) varies seasonally. We examined whether the volume of the septum, a medial forebrain structure that shares reciprocal connections with the HF, demonstrates the same species and seasonal variation as has been shown in the HF. We compared septum volume in three parid species; non-storing Blue Tits (Parus caeruleus) and Great Tits (Parus major), and food-storing Black-Capped Chickadees. We found the relative septum volume to be larger in chickadees than in the non-storing species. We also compared septum and nucleus of the diagonal band (NDB) volume of Black-Capped Chickadees at different times of the year. We found that the relative septum volume varies seasonally in food-storing birds. The volume of the NDB does not vary seasonally. Due to the observed species and seasonal variation, the septum, like the hippocampal formation of food-storing birds, may be specialized for some aspects of food-storing and spatial memory.

Song and the song control pathway in the brain can develop independently of exposure to song in the sedge warbler.

Previous studies have shown that female sedge warblers choose to mate with males that have more complex songs, and sexual selection has driven the evolution of both song complexity and the size of the major song control area (HVc) in the brain. In songbirds, learning from conspecifics plays a major role in song development and this study investigates the effects of isolation and exposure to song on song structure and the underlying song control system. Sibling pairs of hand-reared nestling sedge warblers were reared to sexual maturity under two conditions. Siblings in one group were reared individually in acoustic isolation in separate soundproof chambers. In the other group, siblings were reared together in an aviary with playback of recorded songs. The following spring, analysis of songs revealed that siblings reared in acoustic isolation produced normal song structures, including larger syllable repertoires than those exposed to song. We found no significant differences in the volumes of HVc, nucleus robustus archistnatalis, the lateral portion of the magnocellular nucleus and the density of dendritic spines between the two groups. Males exceeded females in all these measures, and also had a larger telencephalon. Our experiments show that complex song, sexual dimorphism in brain structure, and the size of song nuclei can all develop independently of exposure to song. These findings have important implications for how sexual selection can operate upon a complex male trait such as song and how it may also shape the more general evolution of brain structure in songbirds.