Song- and order-selective neurons develop in the songbird anterior forebrain during vocal learning

The anterior forebrain (AF) pathway of songbirds has an essential but poorly understood function during song learning, a process requiring auditory experience. Consistent with a role in processing auditory information, two nuclei of the AF, the lateral magnocellular nucleus of the anterior neostriatum (IMAN) and Area X (X), contain some of the most complex auditory neurons known. In adult zebra finches, these neurons are strongly selective for both spectral and temporal properties of song: They respond more robustly to the bird's own song (BOS) than to songs of conspecific individuals, and they respond less well to BOS if it is played in reverse. IMAN and X neurons of young finches early in the process of song learning (30-45 days of age) are also song responsive, but lack the song and order selectivity present in adult birds. By an intermediate stage of learning (60 days), when birds have experience of both tutor song and their own developing (plastic) song, AF neurons have significant song and order selectivity for both tutor song and BOS (in this case, plastic song). The degree of BOS selectivity is still less than that found in adults, however. In addition, neurons at 60 days are heterogenous in their preference for BOS versus tutor song: Most prefer BOS, some prefer tutor song, and others respond equally to both songs. The selectivity of adult AF auditory neurons therefore arises rapidly during development in neurons that are initially unselective. These neurons are one of the clearest examples of experience-dependent acquisition of complex stimulus selectivity. Moreover, the neural selectivity for both BOS and tutor song at 60 days raises the possibility that experience of both songs during learning contributes to the properties of individual AF neurons.     

Central DSP-4 treatment decreases norepinephrine levels and courtship behavior in male zebra finches

In zebra finches, gonadal steroids activate male courtship, including singing, and also strongly modulate norepinephrine (NE) levels and turnover in brain areas regulating courtship behavior. In a previous study, systemic administration of DSP-4 caused significant decreases in courtship singing. These behavioral decrements were correlated with the degree of NE depletion in several vocal control nuclei. In the present study, we attempted to further decrease brain NE levels while minimizing systemic effects by infusing DSP-4 directly into the third ventricle. DSP-4 treatment significantly reduced NE levels in three of six vocal control nuclei and both hypothalamic nuclei sampled without significantly altering dopamine or serotonin levels in any areas. DSP-4-treated males took longer to begin singing and performed fewer song bouts and courtship displays. Interestingly, behavioral deficits were limited to courtship song displays, other behavior patterns, including female-directed behaviors like approach and follow, were unaffected by DSP-4 treatment. DSP-4 treatment appeared to affect singing behavior by causing deficits in initial attentiveness to females and initiation of singing rather than by affecting song structure.     

Distribution of GABA-like immunoreactivity in the song system of the zebra finch

GABA-like immunoreactivity (GABA-LIR) was mapped in the male and female zebra finch song system using a polyclonal antibody to GABA. GABA-LIR was found throughout the song system in neurons and neuropil of the robust nucleus of the archistriatum (RA), the higher vocal center (HVC), Area X, the magnocellular nucleus of the neostriatum (MAN), and the dorsomedial portion of the nucleus intercollicularis (DM of ICo). Puncta present in the lateral division of MAN (lMAN) may be local interneurons since the only known afferents of lMAN are from the dorsolateral nucleus of the anterior thalamus (DLM), which did not appear to have any cell bodies with GABA-LIR. Distinct and dense puncta with GABA-LIR were present in DLM, and may be projections from Area X/lobus parolfactorius (LPO). Dramatic sex differences in GABA-LIR distribution were found. Females did not appear to have any GABA-LIR above background in either RA or HVC. Females also did not appear to have a distinct Area X, although they did have many small, lightly staining cell bodies in the corresponding LPO. The distribution of GABA-LIR and sex differences in its distribution suggests that GABAergic neurons may play a role in the acquisition and/or production of song in the zebra finch.     

Melatonin binding in the house sparrow song control system: sexual dimorphism and the effect of photoperiod

Avian song is a sexually dimorphic behavior which is regulated seasonally. This regulation involves the construction and growth of song control structures: the high vocal center (HVC), nucleus robustus archistrialis (RA), nucleus magnocellularis anterior (MAN), and Area X. Song behavior and its neural correlates are controlled by steroid-dependent and independent processes. The avian circadian system is known to be involved in both daily processes and seasonal reproduction. A major part of this system is the circadian secretion of melatonin by the pineal gland. To determine possible interactions of the circadian and song control systems, the distribution and density of 2-[125I]iodomelatonin (IMEL) binding, an indicator of melatonin sensitivity, were determined in male and female house sparrow brains. Specific binding was found in visual system centers of both genders, but binding in HVC, RA, and Area X was present only in males. Binding in MAN was present in both sexes. Although the effects of short and long photoperiods on male house sparrow IMEL binding in song structures revealed no systematic changes, there were significant differences in binding under different photoperiods in HVC and RA. IMEL binding in the tectofugal nucleus rotundus, however, was consistently highest under short day conditions. IMEL binding in song control nuclei was independent of testicular influence, since castration did not affect it significantly. The data point to a role for the circadian system of house sparrows in song control, but a specific role for melatonin in the daily or seasonal regulation of the song control system in birds, could not be determined.     

Axonal connections of the high vocal center and surrounding cortical regions in juvenile and adult male zebra finches

Neuronal connections of the High Vocal Center (HVC), a cortical nucleus of songbirds necessary for learned vocal behavior, and the region adjacent to HVC called paraHVC (pHVC), were studied in adult and juvenile male zebra finches. Extremely small injections of fluorescent dextran amines or biocytin were made within subregions of HVC and pHVC to define the precise nature and development of these pathways. In adults, all HVC injections produced an even, nontopographic distribution of retrograde label throughout the medial magnocellular nucleus of the anterior neostriatum (mMAN), the interfacial nucleus (NIf), and the uvaeform nucleus of the thalamus (Uva) and an even distribution of anterograde label within area X of the striatum and the robust nucleus of the archistriatum (RA). These same patterns of projections were present in juvenile birds 20-23 days of age, including the projection from HVC to RA, which has previously been reported to develop only after 25-30 days of age. Results also establish a novel efferent projection from HVC to pHVC in both juvenile and adult birds. Injections into pHVC indicate that this region receives afferent input from song control areas HVC, mMAN, medial regions of the parvicellular shell of lateral MAN, NIf, and Uva and projects to Area X, caudomedial regions of striatum, and regions of the caudomedial neostriatum (NCM). Thus, neuronal connections of pHVC are highly integrated with circuitry important for vocal behavior and are distinct from those of HVC. Such differences establish HVC and pHVC as separate brain areas and suggest that each may serve a different function in vocal behavior. Control injections in both juveniles and adults produced specific patterns of projections from areas outside of HVC to areas outside of RA, illustrating an overall spatial organization of projections from HVC and neighboring cortical areas. Further, although neuronal connections of HVC are not topographic, projections of HVC, pHVC, and surrounding areas demonstrate a broad spatial organization of efferents to striatum and regions surrounding RA, thus defining a level of organization beyond that of individual song control nuclei.     

An improved method for the simultaneous analysis of norepinephrine, epinephrine and dopamine at the picogram level by mass fragmentography (author's transl)

Gas chromatography-mass spectrometry makes possible the simultaneous measurement of norepinephrine (NE), epinephrine (E) and dopamine (DA) at the picogram level. The equipment used was the Shimadzu LKB 9000 GC-MS (MID-MD. The TFA derivatives of the compounds were analysed by the GC-MS system equipped with a 3ft X 3mm column packed with 1.5%OV-1 and the temperature was programed from 135 degrees to 260 degrees C at 10 degrees C/min increments. The mass spectrum showed molecular ions at m/e 552, 566 and 440 which corresponds to the TFA derivatives of NE, E and DA respectively. The base peaks of NE, E and DA were m/e 109, 140 and 328 respectively. The peaks at m/e 126 (NE and DA) and 140 (E) were applied to avail themselves of simultaneous multiple-ion analyses by mass fragmentography. As little as 4pg of the compounds injected into the column could be measured with good reproducibility, and the linearity of the response was maintained up to 120 pg. The sensitivity was of the order of ng or pg which enable quantitation with 1ml of human serum and urine sample. The procedures were simpler and less time consuming than previously reported methods. Recovery rates of NE, E and DA from serum were 25.7%, 13.7% and 76.1%, whereas those from urine were 100%, 89.6% and 100% respectively. The specificity of this method surpasses and cannot be compared to any other existing quantitative methods.     

Management of the patient with sickle cell disease

A discrete neural circuit mediates the production of learned vocalizations in oscine songbirds. Although this circuit includes some bilateral pathways at midbrain and medullary levels, the forebrain components of the song control network are not directly connected across the midline. There have been no previous reports of bilateral projections from medullary and midbrain vocal control nuclei back to the forebrain song system, but the existence of such bilateral corollary discharge pathways was strongly suggested by the recent observation that unilateral stimulation of a forebrain song nucleus during singing leads to a rapid readjustment of premotor activity in the contralateral forebrain. In the present study, we used neuroanatomical tracers to demonstrate bilateral projections from (a) the rostral ventrolateral medulla (RVL), which may control respiratory aspects of vocalization, to nucleus uvaeformis (Uva), and (b) the dorsomedial intercollicular nucleus (DM), a midbrain vocal control region, to Uva. Both RVL and DM receive descending projections from the forebrain song nucleus robustus archistriatalis, and Uva projects directly to the forebrain song nuclei interfacialis and high vocal center. We suggest that the bilateral feedback projections from DM and RVL to Uva function to coordinate the two hemispheres during singing in adult songbirds and to convey internal feedback of premotor signals to the forebrain in young birds that are learning to sing.     

Noradrenergic system of the zebra finch brain: immunocytochemical study of dopamine-beta-hydroxylase

Oscine birds are among the few animal groups that have vocal learning, and their brains contain a specialized system for song learning and production. We describe here the immunocytochemical distribution of dopamine-beta-hydroxylase (DBH), a noradrenergic marker, in the brain of an oscine, the zebra finch (Taeniopygia guttata). DBH-positive cells were seen in the locus coeruleus, the nucleus subcoeruleus ventralis, the nucleus of the solitary tract, and the caudolateral medulla. Immunoreactive fibers and varicosities had a much wider brain distribution. They were particularly abundant in the hippocampus, septum, hypothalamus, area ventralis of Tsai, and substantia nigra, where they formed dense pericellular arrangements. Significant immunoreactivity was observed in auditory nuclei, including the nucleus mesencephalicus lateralis pars dorsalis, the thalamic nucleus ovoidalis, field L, the shelf of the high vocal center (HVC), and the cup of the nucleus robustus archistriatalis (RA), as well as in song control nuclei, including the HVC, RA, the lateral magnocellular nucleus of the anterior neostriatum, and the dorsomedial nucleus (DM) of the intercollicular complex. Except for the DM, DBH immunoreactivity within song nuclei was comparable to that of surrounding tissues. Conspicuously negative were the lobus paraolfactorius, including song nucleus area X, and the paleostriatum. Our results are in agreement with previous studies of the noradrenergic system performed in nonoscines. More importantly, they provide direct evidence for a noradrenergic innervation of auditory and song control nuclei involved in song perception and production, supporting the notion that noradrenaline is involved in vocal communication and learning in oscines.     

Age, side height, and spindle shape of the crib in climbing over the side

Norepinephrine (NE) was microinjected into the paraventricular nucleus (PVN), while microdialysis was used to monitor extracellular dopamine (DA) and acetylcholine (ACh) in the nucleus accumbens (NAc). The PVN is a site where exogenously administered NE can act through alpha 2 receptors to elicit eating behavior and preference for carbohydrates. It was hypothesized that NE in the PVN acts on a behavior reinforcement system by altering the DA/ACh balance in the NAc. NE microinjections (80 nmol in 0.3 microliter), which effectively elicited feeding in satiated rats in a separate test, caused a significant increase in extracellular DA (109%) and decrease in ACh (-27%) when the same animals were tested in the absence of food. In contrast when the food was available and ingested, ACh increased (51%) instead of decreasing. These results support the hypothesis that a functional link exists between the PVN and the NAc in which DA helps initiate and ACh helps stop appetitive behavior involved in the reinforcement of eating.     

Evidence for Opioid Involvement in the Regulation of Song Production in Male European Starlings (Sturnus vulgaris).

Many social animals vocalize at high rates, suggesting that vocal communication is highly motivated and rewarding. In songbirds, much is known about the neural control of vocal behavior; however, little is known about neurobiological mechanisms regulating the motivation to communicate. This study examined a possible role for opioid neuropeptides in motivation and reward associated with song production in male European starlings (Sturnus vulgaris). Peripheral opioid blockade facilitated male song production. Furthermore, methionine-enkephalin immunolabeled fiber densities within brain regions in which opioids are known to regulate motivation and reward (i.e., the medial preoptic nucleus and ventral tegmental area) related positively to male song production. These data suggest that song production might be regulated by opioid activity within motivation and reward neural systems. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Lesions of the substantia nigra retard Pavlovian eye-blink but not heart rate conditioning in the rabbit.

Bilateral radio-frequency (RF) lesions of the substantia nigra retarded Pavlovian eye-blink (EB) conditioning without affecting concurrent heart rate (HR) conditioning. Dopamine (DA) depletion occurred only in the caudate nucleus, whereas norepinephrine (NE) depletion was limited to the hypothalamus. Bilateral 6-hydroxydopamine (6-OHDA) lesions of substantia nigra retarded acquisition of both EB and HR responses. Six-OHDA lesions produced significant NE depletion in the nucleus acumbens/septi, frontal cortices, and hypothalamus, as well as DA depletion in the caudate nucleus. Trials required to reach EB conditioning criterion were significantly correlated to the caudate DA levels. The magnitude of conditioned bradycardia was on the other hand significantly correlated with hypothalamic NE levels. Results suggest that interruption of the nigro-striatal dopaminergic pathway retards Pavlovian somatomotor learning without affecting concurrent autonomic learning, although the latter may depend on an intact ascending NE pathway to the hypothalamus, which passes through the tegmentum and thus is also destroyed in some cases by substantia nigra lesions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Sex difference in the size of the neural song control regions in a dueting songbird with similar song repertoire size of males and females

Previous studies have suggested a causal relation between sex differences in behavior such as singing and sex differences in the size of brain areas such as the forebrain song control areas of songbirds. In the present study we show that the size of the forebrain vocal control areas nucleus hyperstriatalis ventrale pars caudale (HVC) and nucleus robustus archistriatalis (RA) and its neuron numbers are about twice as large in males as in females of the African dueting bush shrike Laniarius funebris. However, song types are of similar complexity (number of elements per song type, physical properties of elements) in both sexes, and repertoire size does not differ between males and females. Furthermore, in captivity male and female shrikes are able to learn the same song types. This demonstrates for the shrike that sex differences in the size of vocal control areas and in its neuron numbers do not predict the type of sex-typical vocal behavior. This result is supported by a statistical comparison of the sex differences in HVC size, RA size, and song repertoire size of all songbird species studied to date. Sex differences in species in which only the males sing are indeed larger than in species in which the females also sing; in songbird species with singing females, however, the sex differences in HVC and RA volume appear to be independent of the vocal repertoire size of females. The songbird model therefore does not support the notion that sex differences in area size and neuron number explain sex differences in a behavior that occurs in both sexes. Furthermore, in the shrike, neuron soma size is similar in males and females in the song motonucleus hypoglossus pars tracheosyringealis (nXIIts) and in the premotor nucleus RA, but is sexually dimorphic in the higher vocal center HVC. Thus, male and female shrikes produce songs of similar complexity with different neuron phenotypes.     

Behavioral and neurochemical alterations following intracerebroventricular administration of anti-serotonin antibodies in adult Balb/c mice

The effects of intracerebroventricular injections of serotonin (5-HT) antibodies were studied for changes in 5-HT, dopamine (DA), their metabolites and norepinephrine (NE) as well as 5-HT mediated behavior in adult mice. While nociceptive thresholds (tail-flick latency) were inhibited in antibody treated animals, tremor response to 5-methoxy-N,N-dimethyl tryptamine administration was increased. 5-HT and DA in the nucleus raphe dorsalis (NRD), substantia nigra (SN), nucleus caudatus putamen (NCP) and in the substantia grisea centralis, and NE in the former two nuclei were significantly decreased in these animals. 5-Hydroxyindoleacetic acid was unaffected in all nuclei except NRD, where it was inhibited. Homovanillic acid and 3,4-dihydroxyphenylacetic acid were inhibited in all nuclei except in NCP. The brunt of insult was more evident in NRD and SN where all neurotransmitters were inhibited for a longer period. 5-HT turnover was increased in all the nuclei, however only SN showed increased DA turnover. It may be assumed that the observed neurochemical and behavioral changes were the consequence of the antibodies binding to 5-HT, which in turn influenced the anatomically and functionally connected neurotransmitters. While the study contributes to the existing understanding of central neurotransmitter control on behavior, it fails to delineate the underlying mechanism. The possibility of developing a useful, drug-free 5-HT deficient animal model for studying clinical disorders, as well as for solving some of the basic questions related to the physiological functions of 5-HT in adult animals are envisaged from the study.     

Lesions to the medial preoptic nucleus differentially affect singing and nest box-directed behaviors within and outside of the breeding season in European starlings (Sturnus vulgaris).

Little is known about how the brain regulates context-appropriate communication. European starlings produce song in various social contexts. During the breeding season, males with nest sites sing high levels of sexually motivated song in response to a female. Outside of this context, song rates are not affected by female presence. The medial preoptic nucleus (POM) regulates male sexual behavior, and studies in songbirds implicate the POM in sexually motivated song. Recent data suggest that the role of the POM might extend to song produced in other contexts as well. To examine this possibility, effects of bilateral electrolytic lesions of the POM on singing and other behaviors in adult male starlings within sexually relevant and nonsexual contexts were studied. Lesions to the POM exclusively reduced song and nest box-directed behaviors within highly sexually relevant contexts. Unexpectedly, POM lesions increased song in a nonsexual context, suggesting an inhibitory role for the POM in this context. These data suggest that the POM interacts with the song control system so that song occurs in an appropriate social context in response to appropriate stimuli. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Photoperiod and testosterone independently affect vocal control region volumes in adolescent male songbirds

Previously, we found that, unlike adults, adolescent male dark-eyed juncos (Junco hyemalis) maintained large Area X volumes despite having low plasma testosterone concentrations. Other studies indicate that photoperiod may act independently of testosterone to modulate vocal control region (VCR) volumes in adult songbirds. In the present study, we investigated the effects of testosterone and photoperiod on the volumes of four VCRs in adolescent male juncos. To test the hypothesis that VCR volumes in these males are testosterone independent, we treated birds exposed to short days with testosterone and later compared their VCR volumes with those of birds exposed to short days without testosterone. To examine whether photoperiod alone could affect VCR volumes independent of testosterone, we measured these volumes in photorefractory birds exposed to long photoperiod without testosterone. Administering testosterone induced singing, yet increased the volume of only one VCR, the robust nucleus of the anterior neostriatum (RA). In contrast, long photoperiod increased several VCR volumes (Area X, higher vocal center, and RA) despite low testosterone levels, but did not induce singing. Our results suggest a limited role for testosterone, but an important role for photoperiod, in controlling VCR volumes in adolescent male juncos. In addition, the results demonstrate that singing behavior can be induced in adolescent males without a concomitant increase in most VCR volumes.     

Central dopamine turnover in guinea pig pups during separation from their mothers in a novel environment.

Guinea pig pups that were separated from their mothers and placed into a novel environment for 90 min showed an increase in dopamine (DA) turnover (ratio of metabolites to DA) in the septum compared with undisturbed baseline controls. Pups placed into the novel environment with their mothers exhibited an intermediate level of DA turnover. After 24 hr of separation in the novel environment, pups' DA turnover in the septum had returned to the baseline level. DA turnover in the caudate nucleus was unaffected by these procedures. Also, turnover in both septum and caudate nucleus when pups were not separated was positively correlated with the number of vocalizations emitted during 30 min of separation. These results closely parallel findings in separated monkeys and indicate that the guinea pig represents a useful rodent model for studying such effects. That elevated DA turnover during separation occurred in the septum suggests involvement of the mesolimbic system. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Soldiers, surgeons and the campaigns to combat sexually transmitted diseases in colonial India, 1805-1860

Here, we examine the connectivity of two previously identified telencephalic stations of the auditory system of adult zebra finches, the neostriatal "shelf" that underlies the high vocal center (HVC) and the archistriatal "cup" adjacent to the robust nucleus of the archistriatum (RA). We used different kinds of neuroanatomical tracers to visualize the projections from the shelf to the HVC. In addition, we show that the shelf projects to the cup and that the cup projects to thalamic, midbrain, and pontine nuclei of the ascending auditory pathway. Our observations extend to songbirds anatomical features that are found in the auditory pathways of a nonoscine bird, the pigeon (Wild et al. [1993] J. Comp. Neurol. 337:32-62), and we suggest that the descending auditory projections found in mammals may also be a general property of the avian brain. Finally, we show that the oscine song control system is closely apposed to auditory pathways at many levels. Our observations may help in understanding the evolution and organization of networks for vocal communication and vocal learning in songbirds.     

Catecholamine depletion in mice upon reexposure to stress: Mediation of the escape deficits produced by inescapable shock.

Reports 9 experiments with 372 male Swiss-Webster mice in which, immediately following exposure to 60 inescapable shocks, Ss had significantly reduced hypothalamic norepinephrine (NE). Within 24 hrs NE levels returned to control values. Reexposure to as few as 10 shocks 24 hrs after initial stress exposure resulted in significant decline of NE. At this interval after shock, escape performance was severely disrupted, with a large proportion of Ss exhibiting numerous failures to escape shock. Increasing brain dopamine (DA) and NE by levodopa treatment prior to shock prevented the escape deficits. Conversely, pairing 5 inescapable shocks with NE depletion by FLA-63, or both DA and NE depletion by alpha-methylparatyrosine, disrupted escape performance 24 hrs later. Residual drug effects, state dependence, or sustained amine turnover could not account for the behavioral changes. Data are discussed in terms of catecholamine mediation of escape performance through variations in response maintenance abilities. It is suggested that long-term effects of inescapable shock may be due to sensitization effects or conditioned amine depletion. (28 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Kinetics of humoral response in children with acute lymphoblastic leukemia immunized with influenza vaccine in 1993 in Poland

Songbirds exhibit seasonal changes in the volumes of song control nuclei. Birds on long, spring-like days have larger nuclei than do birds on short, winter-like days. The mechanisms mediating volumetric changes have not been determined unequivocally, but testosterone (T) is probably involved. This study examined whether testicular factors are uniquely responsible for seasonal changes in the song system, or whether photoperiod has testis-independent effects. Male American tree sparrows were exposed to one of three photoperiodic conditions: (1) Photosensitive birds were retained on short days (8L:16D). Plasma T is rarely detected in such birds. (2) Photosensitive birds were moved from short days to long days (20L:4D) and photostimulated for three weeks. Photostimulation elevates circulating T in photosensitive birds. (3) Photorefractory birds were held at least four months on 20L:4D. Such birds seldom have detectable levels of T, even though they are on long days. In each condition, there were both intact and castrated birds. Castration typically removes circulating T in tree sparrows. The volumes of the high vocal center (HVC), nucleus robustus archistriatalis (RA), and area X were measured. Song nuclei were largest in intact photostimulated birds. Other long-day birds (i.e. castrated photostimulated, and intact and castrated photorefractory groups) had larger song nuclei than did short-day intact or castrated photosensitive birds and did not differ from each other. These data indicate that photoperiod has both testis-dependent and -independent effects on the volumes of song control nuclei.     

Axonal connections of the medial magnocellular nucleus of the anterior neostriatum in zebra finches

The medial magnocellular nucleus of the anterior neostriatum (mMAN) is a small cortical nucleus which was previously identified as a component of the neural circuitry controlling vocal behavior in songbirds based on its efferent connection to the High Vocal Center (HVC), a major song control nucleus (Nottebohm et al. [1982] J. Comp. Neurol. 207:344-357; Bottjer et al. [1989] J. Comp. Neurol. 279:312-326). We have conducted tract tracing experiments (using wheat-germ agglutinin-horseradish peroxidase (WGA-HRP), the carbocyanine dye DiI, and biocytin) to determine the complete pattern of afferent and efferent connections of mMAN in adult male zebra finches. We confirmed the existence of an efferent projection from mMAN to HVC and discovered a novel projection to the region medial to caudal HVC called paraHVC (pHVC). Injections of retrograde tracers into mMAN showed that afferent input to mMAN originates from the dorsomedial nucleus of the posterior thalamus (DMP). Injections of DiI into DMP produced anterograde label over mMAN, thus confirming the DMP-to-mMAN projection. Interestingly, this anterograde label extended beyond the region of mMAN defined by HVC-projecting neurons into the immediately surrounding cortex. This extended terminal field of DMP efferents indicates that mMAN encompasses a core population of projection neurons surrounded by a shell of non-HVC-projecting neurons, both of which receive input from the dorsal thalamus. Analysis of retrograde DiI label resulting from DMP injections revealed two major sources of afferent input to DMP originating in regions of the archistriatum and hypothalamus. Inputs to DMP were distributed throughout the dorsal archistriatum and included the area that receives a projection from the parvicellular shell region of the lateral magnocellular nucleus of the anterior neostriatum, a song control nucleus, as well as the dorsal portion of the robust nucleus of the archistriatum, the motor-cortical output of the song control system. The projections from song control regions of the archistriatum to DMP may feed information back into telencephalic song control circuitry via the DMP-->mMAN-->HVC/pHVC pathway. The other source of afferent input to DMP is located in the external cellular stratum of the lateral hypothalamus (SCE). This newly delineated SCE-->DMP-->mMAN-->HVC/pHVC pathway is the first report of a hypothalamic brain region neuroanatomically integrated with song control circuitry. Because hypothalamic brain regions are important for homeostasis and regulating behavior, the trans-synaptic circuitry of mMAN may help to integrate information about the bird's internal state, such as sexual maturation, with song learning and production.