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.     

Recent experience modulates forebrain gene-expression in response to mate-choice cues in European starlings

Mate-choice decisions can be experience dependent, but we know little about how the brain processes stimuli that release such decisions. Female European starlings (Sturnus vulgaris) prefer males with long-bout songs over males with short-bout songs, and show higher expression of the immediate early gene (IEG) ZENK in the auditory forebrain when exposed to long-bout songs than when exposed to short-bout songs. We exposed female starlings to a short-day photoperiod for one of three durations and then, on an increased photophase, exposed them to one week of long-bout or short-bout song experience. We then examined their IEG response to novel long-bout versus novel short-bout songs by quantifying ZENK protein in two song-processing areas: the caudo-medial hyperstriatum ventrale and the caudo-medial neostriatum. ZENK expression in both areas increased with tenure on short-day photoperiods, suggesting that short days sensitize females to song. The ZENK response bias toward long-bout songs was greater in females with long-bout experience than in females with short-bout experience, indicating that the forebrain response bias toward a preferred trait depends on recent experience with that category of trait. This surprising level of neuroplasticity is immediately relevant to the natural history and fitness of the organism, and may underlie a mechanism for optimizing mate-choice criteria amidst locally variable distributions of secondary sexual characteristics.     

Response biases in auditory forebrain regions of female songbirds following exposure to sexually relevant variation in male song

In many species of songbirds, individual variation between the songs of competing males is correlated with female behavioral preferences. The neural mechanisms of song based female preference in songbirds are not known. Working with female European starlings (Sturnus vulgaris), we used immunocytochemistry for ZENK protein to localize forebrain regions that respond to sexually relevant variation in conspecific male song. The number of ZENK-ir cells in ventral caudo-medial neostriatum [NCMv] was significantly higher in females exposed to longer songs than in those exposed to shorter songs, whereas variation in the total duration of song exposure yielded no significant differences in ZENK expression. ZENK expression in caudo-medial ventral hyperstriatum [cmHV] was uniformly high in all subjects, and did not vary significantly among the three groups. These results suggest that subregions of NCM in female starlings are tuned to variation in male song length, or to song features correlated therewith. Female starlings exhibit robust behavioral preferences for longer over shorter male songs (Gentner and Hulse; Anim Behav 59:443-458, 2000). Therefore, the results of this study strongly implicate NCM in at least a portion of the perceptual processes underlying the complex natural behavior of female choice.     

Arrhythmic Song Exposure Increases ZENK Expression in Auditory Cortical Areas and Nucleus Taeniae of the Adult Zebra Finch

Rhythm is important in the production of motor sequences such as speech and song. Deficits in rhythm processing have been implicated in human disorders that affect speech and language processing, including stuttering, autism, and dyslexia. Songbirds provide a tractable model for studying the neural underpinnings of rhythm processing due to parallels with humans in neural structures and vocal learning patterns. In this study, adult zebra finches were exposed to naturally rhythmic conspecific song or arrhythmic song. Immunohistochemistry for the immediate early gene ZENK was used to detect neural activation in response to these two types of stimuli. ZENK was increased in response to arrhythmic song in the auditory association cortex homologs, caudomedial nidopallium (NCM) and caudomedial mesopallium (CMM), and the avian amygdala, nucleus taeniae (Tn). CMM also had greater ZENK labeling in females than males. The increased neural activity in NCM and CMM during perception of arrhythmic stimuli parallels increased activity in the human auditory cortex following exposure to unexpected, or perturbed, auditory stimuli. These auditory areas may be detecting errors in arrhythmic song when comparing it to a stored template of how conspecific song is expected to sound. CMM may also be important for females in evaluating songs of potential mates. In the context of other research in songbirds, we suggest that the increased activity in Tn may be related to the value of song for assessing mate choice and bonding or it may be related to perception of arrhythmic song as aversive.     

Complementary neural systems for the experience-dependent integration of mate-choice cues in European starlings

Choice of a particular mate phenotype may arise out of experience with the very phenotypes under consideration. Female European starlings (Sturnus vulgaris) prefer males that sing predominantly long-bout songs over males that sing predominantly short-bout songs, and thus, song-bout length is a phenotypic parameter instrumental in releasing the female's mate choice. The preferred long-bout songs induce higher expression of the immediate early gene (IEG) ZENK in the female auditory telencephalon than short-bout songs do, but this sensitivity to song length depends on the female's recent song experience. Here, we compared the experience-dependent modulation of ZENK with that of another IEG, FOS, and report that ZENK and FOS expression in the caudomedial mesopallium and caudomedial nidopallium show different modulation properties that complement natural variation in song-bout length. As reported previously, ZENK expression was greater in response to novel long-bout than to novel short-bout songs following a 1-week experience with long-bout but not short-bout songs. In contrast, FOS expression was greater in response to novel long-bout than to novel short-bout songs following a 1-week experience with short-bout but not long-bout songs. Thus, the ZENK and FOS signaling pathways are made sensitive to variation in song length by experiences with songs at opposite ends of the starling song-variation continuum, suggesting the presence of complementary neural systems made sensitive in register with the natural axis of phenotypic variation fundamental to the female's mate choice.     

Complementary neural systems for the experience‐dependent integration of mate‐choice cues in European starlings

Choice of a particular mate phenotype may arise out of experience with the very phenotypes under consideration. Female European starlings (Sturnus vulgaris) prefer males that sing predominantly long‐bout songs over males that sing predominantly short‐bout songs, and thus, song‐bout length is a phenotypic parameter instrumental in releasing the female's mate choice. The preferred long‐bout songs induce higher expression of the immediate early gene (IEG) ZENK in the female auditory telencephalon than short‐bout songs do, but this sensitivity to song length depends on the female's recent song experience. Here, we compared the experience‐dependent modulation of ZENK with that of another IEG, FOS, and report that ZENK and FOS expression in the caudomedial mesopallium and caudomedial nidopallium show different modulation properties that complement natural variation in song‐bout length. As reported previously, ZENK expression was greater in response to novel long‐bout than to novel short‐bout songs following a 1‐week experience with long‐bout but not short‐bout songs. In contrast, FOS expression was greater in response to novel long‐bout than to novel short‐bout songs following a 1‐week experience with short‐bout but not long‐bout songs. Thus, the ZENK and FOS signaling pathways are made sensitive to variation in song length by experiences with songs at opposite ends of the starling song‐variation continuum, suggesting the presence of complementary neural systems made sensitive in register with the natural axis of phenotypic variation fundamental to the female's mate choice. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2005     

Independent effects of song quality and experience with photostimulation on expression of the immediate,early gene ZENK (EGR‐1) in the auditory telencephalon of female European starlings

Age influences behavioral decisions such as reproductive timing and effort. In photoperiodic species, such age effects may be mediated, in part, by the individual's age‐accrued experience with photostimulation. In female European starlings (Sturnus vulgaris) that do not differ in age, experimental manipulation of photostimulation experience (photoexperience) affects hypothalamic, pituitary, and gonadal activity associated with reproductive development. Does photoexperience also affect activity in forebrain regions involved in processing a social cue, the song of males, which can influence mate choice and reproductive timing in females? Female starlings prefer long songs over short songs in a mate‐choice context, and, like that in other songbird species, their auditory telencephalon plays a major role in processing these signals. We manipulated the photoexperience of female starlings, photostimulated them, briefly exposed them to either long or short songs, and quantified the expression of the immediate‐early gene ZENK (EGR‐1) in the caudomedial nidopallium as a measure of activity in the auditory telencephalon. Using an information theoretic approach, we found higher ZENK immunoreactivity in females with prior photostimulation experience than in females experiencing photostimulation for the first time. We also found that long songs elicited greater ZENK immunoreactivity than short songs did. We did not find an effect of the interaction between photoexperience and song length, suggesting that photoexperience does not affect forebrain ZENK‐responsiveness to song quality. Thus, photoexperience affects activity in an area of the forebrain that processes social signals, an effect that we hypothesize mediates, in part, the effects of age on reproductive decisions in photoperiodic songbirds. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2009     

Stimulus-Evoked Increase of Glutamate in the Mediorostral Neostriatum/Hyperstriatum Ventrale of Domestic Chick After Auditory Filial Imprinting: An In Vivo Microdialysis Study

Abstract: Imprinting in chicks is a form of juvenile learning that has been used to study the basic cellular mechanisms of learning and memory. The forebrain area mediorostral neostriatum/hyperstriatum ventrale (MNH) is a center for acoustic imprinting. Electrophysiological and pharmacological behavioral studies in the MNH have demonstrated that the glutamatergic system and the associated receptors are critically involved in auditory filial imprinting. Accordingly, we investigated the hypothesis that stimulus-evoked glutamate release may be altered after this learning process. Using an in vivo microdialysis technique, we observed a significantly higher increase of extracellular glutamate level in tone-imprinted chicks during exposure to the previously imprinted tone than in socially imprinted control chicks. In a further series of experiments, where we exposed animals from both experimental groups to handling distress, glutamate levels in MNH showed only a slight increase, whereas we observed a pronounced increase of extracellular glutamate in the lobus parolfactorius (LPO), the avian analogue of the basal ganglia. No difference of distress-evoked glutamate release was found in MNH and LPO between tone-imprinted and socially imprinted chicks. The tone-evoked enhanced glutamate response in tone-imprinted chicks suggests that during auditory imprinting glutamatergic synapses develop the potential to increase transmitter release in response to the imprinting stimulus.     

Functional organization of the avian auditory field L

Summary The 2-deoxyglucose (2DG) autoradiographic method is used to analyse the functional organization of the auditory forebrain nucleus, field L, in parrots, ducks, pigeons, gulls and passerine birds. The data are compared to earlier studies in domestic and Guinea fowls. In all birds field L is a trilaminar structure, placed at the border between neostriatum mediale and caudale. The orientation and spatial extent within the forebrain, however, shows considerable variability. There is a close spatial relationship between field L and the overlying hyperstriatum ventrale, which is a secondary auditory center receiving input from field L. Stimulation with tones produces stripe like patterns of metabolic activity which are continuous across the layers of field L and the hyperstriatum ventrale. In all birds the position of the stripes in both areas shift in medio-lateral direction with decreasing tone frequency. In none of the birds the representation of frequencies above 3 kHz cover more than 20% of the neuronal space. Thus, high frequency hearing is underrepresented. Frequencies between 500 Hz and 3 kHz with somewhat variable representation, cover most of the neuronal space. Fowls and pigeons appear to have a low frequency specialization in field L.Abbreviations 2DG 2-deoxyglucose - FM frequency modulated     

Testosterone alters genomic responses to song and monoaminergic innervation of auditory areas in a seasonally breeding songbird

Behavioral responses to social stimuli often vary according to endocrine state. Our previous work has suggested that such changes in behavior may be due in part to hormone‐dependent sensory processing. In the auditory forebrain of female white‐throated sparrows, expression of the immediate early gene ZENK (egr‐1) is higher in response to conspecific song than to a control sound only when plasma estradiol reaches breeding‐typical levels. Estradiol also increases the number of detectable noradrenergic neurons in the locus coeruleus and the density of noradrenergic and serotonergic fibers innervating auditory areas. We hypothesize, therefore, that reproductive hormones alter auditory responses by acting on monoaminergic systems. This possibility has not been examined in males. Here, we treated non‐breeding male white‐throated sparrows with testosterone to mimic breeding‐typical levels and then exposed them to conspecific male song or frequency‐matched tones. We observed selective ZENK responses in the caudomedial nidopallium only in the testosterone‐treated males. Responses in another auditory area, the caudomedial mesopallium, were selective regardless of hormone treatment. Testosterone treatment reduced serotonergic fiber density in the auditory forebrain, thalamus, and midbrain, and although it increased the number of noradrenergic neurons detected in the locus coeruleus, it reduced noradrenergic fiber density in the auditory midbrain. Thus, whereas we previously reported that estradiol enhances monoaminergic innervation of the auditory pathway in females, we show here that testosterone decreases it in males. Mechanisms underlying testosterone‐dependent selectivity of the ZENK response may differ from estradiol‐dependent ones.© 2013 Wiley Periodicals, Inc. Develop Neurobiol 73: 455–468, 2013     

Development of song responses in the zebra finch caudomedial neostriatum: role of genomic and electrophysiological activities

Zebra finches first form demonstrable memories of specific songs between 25 and 35 days of age--several days after fledging from the nest. What accounts for the late onset of specific song memory formation? Here we investigated physiological development of the caudomedial neostriatum (NCM), part of the avian analogue of auditory cortex and a probable component of the system involved in song perception. Two types of physiological responses were characterized: electrophysiological (single-unit spike rate) and genomic (induction of the immediate early gene zenk, also known as zif-268, egr-1, ngfi-a, krox-24). We found that by day 20, zebra finches already have robust electrophysiological responses in NCM to song stimulation. Spike activity was greater in response to conspecific songs compared to heterospecific songs, white noise, or tones, and approximately 10% of the units showed selective responses to forward versus reversed songs. In contrast, at this age the zenk gene is expressed at a constitutively high level and undergoes no further induction in response to song presentation. At day 30, electrophysiological responses remained similar, but the zenk gene began to shift from a constitutive to an inducible pattern of expression. These results are consistent with a general role for NCM in the representation of song auditory patterns, and with a role for zenk gene expression in governing the efficiency of specific song memory storage at different ages.     

Neural response to bird’s own song and tutor song in the zebra finch field L and caudal mesopallium

Neurons in the song system nuclei of songbirds exhibit a strong preference for the sound of the birds own song relative to that of conspecific songs. This selectivity is observed in the high vocal center and the nucleus interface of the nidopallium, two song nuclei that receive input from the birds auditory system. To investigate the role of the auditory system in generating the selective responses observed in the song system, we recorded auditory responses in the zebra finch primary auditory forebrain, field L, and in a secondary auditory area, the caudal mesopallium. Field L and caudal mesopallium project directly or indirectly to the high vocal center and nucleus interface of the nidopallium and are presumed to provide substantial auditory input to the song system. We found that, on average, neurons in field L and caudal mesopallium did not show positive selective responses for the birds own song or tutor song relative to conspecific song. Moreover, there were no particular sub-areas in the auditory telencephalon that were relatively more selective than the average. The selectivity for the birds own song would therefore be restricted to song nuclei and would arise in one processing step, potentially found at the interface between the auditory and the song systems.Abbreviations BOS birds own song - CM caudal mesopallium (older term: caudal hyperstriatum ventrale or cHV) - Con conspecific song - HVC high vocal center - LMAN lateral magnocellular nucleus of the anterior nidopallium (older term: lateral magnocellular nucleus of the anterior neostriatum) - LPS pallial-subpallial lamina (older term: lamina medularis dorsalis or LMD) - NCM caudal medial nidopallium (older term: caudo-medial neostriatum) - NIf nucleus interface of the nidopallium (older term: nucleus interface of the neostriatum) - RA robust nucleus of arcopallium (older term: robust nucleus of the archistriatum) - Rev reverse BOS - Revorder reverse order of BOS - Uva nucleus uvaeformis of the thalamusNew avian brain terminology has been used in this paper (). Older terms are given in parentheses in the list of abbreviations     

Gap detection in the starling (Sturnus vulgaris)

Summary Gap-detection thresholds of single units were determined from auditory forebrain neurons of the awake starling. Nine different response types were statistically defined from the discharge pattern to a 400 ms broadband noise stimulus. The gap stimuli consisted of two broadband noise bursts which were separated by a gap ranging from 0.4 to 204.8 ms duration. The median minimumdetectable gap for 121 out of 145 units that had a significant threshold 204.8ms was 12.8 ms; 20% of the neurons showed thresholds between 0.4 and 3.2 ms. The neurons of the nine response types differed significantly in their minimum-detectable gaps; neurons with phasic-tonic and phasic excitation exhibited the best (i.e. shortest) minimum-detectable gaps. The neurons of the three different recording areas (field L, NCM and HV) were significantly different in their minimumdetectable gaps; field L neurons showed the best temporal resolution for gaps in broadband noise. Gap-detection thresholds are compared with psychophysical thresholds determined with the same stimuli and the relevance of forebrain units for temporal resolution is discussed.Abbreviations CS control stimulus - HV hyperstriatum ventrale - HVc hyperstriatum ventrale pars caudalis - NB noise burst - NCM neostriatum caudale pars medialis - NS noise stimulus - SGS standard gap series - TW time window     

The hippocampus and caudomedial neostriatum show selective responsiveness to conspecific song in the female zebra finch

The perception of song is vital to the reproductive success of both male and female songbirds. Several neural structures underlying this perception have been identified by examining expression of immediate early genes (IEGs) following the presentation of conspecific or heterospecific song. In the few avian species investigated, areas outside of the circuit for song production contain neurons that are active following song presentation, specifically the caudal hyperstriatum ventrale (cHV) and caudomedial neostriatum (NCM). While studied in detail in the male zebra finch, IEG responses in these neural substrates involved in song perception have not been quantified in females. Therefore, adult female zebra finches were presented with zebra finch song, nonzebra finch song, randomly generated tones, or silence for 30 min. One hour later they were sacrificed, and their brains removed, sectioned, and immunocytochemically processed for FOS expression. Animals exposed to zebra finch song had a significantly higher density of FOS-immunoreactive cells in the NCM than those presented with other songs, tones, or silence. Neuronal activation in the cHV was equivalent in birds that heard zebra finch and non-zebra finch song, expression that was higher than that observed in the groups that heard no song. Interestingly, the hippocampus (HP) and adjacent parahippocampal area (AHP) were activated in a manner comparable to the NCM. These results suggest a general role for the cHV in song perception and a more specific role for the NCM and HP/AHP in facilitating recognition of and responsiveness to species-specific song in female zebra finches.     

Calbindin-positive neurons reveal a sexual dimorphism within the songbird analogue of the mammalian auditory cortex

The oscine song system, a set of interconnected brain nuclei involved in song production and learning, is one of the first and clearest examples of brain sexual dimorphism in a vertebrate, being typically well-developed in males, but not females. Here we present evidence for a sexual dimorphism in the caudomedial nidopallidum (NCM), an auditory area outside of the song system. NCM is thought to correspond to a portion of the auditory cortex of mammals and is involved in the perceptual processing of birdsong. We show that cells immunolabeled for the calcium-binding protein calbindin are primarily localized to caudal NCM and are almost twice as numerous in males as in females. We demonstrate that calbindin-positive cells constitute a subset of GABAergic cells in NCM, and show that the sex dimorphism in this cell population does not result from local gender differences in the overall density of neuronal or GABAergic cells. In addition, we demonstrate that calbindin-positive cells lack song-induced expression of the activity-dependent gene ZENK, and that song stimulation does not change the density or distribution of these cells in NCM. Finally, we show that the distribution of calbindin-positive cells in NCM is strikingly similar to the mRNA expression for the estrogen-generating enzyme aromatase. Together these results suggest that NCM is likely composed of neurochemically-distinct domains and presents a marked sex dimorphism in a specific subset of GABAergic neurons, which may confer sex-specific sensory processing capabilities to this auditory area. Our results also suggest that local sex steroid hormones may play a local role in auditory processing in the songbird telencephalon.     

Calbindin‐positive neurons reveal a sexual dimorphism within the songbird analogue of the mammalian auditory cortex

The oscine song system, a set of interconnected brain nuclei involved in song production and learning, is one of the first and clearest examples of brain sexual dimorphism in a vertebrate, being typically well‐developed in males, but not females. Here we present evidence for a sexual dimorphism in the caudomedial nidopallidum (NCM), an auditory area outside of the song system. NCM is thought to correspond to a portion of the auditory cortex of mammals and is involved in the perceptual processing of birdsong. We show that cells immunolabeled for the calcium‐binding protein calbindin are primarily localized to caudal NCM and are almost twice as numerous in males as in females. We demonstrate that calbindin‐positive cells constitute a subset of GABAergic cells in NCM, and show that the sex dimorphism in this cell population does not result from local gender differences in the overall density of neuronal or GABAergic cells. In addition, we demonstrate that calbindin‐positive cells lack song‐induced expression of the activity‐dependent gene ZENK, and that song stimulation does not change the density or distribution of these cells in NCM. Finally, we show that the distribution of calbindin‐positive cells in NCM is strikingly similar to the mRNA expression for the estrogen‐generating enzyme aromatase. Together these results suggest that NCM is likely composed of neurochemically‐distinct domains and presents a marked sex dimorphism in a specific subset of GABAergic neurons, which may confer sex‐specific sensory processing capabilities to this auditory area. Our results also suggest that local sex steroid hormones may play a local role in auditory processing in the songbird telencephalon. © 2005 Wiley Periodicals, Inc. J Neurobiol, 2005     

Social context-induced song variation affects female behavior and gene expression

Social cues modulate the performance of communicative behaviors in a range of species, including humans, and such changes can make the communication signal more salient. In songbirds, males use song to attract females, and song organization can differ depending on the audience to which a male sings. For example, male zebra finches (Taeniopygia guttata) change their songs in subtle ways when singing to a female (directed song) compared with when they sing in isolation (undirected song), and some of these changes depend on altered neural activity from a specialized forebrain-basal ganglia circuit, the anterior forebrain pathway (AFP). In particular, variable activity in the AFP during undirected song is thought to actively enable syllable variability, whereas the lower and less-variable AFP firing during directed singing is associated with more stereotyped song. Consequently, directed song has been suggested to reflect a “performance” state, and undirected song a form of vocal motor “exploration.” However, this hypothesis predicts that directed–undirected song differences, despite their subtlety, should matter to female zebra finches, which is a question that has not been investigated. We tested female preferences for this natural variation in song in a behavioral approach assay, and we found that both mated and socially naive females could discriminate between directed and undirected song—and strongly preferred directed song. These preferences, which appeared to reflect attention especially to aspects of song variability controlled by the AFP, were enhanced by experience, as they were strongest for mated females responding to their mate's directed songs. We then measured neural activity using expression of the immediate early gene product ZENK, and found that social context and song familiarity differentially modulated the number of ZENK-expressing cells in telencephalic auditory areas. Specifically, the number of ZENK-expressing cells in the caudomedial mesopallium (CMM) was most affected by whether a song was directed or undirected, whereas the caudomedial nidopallium (NCM) was most affected by whether a song was familiar or unfamiliar. Together these data demonstrate that females detect and prefer the features of directed song and suggest that high-level auditory areas including the CMM are involved in this social perception.     

Social Context–Induced Song Variation Affects Female Behavior and Gene Expression

Social cues modulate the performance of communicative behaviors in a range of species, including humans, and such changes can make the communication signal more salient. In songbirds, males use song to attract females, and song organization can differ depending on the audience to which a male sings. For example, male zebra finches (Taeniopygia guttata) change their songs in subtle ways when singing to a female (directed song) compared with when they sing in isolation (undirected song), and some of these changes depend on altered neural activity from a specialized forebrain-basal ganglia circuit, the anterior forebrain pathway (AFP). In particular, variable activity in the AFP during undirected song is thought to actively enable syllable variability, whereas the lower and less-variable AFP firing during directed singing is associated with more stereotyped song. Consequently, directed song has been suggested to reflect a “performance” state, and undirected song a form of vocal motor “exploration.” However, this hypothesis predicts that directed–undirected song differences, despite their subtlety, should matter to female zebra finches, which is a question that has not been investigated. We tested female preferences for this natural variation in song in a behavioral approach assay, and we found that both mated and socially naive females could discriminate between directed and undirected song—and strongly preferred directed song. These preferences, which appeared to reflect attention especially to aspects of song variability controlled by the AFP, were enhanced by experience, as they were strongest for mated females responding to their mate's directed songs. We then measured neural activity using expression of the immediate early gene product ZENK, and found that social context and song familiarity differentially modulated the number of ZENK-expressing cells in telencephalic auditory areas. Specifically, the number of ZENK-expressing cells in the caudomedial mesopallium (CMM) was most affected by whether a song was directed or undirected, whereas the caudomedial nidopallium (NCM) was most affected by whether a song was familiar or unfamiliar. Together these data demonstrate that females detect and prefer the features of directed song and suggest that high-level auditory areas including the CMM are involved in this social perception.     

Song Competition Affects Monoamine Levels in Sensory and Motor Forebrain Regions of Male Lincoln's Sparrows (Melospiza lincolnii)

Male animals often change their behavior in response to the level of competition for mates. Male Lincoln''s sparrows (Melospiza lincolnii) modulate their competitive singing over the period of a week as a function of the level of challenge associated with competitors'' songs. Differences in song challenge and associated shifts in competitive state should be accompanied by neural changes, potentially in regions that regulate perception and song production. The monoamines mediate neural plasticity in response to environmental cues to achieve shifts in behavioral state. Therefore, using high pressure liquid chromatography with electrochemical detection, we compared levels of monoamines and their metabolites from male Lincoln''s sparrows exposed to songs categorized as more or less challenging. We compared levels of norepinephrine and its principal metabolite in two perceptual regions of the auditory telencephalon, the caudomedial nidopallium and the caudomedial mesopallium (CMM), because this chemical is implicated in modulating auditory sensitivity to song. We also measured the levels of dopamine and its principal metabolite in two song control nuclei, area X and the robust nucleus of the arcopallium (RA), because dopamine is implicated in regulating song output. We measured the levels of serotonin and its principal metabolite in all four brain regions because this monoamine is implicated in perception and behavioral output and is found throughout the avian forebrain. After controlling for recent singing, we found that males exposed to more challenging song had higher levels of norepinephrine metabolite in the CMM and lower levels of serotonin in the RA. Collectively, these findings are consistent with norepinephrine in perceptual brain regions and serotonin in song control regions contributing to neuroplasticity that underlies socially-induced changes in behavioral state.     

The integration of song environment by catecholaminergic systems innervating the auditory telencephalon of adult female European starlings

Mate choice is among the most consequential decisions a sexually reproducing organism can make. In many songbird species, females make mate-choice decisions based, in part, on variation between males in songs that reflect their quality. Importantly, females may adjust their choice relative to the prevalence of high quality songs. In European starlings (Sturnus vulgaris), females prefer males that primarily sing long songs over those that primarily sing short songs, and sensitivity of the auditory telencephalon to song length depends on the prevalence of long songs in the environment. Several lines of evidence suggest a role for noradrenergic innervation of the auditory telencephalon in mediating this neuro- and behavioral plasticity. To simulate variation in quality of the song environment, we exposed adult female starlings to 1 week of either long or short songs and then quantified several monoamines and their metabolites in the caudomedial mesopallium and caudomedial nidopallium (NCM) using high performance liquid chromatography. We also used immunocytochemistry to assess these areas for immunoreactive dopamine-beta-hydroxylase (DBH-ir), the enzyme that synthesizes norepinephrine. We found that long songs elevated levels of the principal norepinephrine metabolite, the principal dopamine metabolite, and the probability of DBH-ir in the NCM compared to short songs. Song environment did not appear to influence norepinephrine or dopamine levels. Thus, the quality of the song environment regulates the local secretion of catecholamines, particularly norepinephrine, in the female auditory telencephalon. This may form a basis for plasticity in forebrain sensitivity and mate-choice behavior based on the prevalence of high-quality males.