Distribution of iron in the parrot brain: conserved (pallidal) and derived (nigral) labeling patterns.

The distribution of iron in the brain of a vocal learning parrot, the budgerigar (Melopsittacus undulatus), was examined using iron histochemistry. In mammals, iron is a highly specific stain for the dorsal and ventral pallidal subdivision as well as specific cell groups in the brainstem, including the substantia nigra pars reticulata [Neuroscience 11 (1984) 595-603]. The purpose of this study was to compare the distribution of iron in the mammalian and avian brain focusing on pallidal and nigral cell groups. The results show that in the avian brain, iron stains oligodendrocytes, neurons and the neuropil. Cell staining changes dramatically along the rostrocaudal axis, with neuronal labeling confined to regions caudal to the thalamus and oligodendrocyte labeling denser in regions rostral to the dorsal thalamus. Many sensory forebrain regions contain appreciable iron labeling, including telencephalic vocal control nuclei. The dorsal and ventral subdivision of the avian pallidum, along with the basal ganglia component of the vocal control circuit, the magnicellular nucleus of the lobus parolfactorius, stain heavily for iron. Several brainstem regions, including nucleus rotundus, the medial spiriform nucleus (SpM), the principle nucleus of the trigeminal nerve, nucleus laminaris and scattered cell groups throughout the isthmus and pontine reticular formation stain intensely for iron. Within SpM neuronal labeling is more intense in the medial division while oligodendrocyte labeling is more intense in the lateral division. surprisingly no nigral iron staining was observed. Our results imply that iron is a conserved marker for the pallidum in birds and mammals, but that patterns of nigral staining have diverged in birds and mammals. Differences in iron staining patterns between birds and mammals may also reflect the relatively greater importance of the collothalamic visual pathways, pretectal-cerebellar pathways and specialized vocal learning circuitry in avian sensory and motor processing.

Projections of the oval nucleus of the hyperstriatum ventrale in the budgerigar: relationships with the auditory system.

The afferent and efferent projections of a vocal control nucleus, the oval nucleus of the hyperstriatum ventrale (HVo), were mapped out in a parrot, the budgerigar (Melopsittacus undulatus) to determine the relationships of this nucleus to the auditory system. In budgerigars, HVo is connected to both the anterior forebrain pathway as well as to nuclei forming the descending projection system to the brainstem (Durand et al. [1997] J. Comp. Neurol. 377:179-206). Previous studies (Brauth et al. [1997] Proc. N. Y. Acad. Sci. 807:368-385; Durand and Brauth [1998] Neurosci Abstr 24:78.9) indicate that HVo lesions disrupt vocal performance and that HVo neurons show long latency electrophysiologic auditory responses. HVo has also been shown to receive input from neurons in the immediately adjacent HV (Durand et al. [1997] J. Comp. Neurol. 377:179-206). Thus, the focus of the present study was to elucidate relationships between HVo, its immediately adjacent surround and telencephalic auditory nuclei. The results show that, although the lateral and medial portions of HVo are interconnected with one another, inputs to these areas and their surrounds are distinctively different. The most substantial auditory system inputs are derived from the frontal lateral neostriatum (NFl) and supracentral nucleus of the lateral neostriatum (NLs); these project primarily to the lateral HVo and lateral HVo surround. The medial HVo and surround receive only sparse or modest input from auditory nuclei, including the caudomedial neostriatum (NCM), neostriatum intermedium pars lateralis (NIL), Fields L1 and L3, and the neostriatum intermedium pars ventrolateralis (NIVL). Other sources of input to the HVo surround include the hyperstriatum accessorium (HA), the supralaminar area of the frontal neostriatum (NAs), the ventral anterior archistriatum (AAv), the medial archistriatum (Am) and the medial HV. Neurons in the HV immediately medial to HVo project to a shell region around the entire nucleus. Both the ventral paleostriatum (VP) and ventral part of the central nucleus of the lateral neostriatum (NLc) project to HVo but not to the surround. Previously described projections (Durand et al., 1997) from HVo to the NAom, NLc, and the magnicellular nucleus of the lobus parolfactorius (LPOm) were confirmed.

Distribution of tyrosine hydroxylase-containing neurons and fibers in the brain of the budgerigar (Melopsittacus undulatus): general patterns and labeling in vocal control nuclei.

The distribution of tyrosine hydroxylase (TH) was mapped out in cells and fibers of the budgerigar (Melopsittacus undulatus) brain. Special attention was given to vocal control and auditory nuclei because budgerigars are a psittacine species in which both males and females are capable of lifelong vocal learning (Farabaugh et al. [1994] J. Comp. Psychol 108:81-92). The results show that TH staining in the central nucleus of the anterior archistriatum (AAc) resembled that of surrounding archistriatal fields, except for portions of the ventral archistriatum, which exhibited substantially more TH+ fibers. Fewer fibers and fiber baskets are present in the central nucleus of the lateral neostriatum (NLc) than in surrounding fields. Both the oval nuclei of the ventral hyperstriatum (HVo) and anterior neostriatum (NAo) exhibit less fiber staining than surrounding fields whereas fiber staining in the medial NAo (NAom) and magnicellular nucleus of the parolfactory lobe (LPOm) resemble that of surrounding fields. Staining in primary telencephalic auditory nuclei was extremely low. The only sex difference observed was slightly increased TH staining in LPOm of females compared with surrounding fields on some tissue sections. These findings are in contrast to previous findings in zebra finch (Poephila guttata), a close ended vocal learning songbird in which TH staining in vocal nuclei increases during development and remains greater than surrounding fields throughout adulthood. The present results therefore support the view that catecholamines act to inhibit vocal plasticity in adult vocal learning species. Several unique features of TH-immunoreactive (ir) cell groups were observed in the brainstem including sparsely scattered TH-ir somata immediately adjacent to the third ventricle, within the tectum, basal forebrain, archistriatum, and caudal neostriatum, and in the hippocampus. These latter populations have not been described in other avian species and resemble features of the catecholamine system generally found in either reptiles or mammals.

Calcitonin gene-related peptide immunoreactive cells and fibers in forebrain vocal and auditory nuclei of the budgerigar (Melopsittacus undulatus).

The distributions of calcitonin gene-related peptide (CGRP) immunoreactive neurons and fibers were mapped within forebrain vocal control and auditory nuclei of a vocal learning psittacine species, the budgerigar (Melopsittacus undulatus). Immunoreactivity was exhibited by telencephalic nuclei previously associated with vocal control pathways on the basis of both tract tracing studies and gene mapping: the central nucleus of the anterior archistriatum (AAc), central nucleus of the lateral neostriatum (NLc), magnocellular nucleus the lobus parolfactorius (LPOm), the oval nucleus of the ventral hyperstiratum (HVo) and the medial division of the oval nucleus of the anterior neostriatum (NAom). The main body of NAo also contained an exceptionally high density of immunoreactive fibers. In contrast to the condition in oscine songbirds, CGRP-positive neuronal somata were not present in any telencephalic vocal control nucleus. CGRP-positive somata were present, however, in diencephalic cell groups that included the shell region of the nucleus ovoidalis (Ov), the nucleus dorsolateralis posterior (DLP) and a region of the ventral thalamus that was retrogradely labeled by tracer deposits into HVo and AAc. CGRP immunoreactive fibers were observed within auditory areas of the telencephalon including Field L and the neostriatum intermedium pars dorsolateralis. The likely sources of these fibers are CGRP-positive neurons within the Ov shell and DLP.

Effects of lesions of the central nucleus of the anterior archistriatum on contact call and warble song production in the budgerigar (Melopsittacus undulatus).

We studied the effects of both unilateral and bilateral lesions of the central nucleus of the anterior archistriatum (AAc) on the production of contact calls and warble song in adult male and female budgerigars. Birds were sorted into three experimental groups based on the percentage of AAc destroyed and whether lesions were unilateral or bilateral. The experimental groups were Unilateral Lesion (N = 8), Partial Bilateral Lesion (N = 5), and Bilateral Lesion birds (N = 12). Each group contained both sexes. Unilateral lesions had no demonstrable effects on contact call or warble song production. Bilateral lesions resulted in immediate and permanent disruption of all learned temporal and spectral characteristics of contact calls, although call initiation was not dependent on the AAc. Partial bilateral lesion effects varied with lesion size and location. At least 20-30% sparing of the AAc, including sparing portions of both the dorsal (AAcd) and ventral (AAcv) subdivisions on the same side of the brain, is necessary for production of prelesion contact call patterns. Warble song was absent in birds with complete bilateral destruction. Two birds with large yet incomplete lesions of the AAc sang after surgery, although the warble song of these birds was extremely impoverished and contained only a few of the typical warble song elements. Lesion results indicate that the AAc mediates the production of learned vocal features in male and female budgerigars, with each hemisphere capable of supporting a normal vocal repertoire.

Telencephalic nuclei control late but not early nestling calls in the budgerigar.

Bilateral lesions targeting the central nucleus of the anterior archistriatum (AAc) were placed in nestling budgerigars (Melopsittacus undulatus) aged 5, 9, 13, 22, 26, and 33 days post-hatch in order to evaluate the role of the telencephalon in producing nestling vocalizations in this species. In budgerigars, AAc is the final common pathway from telencephalic vocal control nuclei to brainstem respiratory and syringeal motorneuron pools. The results show that lesions destroying AAc bilaterally in addition to surrounding archistriatum and neostriatum do not alter the production of early simple patterned foodbegging calls but do prevent both the normal transition at 3-4 weeks post-hatch to more complex begging calls as well as the emergence of individually-distinctive contact calls around the time of fledging. These vocal results are strikingly similar to those obtained in previous studies in which early deafening of nestlings (Heaton and Brauth, 1999) and early lesioning of auditory areas in the anterior telencephalon (Hall WS, Brauth SE, Heaton JT. Comparison of the effects of lesions in nucleus basalis and field 'L' on vocal control learning and performance in the budgerigar (M. undulatus), Brain Behav. Evol., 1994;44:133-148) did not affect call production until 3-4 weeks post-hatch. These data combined support the idea that neither auditory feedback nor telencephalic sensorimotor circuits are necessary for the production of nestling calls before 3 weeks post-hatch.

Effects of deafening on the development of nestling and juvenile vocalizations in budgerigars (Melopsittacus undulatus).

The effects of complete and partial cochlear extirpation at ages 9-11 days posthatch were assessed in 5 nestling budgerigars (Melopsittacus undulatus) to determine if auditory feedback is necessary for the production of nestling vocalizations. Although early deafening had no effect on the production of food-begging calls produced during the first 2 weeks posthatch, deafening did disrupt the expected transition from these early calls to the longer and more complex frequency-modulated, patterned food-begging calls normally appearing 3-4 weeks posthatch. All birds sustaining either complete or partial cochlear extirpation failed to develop stereotyped contact calls around the time of fledging at 5 weeks. These results are consistent with previous research showing that deafened nestlings do not develop normal contact calls (R. J. Dooling, B. F. Gephart, P. H. Price, C. McHale, & S. E. Brauth, 1987) and also indicate that a form of sensorimotor learning is involved in the production of mature, patterned food-begging calls in budgerigars.

Cytoarchitecture of vocal control nuclei in nestling budgerigars: relationships to call development.

Changes in the cytoarchitecture of vocal control nuclei were investigated in nestling budgerigars (Melopsittacus undulatus) from hatching to fledging (five to six weeks) in relation to changes in vocalizations produced by nestlings during this period. The nuclei investigated were the hypoglossal nucleus, dorsomedial nucleus of the intercollicular midbrain, central nucleus of the archistriatum, central nucleus of the lateral neostriatum, oval nucleus of the hyperstriatum ventrale, medial division of the oval nucleus of the anterior neostriatum, and magnocellular nucleus of the lobus parolfactorius. These nuclei have been shown to form functional circuits in adults related to vocal learning. Consistent with previously reported results, we found that call development could be described in terms of five different phases based on changes in the duration and segmentation of single and multiple segment food begging calls and the appearance of the first socially learned contact calls around the time of fledging. We also found that call segment duration exhibited an inverted U-shaped developmental function during the nestling period, as has been found for total call duration. Cytoarchitectonic studies revealed striking changes in the cellular architecture of vocal control nuclei during the first four weeks posthatching. At hatching the hypoglossal nucleus exhibits adult-like cytoarchitecture, and the central nucleus of the archistriatum and the central nucleus of the lateral neostriatum are distinguishable from surrounding fields. By one week posthatch, the central nucleus of the archistriatum exhibits an adult-like appearance, while other telencephalic vocal control nuclei do not exhibit adult-like cytoarchitecture until three to four weeks posthatching. By two weeks posthatching, the dorsomedial nucleus of the intercollicular midbrain also exhibits adult-like cytoarchitecture. We observed substantial decreases in the thickness of ventricular proliferation zones during this period, with decreases in ventricular zones occurring at about the same point that nuclei at corresponding levels come to exhibit adult-like cytoarchitectonic features. Of interest is the fact that cytoarchitectural development occurs asynchronously in different brain regions, with the appearance of adult-like characteristics in the hindbrain and midbrain occurring before the appearance of adult-like cytoarchitectonic characteristics in telencephalic nuclei. These results are consistent with recent lesion studies indicating that neither auditory feedback nor telencephalic vocal control nuclei are necessary for the production of food begging and other nestling calls until three to four weeks posthatching.

Methionine enkephalin immunoreactivity in the brain of the budgerigar (Melopsittacus undulatus): similarities and differences with respect to oscine songbirds.

The brain of the budgerigar (Melopsittacus undulatus), a small parrot that acquires new vocalizations throughout life, was examined for immunoreactivity to the opioid peptide methionine enkephalin (mENK). mENK is a highly prominent feature of the chemical architecture of the forebrain vocal system of oscine songbirds. Forebrain vocal control nuclei are believed to have evolved independently in parrots and songbirds (Streidter [1994] J. Comp. Neurol. 343:35-56); however, recent studies have found similarities in the neural organization of vocal control pathways in budgerigars and songbirds (Durand et al. [1997] J. Comp. Neurol. 377:179-206). Among the similarities are the existence of recursive pathways interconnecting vocal control neurons in the archistriatum, basal ganglia (i.e., lobus parolfactorius), and dorsal thalamus. In the present study, we found that all vocal control nuclei within the budgerigar forebrain exhibit prominent mENK-like immunoreactivity (ELI) in fibers and somata. We also found striking similarities between the morphology of ELI elements in budgerigar vocal control nuclei and that described previously in songbird vocal nuclei. Despite these similarities, the budgerigar dorsal striatopallidum (lobus parolfactorius, paleostriatum augmentatum, and paleostriatum primitivum) and somatomotor (anterior) archistriatum exhibit unique patterns of ELI. The dorsal striatopallidum contained far less ELI, whereas the archistriatum contained far more than would be expected on the basis of previous studies of opioid peptides in other avian species, including pigeons, chickens, and songbirds. These differences may reflect neural specializations unique to the budgerigar that contribute to the extraordinary flexibility of the vocal motor system of this species to acquire socially significant stimuli throughout life.

Vocal control pathways through the anterior forebrain of a parrot (Melopsittacus undulatus).

A feature of the telencephalic vocal control system in the budgerigar (Melopsittacus undulatus) that has been hypothesized to represent a profound difference in organization from the oscine vocal system is its reported lack of an inherent circuit through the anterior forebrain. The present study reports anatomical connections that indicate the existence of an anterior forebrain circuit comparable in important ways to the "recursive" pathway of oscine songbirds. Results from anterograde and retrograde tracing experiments with biocytin and fluorescently labeled dextran amines indicate that the central nucleus of the anterior archistriatum (AAc) is the source of ascending projections upon the oval nuclei of the anterior neostriatum and ventral hyperstriatum (NAo and HVo, respectively). Efferent projections from the latter nuclei terminate in the lateral neostriatum afferent to AAc, thereby forming a short recurrent pathway through the pallium. Previously reported projections from HVo and NAo upon the magnocellular nucleus of the lobus parolfactorius (LPOm), and after LPOm onto the magnocellular nucleus of the dorsal thalamus (DMm; G.F. Striedter [1994] J. Comp. Neurol. 343:35-56), are confirmed. A specific projection from DMm onto NAom is also demonstrated; therefore, a recurrent pathway through the basal forebrain also exists in the budgerigar vocal system that is similar to the anterior forebrain circuit of oscine songbirds. Parallels between these circuits and mammalian basal ganglia-thalamo-cortical circuits are discussed. It is hypothesized that vocal control nuclei of the avian anterior neostriatum may perform a function similar to the primate supplemental motor area.

Functional anatomy of forebrain vocal control pathways in the budgerigar (Melopsittacus undulatus).

Budgerigars throughout life are capable of learning to produce many different sounds including those of human speech. Like humans, budgerigars use multiple craniomotor systems and coordinate both orosensory and auditory feedback in specialized forebrain nuclei. Although budgerigar auditory-vocal learning has a different evolutionary origin from that of human speech, both the human and budgerigar systems can control F0 and can alter the distribution of energy in spectral bands by adjusting the filter properties of the vocal tract. This allows budgerigars to produce an extremely diverse array of calls including many broadband and highly complex sounds.

Distribution of choline acetyltransferase and acetylcholinesterase in vocal control nuclei of the budgerigar (Melopsittacus undulatus).

The present study used histochemical methods to map the distributions of choline acetyl transferase (ChAT) and acetylcholinesterase (AChE) in the vocal control nuclei of a psittacine, the budgerigar (Melopsittacus undulatus). The distributions of ChAT and AChE in budgerigars appeared similar to that in oscine songbirds despite evidence that these systems have evolved independently. The magnicellular nucleus of the lobus parolfactorius in budgerigars, like the area X in songbirds, contained many ChAT labeled somata, fibers, and varicosities and stained densely for AChE. In contrast, the robust nucleus of the archistriatum (RA) and the supralaminar area of the frontal neostriatum in budgerigars, like the RA and the magnicellular nucleus of the neostriatum (MAN) in songbirds, respectively, contained few or no ChAT labeled somata, fibers, and varicosities and stained lightly for AChE. The central nucleus of the lateral neostriatum in budgerigars, like the higher vocal center (HVC) in songbirds, contained no ChAT labeled somata, moderate densities of ChAT labeled fibers and varicosities, and moderate levels of AChE staining. Two nuclei, the oval nucleus of the hyperstriatum ventrale (HVo) and the oval nucleus of the anterior neostriatum (NAo), contained no ChAT labeled somata, dense ChAT labeled fibers and varicosities, and moderate to high levels of AChE staining. The HVo and the NAo have no counterparts in songbirds but may be important vocal control nuclei in the budgerigar. Cholinergic enzymes are also described in other regions which may be involved in budgerigar vocal behavior, including the basal forebrain, the torus semicircularis, and the hypoglossal nuclei (nXII).

Effect of syringeal denervation in the budgerigar (Melopsittacus undulatus): the role of the syrinx in call production.

In the budgerigar, the left and right tracheosyringeal nerves (ts) were sectioned both above and below the common anastomosis in order to assess the roles of the hypoglossal nuclei and syringeal muscle halves in the control of call production. Signal processing software was used to quantify changes in contact call fundamental frequency and duration, and similarity analysis for pre- and postsurgical contact calls was performed by means of spectrogram cross-correlation. After resecting a portion of either the right or left ts nerve above the anastomosis (thereby eliminating the input from the ipsilateral half of the brainstem), contact call fundamental frequency decreased 40-50% on Day 1 postsurgery, while call structure and duration remained largely unaffected. Fundamental frequency returned to normal within 4-7 days after surgery. In contrast, nerve sectioning below the anastomosis on either side of the syrinx (thereby eliminating input to the ipsilateral half of the syringeal muscles) resulted in moderately noisy harsh-sounding calls with little change in temporal characteristics. Thus, budgerigars differ from many oscines studied to date in that they do not demonstrate laterality in vocal control at the level of the syrinx. Vocalizations produced by birds after bilateral syringeal denervation were abnormal, consisting entirely of broadband harmonic sounds with very low fundamental frequencies (i.e., less than 900 Hz) and poor frequency modulation. In contrast, individual call durations, as well as the rhythm and patterning of vocalizations resembling warble song, were remarkably similar to presurgical recordings after both unilateral and bilateral ts nerve resection.(ABSTRACT TRUNCATED AT 250 WORDS)

Covariation of binaural, concurrently-measured spontaneous otoacoustic emissions.

Simultaneous recordings of binaural spontaneous otoacoustic emissions (SOAEs) were made for 2 female subjects. For SOAEs below about 3.6 kHz measured within a testing session, the frequencies of nearby monaural and binaural SOAEs tended to move in tandem, whereas widely separated SOAEs did not. Across many testing sessions spanning a menstrual cycle, all monaural and binaural SOAE frequencies shifted in tandem. Possible mechanisms consistent with these results are discussed.

Functional anatomy of forebrain auditory pathways in the budgerigar (Melopsittacus undulatus).

Interconnections of forebrain auditory and vocal control nuclei were mapped in the budgerigar using pathway tracing techniques. The anatomical results indicate four circuits by which auditory information may influence the vocal motor system: (1) direct auditory thalamic projections from nucleus dorsomedialis posterior (DMP) to both the neostriatal higher vocal center (HVC) and robust archistriatal nucleus (RA); (2) direct projections from a neostriatal projection field of DMP (i.e., MAN, the magnocellular nucleus of the neostriatum) to HVC and RA; (3) projections from DMP and other 'accessory' auditory thalamic nuclei to the ventral paleostriatum (VP), which in turn projects to MAN and RA; (4) projections to HVC from the lateral hyperstriatum ventrale (HV), which receives input from nucleus basalis (Bas) as well as from the oval nucleus of the HV (HVo), which receives direct input from RA. Lesion methods were used to evaluate the roles of auditory pathways in call learning and production. The results show that pathways associated with Bas are essential for call production in both adult and unfledged budgerigars, while VP efferents influence vocalization only in young, unfledged budgerigars. Lesions centered in either the primary auditory neostriatum (Field L2a) or the neostriatal area in receipt of Field L input (the ventrolateral neostriatum intermedium or NIVL) did not affect vocalization in juvenile or adult budgerigars.

Comparison of the effects of lesions in nucleus basalis and field 'L' on vocal learning and performance in the budgerigar (Melopsittacus undulatus).

Lesions were placed in either nucleus basalis (Bas) or the primary thalamorecipient portion of Field 'L' (i.e. centered in Field L2a) in budgerigars at 3-5 weeks posthatching and as adults. The calls of birds sustaining Bas lesions before fledging, or as adults, were markedly abnormal in that they showed little frequency modulation and individual distinctiveness. Call durations, however, were similar for lesioned and unlesioned birds. In contrast, the calls of Field 'L' lesioned birds were similar to those of siblings and cagemates. This implies that the roles of the isthmofrontal (i.e., direct projections from the ventrolateral nucleus of the lateral lemniscus to Bas) and thalamotelencephalic (i.e., direct projections from nucleus ovoidalis thalami to Field L2a) auditory pathways in providing auditory feedback during vocal learning and performance are different and that the isthmofrontal pathway plays an essential role in these processes throughout the life of the animal.

Auditory projections to the anterior telencephalon in the budgerigar (Melopsittacus undulatus).

The connections of a higher order auditory area in the neostriatum intermedium pars ventrolateralis (NIVL) were mapped with pathway tracing techniques in order to elucidate possible pathways by which auditory feedback may influence vocal learning in the budgerigar (Melopsittacus undulatus). Previous research has shown that NIVL receives projections from Field 'L' as well as adjacent portions of the dorsolateral neostriatum intermedium (NIDL) and hyperstriatum ventrale (HV) and, therefore, may be homologous to previously described auditory centers in the dorsal and lateral portions of the auditory neostriatum of songbirds. The efferent connections of NIVL terminate within a small portion of the rostromedial archistriatum as well as a more rostrally situated area within the medial neostriatum intermedium (NI) and HV. Near by (but not overlapping) fields in NI and HV receive input from the nucleus dorsomedialis posterior thalami (DMP), the archistriatum and ectostriatum. Interestingly, only the DMP projection field overlaps a neural field known to be related to the vocal motor system. The DMP projection field corresponds to that previously described as the magnocellular nucleus of the anterior neostriatum; this nucleus is known to project to the higher vocal center in the budgerigar. In addition to projections from NIVL to medial NI and HIV, auditory information is relayed to the anterolateral telencephalon directly from the brainstem via the ventrolateral nucleus of the lateral lemniscus (VLV). This latter pathway appears comparable to that described in pigeons derived from the intermediate nucleus of the lateral lemniscus. The projection field of VLV overlaps a restricted portion of the caudal and medial aspect of nucleus basalis. These results support the notion that many aspects of telencephalic auditory pathways in birds are primitive characters, although a direct connection between auditory and vocal motor circuits was not found in the present study.

Effects of yohimbine as a reversing agent for ketamine-xylazine anesthesia in budgerigars.

Fourteen adult budgerigars (Melopsittacus undulatus) were anesthetized with a combination of ketamine hydrochloride (40 mg/kg) and xylazine hydrochloride (10 mg/kg) intramuscularly. Forty-five minutes after ketamine-xylazine injection, one of four yohimbine hydrochloride doses (0.0, 0.11, 0.275, or 0.44 mg/kg, IM) was administered in a 0.7% saline vehicle. Latencies were recorded in minutes from yohimbine injection until subjects' behavior indicated three different points of recovery: 1) lifting the head, 2) standing unaided without ataxia, and 3) perching. Means for all three recovery point latencies were significantly reduced by 0.275 mg/kg of yohimbine compared with saline vehicle alone. Mean latencies among treatment groups for each of the three recovery points were not significantly different, other than control versus treated groups. Based on these results, we recommend a yohimbine dose of 0.275 mg/kg as an effective reversing agent for ketamine-xylazine anesthesia in budgerigars.