Innate immunity and testosterone rapidly respond to acute stress, but is corticosterone at the helm?

When faced with a stressor, vertebrates can rapidly increase the secretion of glucocorticoids, which is thought to improve the chances of survival. Concurrent changes in other physiological systems, such as the reproductive endocrine or innate immune systems, have received less attention, particularly in wild vertebrates. It is often thought that glucocorticoids directly modulate immune performance during a stress response, but, in many species, androgens also rapidly respond to stress. However, to our knowledge, no study has simultaneously examined the interactions between the glucocorticoid, androgen, and innate immune responses to stress in a wild vertebrate. To address this issue, we tested the hypothesis that the change in plasma corticosterone (CORT) in response to the acute stress of capture and restraint is correlated with the concurrent changes in plasma testosterone (T) and innate immune performance (estimated by the capacity of plasma to agglutinate and lyse foreign cells) in the Abert's Towhee (Melozone aberti). Furthermore, to broaden the generality of the findings, we compared male and female towhees, as well as males from urban and non-urban populations. Acute stress increased plasma CORT, decreased plasma T in males, and decreased innate immune performance, but the increase in CORT during stress was not correlated with the corresponding decreases in either plasma T or innate immunity. By contrast, the plasma T stress response was positively correlated with the innate immune stress response. Collectively, our results challenge the proposition that the glucocorticoid stress response is correlated with the concurrent changes in plasma T, a key reproductive hormone, and innate immunity, as estimated by agglutination and lysis.

Nicotine-induced plasma corticosterone is attenuated by social interactions in male and female adolescent rats.

Most smokers begin smoking during adolescence, a period during which social reward is highly influential. Initial exposure to nicotine can produce anxiogenic effects that may be influenced by social context. This study examined play behavior and plasma corticosterone following nicotine administration (0.6 mg/kg, s.c.) in both male and female adolescent (PND39) Sprague-Dawley rats in either isolate or social contexts. In blood samples collected immediately following the 15-min test session, nicotine increased plasma corticosterone relative to saline in both male and female isolate rats, but failed to do so in both males and females placed together in same-sex pairs. Nicotine also attenuated several indices of play behavior including nape attacks, pins and social contact. In isolate rats, nicotine selectively increased locomotor activity in females; however, when administered to social pairs, nicotine decreased locomotion in both sexes. These findings suggest that the presence of a social partner may decrease the initial negative, stress-activating effects of nicotine, perhaps leading to increased nicotine reward.

Blood parasitaemia in a high latitude flexible breeder, the white-winged crossbill, Loxia leucoptera: contribution of seasonal relapse versus new inoculations.

We measured seasonal changes in the prevalence of haematozoa (Leucocytozoon fringillinarum, Haemoproteus fringillae, and Trypanosoma avium) in free-ranging White-winged Crossbills, Loxia leucoptera, over 1.5 year in Fairbanks, Alaska, USA. This prevalence was low during early winter. L. fringillinarum prevalence increased in late winter/early spring, in the absence of vectors, suggesting relapse of latent infection. By contrast, the prevalence of T. avium and H. fringillae did not increase until mid-spring, coincident with the emergence of putative vectors and suggestive of new inoculations. The winter breeding period was not associated with lower body condition or elevated blood heterophil/lymphocyte ratios than the summer post-breeding period. Thus, birds unlikely perceived their breeding effort as particularly stressful. Adult males in May and June had low plasma testosterone and their blood prevalence of L. fringillinarum, but not other haemoparasites, was higher than in adult females. This difference may have resulted from sex differences in behaviour and/or plumage colouration - bright red in males, dull green/yellow in females. Species in which reproduction and vector abundance are seasonally dissociated may constitute important models for investigating the respective contribution of reproductive hormones, breeding effort, and vector abundance to patent and latent hemoparasitic infections and to new inoculations.

Glutamatergic stimulation of luteinising hormone secretion in relatively refractory male songbirds.

Seasonal breeding in two Sonoran desert passerines, the Cassin's (Aimophila cassinii) and Rufous-crowned (Aimophila ruficeps) Sparrows is thought to be terminated by the development of a decrease in responsiveness to photostimulation, a condition known as relative photorefractoriness. It was predicted that the development of relative refractoriness is a consequence of a decrease in gonadotrophin-releasing hormone (GnRH) synthesis and associated stores of releasable GnRH. This hypothesis was tested by determining the luteinising hormone (LH) responses to the excitatory amino acid glutamate agonist N-methyl-D,L-aspartate (NMA) in males of the two species subjected to photomanipulations aimed at generating five groups: Fully photosensitive with undeveloped testes on short days (8L : 16D); fully photosensitive with developed testes on 13L : 11D; relatively photorefractory with regressed testes on 13L : 11D, and groups with developed testes held on 15L : 9D or 16L : 8D. LH release was stimulated in the Cassin's Sparrow by NMA most in the 8L group; to a lesser, but similar extent in the two 13L groups; and not at all in the 15L and 16L groups. LH release was not stimulated by NMA in any of the photoperiodic regimes in the Rufous-crowned Sparrow. In both species, NMA induced Fos-like immunoreactivity in the anterior and basal hypothalamus, but not in GnRH cell bodies. It is concluded that the development of relative photorefractoriness in Cassin's Sparrows is a consequence of reduced GnRH synthesis, reflected in a reduction in releasable GnRH. The lack of LH response of the Rufous-crowned Sparrows to NMA administration may be a consequence of high responsiveness to handling stress.

Testosterone and social context affect singing behavior but not song control region volumes in adult male songbirds in the fall.

Testosterone (T) induces singing behavior and mediates changes in the sizes and neuroanatomical characteristics of brain regions controlling singing behavior (song control regions, SCRs) in songbirds. These effects may require the enzymatic conversion of T into androgenic and estrogenic metabolites by brain tissues and can be modulated by factors such as season and social context. Testosterone administration to adult male House Finches, Carpodacus mexicanus, in the spring increases the size of their SCRs. Here, we used males of this species to investigate effects of T and T metabolism on brain morphology and singing behavior in the fall. Birds received Silastic capsules containing androgens, estrogens, and/or inhibitors of androgenic action or estrogen synthesis to determine effects of these hormones on song rates and SCR volumes. We also manipulated the social environment by changing the number of birds in visual contact with each other. Testosterone treatment stimulated singing behavior in finches held in small, visually isolated groups and exposed to song playbacks. However, administration of T or T metabolites did not increase SCR sizes. The data suggest that photoperiodic condition and social context may modulate the effects of steroids on SCRs and singing behavior.

Reproductive aging in Japanese quail, Coturnix japonica is associated with changes in central opioid receptors.

Quantitative in vitro autoradiography was used to measure specific mu and delta opioid receptor densities in regions of the Japanese quail, Coturnix japonica, brain that regulates reproductive endocrine and behavioral responses to determine the possible involvement of the opioid system in reproductive decline seen during aging. Densities were measured in selected brain regions of young sexually active (YAM), young photoregressed (YPM), old reproductively senescent (OIM) male, young active (YF), and old senescent female (OF) Japanese quail. Medial and lateral septum (SM, SL), medial preoptic area (POM), and n. intercollicularis (ICo) were of particular interest for reproductive responses. Similar to previous observations, mu and delta opioid receptors showed differential distributions in the areas measured. Some age-related changes were observed, with lower SM mu receptor densities in aged males (OIM) than females or young males (YAM). Densities of mu receptors in the POM and in other areas examined did not vary with sex or age. Similarly, OIM males had lower densities of delta receptors in the SM than young males (YAM and YPM); POM delta receptor densities were also low in OIM males compared to the YPM males, and YAM males were intermediate. Interestingly, photoregressed males (YPM) had higher SL delta receptor densities than any other group. Thus there were age-related differences detected in mu receptor densities among groups in the SM of OIM relative to other groups; and the mu and delta receptor densities did not differ in females with brain region. Additionally for delta receptors specifically, YF and OF did not differ from OIM for any brain region and similarly had lower densities of delta receptors compared to YAM males. These data provide support for regional differences in opioid receptor distribution and for age- and sex-related differences in delta opioid receptor densities. The direction of change presents an interesting dichotomy in that, compared to young active males, delta opioid receptor densities increased with loss of reproductive function in the YPM, whereas receptor densities decreased in the OIM. Plasma androgen levels were relatively low in both these groups compared to the young active males. This observation suggests that there is an age-related loss in the ability of this receptor system to respond to circulating and centrally produced steroid hormones in the POM and in some septal regions, compared to young animals that are responding to environmental cues. Furthermore, these data support an active role of the opioid peptide system in the inhibition of the reproductive axis in photoregression.

Region-specific testosterone modulation of the vasotocin-immunoreactive system in male dark-eyed junco, Junco hyemalis.

The nonapeptide vasotocin (VT) is the avian equivalent of the mammalian antidiuretic hormone vasopressin and is believed to control aggressive and reproductive behaviors. Brain VT distribution has been described in several domesticated avian species. We previously demonstrated that VT distribution in the brain of a free-ranging male passerine, the dark-eyed Junco, Junco hyemalis, resembles that in domesticated birds. A preliminary study also suggested that the VT-immunoreactive (VT-ir) system of juncos is regulated by testosterone (T), as is the case of galliforms. To test this hypothesis, we investigated the effects of castration and T replacement on brain VT-ir innervation in adult male juncos. Castration reduced VT-ir innervation in the lateral septum (SL), the medial preoptic nucleus, the nucleus of the stria terminalis and the intercollicularis nucleus. These effects of castration were largely reversed by T treatment at high physiological doses, but significantly so only for the SL. Given the demonstrated behavioral role of the above VT-ir-containing brain regions, the results suggest that these regions may be sites of action of VT on reproductive behaviors.

Reproductive endocrinology of a free-living, opportunistically breeding passerine (white-winged crossbill, Loxia leucoptera).

The aim of the study was to investigate, in a free-living opportunistic breeder, the White-winged Crossbill (Loxia leucoptera), the degree to which annual changes in photoperiod and differences in the timing of breeding in different years correlate with the secretion of reproductive hormones. Seasonal changes in concentrations of luteinizing hormone (LH) and prolactin were compared in White-winged Crossbills in interior Alaska (64 degrees 50'N, 147 degrees 50'W) in plasma samples taken in May-December 1994 and July 1998-August 1999. The birds were in breeding condition in May-July 1994 and in February-April 1999. Additional observations were made on plasma testosterone (T) in males in 1998/1999. In 1998, in adult males, plasma LH and T were low in September-October (LH: <0.4 ng/ml; T: undetectable) and increased significantly in November (LH: 3.65 +/- 2.20 ng/ml) and December (T: 0.69 +/- 0.88 ng/ml). Thereafter, plasma T increased to 2.35 +/- 2.03 ng/ml in March/April and returned to basal levels by June/July. In 1998/1999, in females, plasma LH did not change seasonally. In males in 1994, plasma LH was higher in June (2.15 +/- 1.22 ng/ml) and July (2.86 +/- 0.69 ng/ml) than in the same period in 1999 (June: 1.20 +/- 1.82 ng/ml; July: 1.37 +/- 1.03 ng/ml), which is consistent with the later breeding season in 1994. In 1994 and 1998/1999 in both sexes, concentrations of plasma prolactin were elevated when day lengths exceeded about 12 h, being high during late spring to early fall (males: 40-130 ng/ml; females: 40-140 ng/ml) and low during winter (<15 ng/ml in both sexes). In 1994 and 1998, molt occurred during the fall, at a time when breeding did not occur, although conifer seeds were sufficiently abundant to support a large bird population. These data are consistent with the view that White-winged Crossbills have the potential to breed at all times of the year except when molting in the fall. It is likely that a photoinduced increase in prolactin secretion is a factor that induces molt. Crossbills may not breed when they are molting because of increased metabolic demands for feather formation and for the maintenance of basal metabolism at a time of year when ambient temperatures fall well below 0 degrees. Alternatively or in addition, the birds may become reproductively photorefractory.

Testosterone, corticosterone, and photoperiod interact to regulate plasma levels of binding globulin and free steroid hormone in dark-eyed juncos, Junco hyemalis.

The pharmacology and regulation of corticosteroid binding globulins (CBG) in Dark-eyed Juncos, Junco hyemalis, was investigated. The equilibrium dissociation constant for [(3)H]corticosterone (CORT) binding to plasma was <5 nM. This binding site had a similar high affinity for progesterone, approximately fivefold lower affinity for androgens, and negligible affinity for estradiol. The following data suggested that plasma CBG levels are regulated by both testosterone and day length: (1) CBG binding capacity in free-living adult males was greater in early than in late breeding season and greater in males than in females and (2) CBG levels were higher in testosterone-treated, castrated males than in castrated males receiving no testosterone and still higher in testosterone-treated males exposed to long days than in similar males exposed to short days. Birds apparently lack a sex steroid-specific binding globulin, but it was estimated that more than 90% of testosterone in junco plasma should bind to CBG. An increase in plasma CORT, such as occurs during a stress response, was judged to acutely increase free testosterone levels as much as fivefold. Corticosterone and testosterone may thus interact in a complex manner in species that lack sex hormone binding proteins.

Seasonal and age-related changes in blood parasite prevalence in Dark-eyed Juncos (Junco hyemalis, Aves, Passeriformes).

We determined seasonal changes in blood parasite infections in a free-living population of Dark-eyed Juncos (Junco hyemalis) breeding in interior Alaska (65 degrees N; 148 degrees W). The common parasites found in blood smears were Leucocytozoon fringillinarum (56%), Trypanosoma avium (33%), and Haemoproteus fringillae (9%). In males, parasite prevalences were relatively high at arrival on breeding grounds and increased during the breeding season. Intensity of infection with Leucocytozoon also increased between spring and summer, and then decreased at the time of migration (September). This decrease did not occur in adult females. Elevated prevalences during the breeding season probably reflected the addition of new cases via vector activity to positive status resulting from spring relapse. We observed neither an association between parasite species nor a consistent relationship between parasite intensity and body condition. To further study relationships between reproductive system activity and parasite infections, we compared prevalences in adult males that were undergoing their first cycle of gonadal development and regression (males in their second calendar year, or SY) with those of older males (males in their third or more calendar year, i.e., after-second-year males or ASY). Circulating testosterone concentrations declined in both groups between arrival on breeding grounds (end of April-early May) and the end of the reproductive period (July), and they were higher in May in ASY than in SY males. At the peak of the breeding season (June), ASY males also had a higher parasite prevalence than SY males. This difference may have resulted from immunosuppressive effects of gonadal hormones and/or from behavioral differences between SY and ASY males such that older males were exposed to more insect vectors than younger males. .

Interspecific variability of prevalence in blood parasites of adult passerine birds during the breeding season in Alaska.

Blood parasite prevalence based on microscopic examination of stained blood smears was determined in adults of 11 passerine bird species sampled during their breeding season (May and June 1997-98) in interior Alaska (USA). These species included primarily Nearctic migratory species such as the dark-eyed junco (Junco hyemalis) and neotropical migratory species such as the blackpoll warbler (Dendroica striata), alder flycatcher (Empidonax alnorum), Swainson's thrush (Catharus ustulatus), northern waterthrush (Seiurus noveboracensis), and bank swallow (Riparia riparia) as well as one long-distance palearctic migrant, the arctic warbler (Phylloscopus borealis). The more prevalent parasites were Leucocytozoon dubreuili (73% of the sampled turdinids), L. fringillinarum (42% of the sampled fringillids and parulids), and Trypanosoma avium (39% of the sampled hosts). Other parasites (H. fallisi: 18% of the sampled turdinids; Haemoproteus paruli: 14% of the sampled parulids; H. fringillae: 5% of the sampled fringillids; microfilariae: 4% of the sampled hosts) were observed less frequently. Plasmodium vaughani was found only in two yellow warblers (Dendroica petechia). Overall parasite prevalence varied between 0% in the alder flycatcher to >80% in Swainson's thrush, arctic warbler, and Townsend's warbler (Dendroica townsendi). Prevalence of various hematozoa also was bird species-dependent. No relationship was observed between prevalence and either foraging (aerial versus trees/shrubs) or nesting habits (ground versus arboreal) or general location of the wintering area of the different species examined. Prevalence also was unrelated to average dates of arrival on breeding grounds and, therefore, to potential duration of exposure to local insect vectors before capture. Differences in blood parasite prevalence among species breeding in a same region and in the same type of habitat may result from differences in host specificity such as immunological resistance to infection or blood meal preference by potential vectors and/or in behavioral adjustments/physiological traits that alter exposure to vectors.

Effects of testosterone and photoperiodic condition on song production and vocal control region volumes in adult male dark-eyed juncos (Junco hyemalis).

In seasonally breeding male oscines, song learning and expression are controlled by brain regions (vocal control regions, VCRs) that exhibit seasonal neural plasticity in adulthood. Several VCRs contain androgen receptors, and gonadal androgens play important roles in the control of seasonal structural and functional changes of VCRs. Recent studies also found that adult VCRs are influenced by factors other than gonadal hormones, including photoperiod, but the relative importance of these factors and their mechanisms of action are poorly understood. To address this issue, we investigated the contributions of photoperiod and testicular androgens to the regulation of VCR volumes and to the control of song expression in adult dark-eyed juncos, Junco hyemalis. Exposing castrated (CX) photosensitive males to long days (LD) enhanced their high vocal center (HVc) volumes compared to those of males held on short days (SD). These volumes were not further increased by concurrent testosterone (T) treatment, revealing a marked and gonadal androgen-independent stimulatory influence of photoperiod on the size of this brain region. HVc sizes were smaller in LD-exposed photorefractory than photosensitive males irrespective of whether birds were intact or had been castrated before photoperiodic manipulations, but HVc sizes increased in response to T treatment in intact photorefractory males. Thus, LD exposure can increase HVc volumes in the absence of gonadal T, but large volume induction in photorefractory males requires elevated plasma T levels. Testosterone treatment of SD-exposed photosensitive males increased HVc, but not Area X, MAN, or RA volumes. Only T-treated males sang and this treatment given to castrates was equally effective behaviorally when administered to photosensitive, photostimulated, or photorefractory juncos. This result indicates that the stimulating influence of LD exposure on HVc volumes is insufficient to induce song in the absence of elevated plasma T levels.

Seasonal changes in brain GnRH immunoreactivity and song-control nuclei volumes in an opportunistically breeding songbird.

White-winged crossbills (Loxia leucoptera) are opportunistic breeders that can nest at almost any time of year if there is sufficient food. Other cardueline finches that have been shown to breed on a strictly seasonal schedule become absolutely refractory to the stimulatory effects of long-day photoperiod, dramatically down-regulate hypothalamic gonadotropin-releasing hormone (GnRH), and reduce the volume of several song-control nuclei in autumn. This study examined whether changes in photoperiod modify the GnRH and song-control systems in white-winged crossbills. Adult male and female crossbills were captured and held on a naturally changing photoperiod. Brains of male and female birds were collected in May, October, and January. GnRH content was assessed by immunocytochemistry and the volumes of Nissl-defined song-control nuclei (HVc, Area X, and the robust nucleus of the archistriatum) were reconstructed. In contrast to other cardueline finches, GnRH immunoreactivity was relatively stable across the year, exhibiting only modest seasonal variation. The song control system, on the other hand, exhibited large seasonal changes as well as sex differences. Thus, crossbills appear to maintain hypothalamic GnRH content year round, perhaps to facilitate a rapid response to favorable breeding conditions, even on short days. However, song control nuclei are dramatically affected by photoperiod. Future work should examine these systems in crossbills breeding on short days to compare photoperiod-dependent and -independent effects on neural plasticity.

Year-class differences in the reproductive system, plasma prolactin and corticosterone concentrations, and onset of prebasic molt in male dark-eyed juncos (Junco hyemalis) during the breeding period.

Year-class differences in reproductive function were investigated in a free-living population of adult male Dark-eyed Juncos, Junco hyemalis, breeding in interior Alaska. Second-year males (SY, entering their first breeding season) were compared with after-second-year males (ASY, entering at least their second breeding season). We measured body mass, size of the cloacal protuberance (CP), testis mass, onset of prebasic molt, and concentrations of plasma luteinizing hormone (LH), testosterone (T), corticosterone (CORT), and prolactin (PRL) throughout the reproductive season (April to mid-July). There were no differences in SY and ASY body weights but SY males had smaller CPs and testis masses than ASY males during gonadal recrudescence and at the end of the breeding season. Plasma LH was elevated from April until mid-June and then decreased in the same way in both year classes. In contrast, plasma T was high from April until mid-May and was lower in SY than in ASY juncos shortly after they arrived on their breeding grounds at the end of April, but not at other times. In July, SY males started to molt earlier, suggesting that they became photorefractory earlier than ASY males. Plasma PRL increased progressively in both year classes between April and early June and decreased in early July. At this time, plasma PRL decreased earlier in SY than in ASY males. Plasma CORT changed seasonally, but did not differ between SY and ASY juncos. Thus, year-class differences in CP sizes and testis mass apparently did not result from SY males secreting less LH or more PRL or CORT than ASY males. It is suggested that differences in reproductive condition in SY and ASY juncos are mediated by interactions with conspecific birds and do not result from an intrinsic effect of age.

Vocal control region sizes of an adult female songbird change seasonally in the absence of detectable circulating testosterone concentrations.

Previous research established that in several species of seasonally breeding oscine birds, brain areas [vocal control regions (VCRs)] that control vocal behavior learning and expression exhibit seasonal plasticity, being larger during than outside the reproductive period. In adult males, this seasonal decrease correlates with circulating testosterone (T) concentrations. VCRs contain androgen receptors and T plays an important role in neural plasticity and in the control of singing behavior. In behaviorally dimorphic species, VCRs are larger in males than females and change seasonally also in females, but the dependency of these changes on circulating T levels in females has not been established. In free-living adult dark-eyed juncos (Junco hyemalis), a species in which females do not normally sing, the sizes of three VCRs (high vocal center, robust nucleus of the archistriatum, and Area X) were larger in males than females and decreased between summer and fall in both sexes. In males, this decrease was associated with changes in circulating T concentrations. Females, however, had on average undetectable T levels throughout the breeding season. Seasonal changes in VCR volumes in adult females may depend on very low (below detection limit) circulating T concentrations, on nonandrogenic plasma steroids, on androgen (or androgen metabolites) produced in brain tissues, and/or on nonsteroidal factors such as photoperiod or social interactions with conspecific birds.

Changes in brain gonadotropin-releasing hormone- and vasoactive intestinal polypeptide-like immunoreactivity accompanying reestablishment of photosensitivity in male dark-eyed juncos (Junco hyemalis).

In seasonally breeding, photoperiodic birds, the development of photorefractoriness is associated with decreased brain expression of gonadotropin-releasing hormone-like immunoreactivity (GnRH-li ir) and increased expression of vasoactive intestinal polypeptide-like immunoreactivity (VIP-li ir). Dissipation of photorefractoriness and reestablishment of photosensitivity are associated with increased GnRH-li ir brain production, but concurrent changes in VIP-li ir expression have not been investigated. To address this question, we compared the expression of VIP-li ir in the infundibulum (INF) of adult male dark-eyed juncos (Junco hyemalis) that were made photorefractory (PR) by prolonged exposure to long days with that of birds that were not photostimulated (PS), but had regained photosensitivity by exposure to short days for 5 (short-term-PS, ST-PS) or 13 (long-term-PS, LT-PS) consecutive months. Photosensitive males had smaller INF VIP-li ir cell bodies than PR males, but the numbers of INF VIP-li ir cells were independent of photoperiodic condition. Changes in infundibular VIP-li ir were correlated with changes in preoptic area (POA) GnRH-li expression. Specifically, photosensitive males had more and larger POA GnRH-li ir cells and more GnRH-li ir fibers in this region than PR males. Further, LT-PS males had more GnRH-li ir POA fibers and larger testes than ST-PS juncos. Thus, induction of photorefractoriness is associated with increased VIP and decreased GnRH brain expression whereas dissipation of photorefractoriness concurs with decreased VIP and increased GnRH brain expression. These results suggest a physiological role for VIP in the control of changes in GnRH expression as a function of the photosensitive condition.

Central vasotocin-immunoreactive system in a male passerine bird (Junco hyemalis).

Previous investigations have identified regions of the avian brain that contain immunoreactive vasotocinergic (VT-ir) cell bodies and fibers. These studies exclusively used domesticated species, and the relevance of the findings for free-living birds has not been established. The present study used immunocytochemistry to determine the neuroanatomical distribution of the VT-ir system in the brain of a well-studied male passerine bird (dark-eyed junco, Junco hyemalis) obtained from a natural population in interior Alaska (65 degrees N, 147 degrees W). VT-ir cell bodies were observed in several brain regions (paraventricular and supraoptic nuclei, nucleus of the stria terminalis), where they have been described in other oscine species. VT-ir fibers were widespread in many brain regions and were especially abundant in the medial preoptic nucleus, the basal region of the septum, and the hypothalamic-neurohypophyseal tract. Fibers were also present in brain regions that are involved in the control of vocal behavior including the ventromedial capsular region of the nucleus robustus archistriatalis and the dorsomedial portion of the mesencephalic nucleus intercollicularis. The widespread brain distribution of VT-ir cell bodies and fibers in juncos generally resembles that of domestic birds and suggests a role for this neuropeptide in the control of reproductive behavior and physiology.

Age- and sex-related differences in opioid receptor densities in the songbird vocal control system.

Avian vocal control regions of adult male songbirds contain opioid peptides and receptors, suggesting that opioids play a role in avian vocal behavior control. In a previous study, we found no difference in opioid receptor densities in singing versus nonsinging adult male dark-eyed juncos (Junco hyemalis), leading us to hypothesize that opioids are not involved in controlling song production. To assess whether opioids may be involved in other aspects of vocal behavior, we used quantitative in vitro autoradiography to compare mu and delta opioid receptor densities in vocal control regions of singing adult males with those of adult females and adolescent (about 3 months old) males and females. We found mu and delta receptors in all vocal control regions measured. Adolescents had significantly higher opioid receptor densities than did adults in area X (delta), robust n. of the archistriatum (delta and mu), and n. intercollicularis (mu), suggesting a developmental role for opioids in the vocal control system. Based on opioid roles in other animal models, we propose that opioids may be involved in song learning, auditory processing, and/or vocal control system development.

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.

Affinity of naloxone and its quaternary analogue for avian central delta and mu opioid receptors.

Quaternary narcotic antagonists that are assumed not to penetrate the blood-brain barrier following systemic administration are commonly used to distinguish between peripheral and central actions of opiates. In mammals, these antagonists have a lower affinity for opioid receptors than their tertiary parent compounds. The relative affinity of quaternary vs. tertiary antagonists either for opioid receptors in non-mammalian species or for specific receptor subtypes has, however, not been determined. Using brain tissues from a passerine songbird (Junco hyemalis), we found the affinity of the quaternary antagonist, naloxone methiodide (Nal MI), for brain opioid receptors to be less than 10% that of Nal HCl. Further, Nal MI affinity for mu and delta receptors is 8.7% and 3.7%, respectively, that of Nal HCl. These results confirm that tertiary narcotic antagonist quaternization substantially reduces the affinity of these derivatives for central opioid receptors. They show that this reduction is receptor-type selective, and they extend previous reports demonstrating functional similarities between mammalian and non-mammalian central opioid receptors.