Nucleus Intercollicularis Research Articles

The neural distribution of the avian homologue of oxytocin, mesotocin, in two songbird species, the zebra finch and the canary: A potential role in song perception and production.

The avian homologue of oxytocin (OT), formerly called mesotocin, influences social behaviors in songbirds and potentially song production. We sought to characterize the distribution of OT peptide in the brain of two songbird species: canaries (Serinus canaria) and zebra finches (Taeniopygia guttata). To visualize OT, we performed immunocytochemistry using an antibody previously shown to identify OT in avian species. In both canaries and zebra finches, dense OT‐ir perikarya were located in the paraventricular nucleus (PVN), preoptic area (POA), supraoptic nucleus (SON), and medial bed nucleus of the stria terminalis (BNSTm). We also observed morphologically distinct OT‐ir cells scattered throughout the mesopallium. OT‐ir fibers were observed in the PVN, ventral medial hypothalamus (VMH), periaqueductal gray (PAG), intercollicular nucleus (ICo), and ventral tegmental area (VTA). We also observed punctate OT‐ir fibers in the song control nucleus HVC. In both male and female canaries, OT‐ir fibers were present in the lateral septum (LS), but innervation was greater in males. We did not observe this sex difference in zebra finches. Much of the OT staining observed is consistent with general distributions within the vertebrate hypothalamus, indicating a possible conserved function. However, some extra‐hypothalamic distributions, such as perikarya in the mesopallium, may be specific to songbirds and play a role in song perception and production. The presence of OT‐ir fibers in HVC and song control nuclei projecting dopaminergic regions provides anatomical evidence in support of the idea that OT can influence singing behavior—either directly via HVC or indirectly via the PAG, VTA, or POA.

Investigation of stereotactic surgery for avian brain stimulation by a fully implanted wireless system.

The authors' goal was to study avian motor brain mapping via wireless stimulation to induce certain behaviors. In this paper, the authors propose an electrode design that is suitable for avian brain stimulation as well as a stereotactic implant procedure for the proposed electrode. An appropriate breed for avian brain study was chosen. A fully implantable remote-controlled electrical stimulation system was inserted to minimize discomfort. A suitable electrode design and stereotactic surgery method based on the electrode design were investigated. Using a wireless stimulation system, flapping and rotation behaviors were induced by stimulating the ventral part of the nucleus intercollicularis and formatio reticularis medialis mesencephali both on the ground and during flight. The authors were able to implant the entire brain stimulation system inside the avian body without any surgical complications. Postoperative observations suggested that the bird did not find the implant uncomfortable.

Cholecystokinin induces crowing in chickens

Animals that communicate using sound are found throughout the animal kingdom. Interestingly, in contrast to human vocal learning, most animals can produce species-specific patterns of vocalization without learning them from their parents. This phenomenon is called innate vocalization. The underlying molecular basis of both vocal learning in humans and innate vocalization in animals remains unknown. The crowing of a rooster is also innately controlled, and the upstream center is thought to be localized in the nucleus intercollicularis (ICo) of the midbrain. Here, we show that the cholecystokinin B receptor (CCKBR) is a regulatory gene involved in inducing crowing in roosters. Crowing is known to be a testosterone (T)-dependent behavior, and it follows that roosters crow but not hens. Similarly, T-administration induces chicks to crow. By using RNA-sequencing to compare gene expression in the ICo between the two comparison groups that either crow or do not crow, we found that CCKBR expression was upregulated in T-containing groups. The expression of CCKBR and its ligand, cholecystokinin (CCK), a neurotransmitter, was observed in the ICo. We also showed that crowing was induced by intracerebroventricular administration of an agonist specific for CCKBR. Our findings therefore suggest that the CCK system induces innate vocalization in roosters.

Locomotion Control of Pigeons using Polymer-based Deep Brain Electrodes.

This paper describes the electrical modulation of locomotion in pigeons using deep brain electrodes. Polymer-based depth electrodes with four channels were fabricated. Based on the location of the nucleus intercollicularis (ICo), the shanks of the depth electrodes were designed to be a length of 11 mm. After the implantation of the depth electrode into the ICo region of the brain, it was connected by wires to a custom-made stimulator, and biphasic current pulses were delivered. Current pulses with an amplitude of 0.5 mA, a rate of 58.0 Hz, and a duration of $320\mu \mathrm{s} $s were applied for 0.5 s. When the ICo region was electrically stimulated, taking-off behavior was successfully induced for 0.4 s. Induction of taking-off behavior by electrical stimulation, when coupled to control of turning and running forward locomotions, may contribute to the development of remote flight-control system of freely moving pigeon.

Intercollicular nucleus electric stimulation encoded “walk forward” commands in pigeons

Abstract The bio-robot research field is growing. Robo-pigeons have been successfully programmed to turn left or right; however, a satisfactory method of commanding a robo-pigeon to walk forward is still lacking. This problem has become a roadblock to progress in bio-robot research and applications. In mammals, the midbrain periaqueductal gray region (PAG) plays a key role in mediating defensive reactions in response to fear and anxiety. The avian intercollicular nucleus (ICo) is thought to correspond to the PAG. In this study, we found that microstimulating the ICo could successfully induce a robo-pigeon to walk forward. Compared with stimulation of the previously used archistriatum, the response time was considerably shorter and the behavior accuracy significantly higher. This paper describes in detail the process of controlling a robo-pigeon such that it walks forward and backward along a prescribed straight line. From the results, we draw the conclusion that the ICo is suitable for prompting the “walk forward” order in robo-pigeons.

Heterogeneous organization and connectivity of the chicken auditory thalamus (Gallus gallus).

The auditory ascending system contains parallel pathways in vertebrate brains. In chickens (Gallus gallus), three pathways arise from nucleus laminaris (NL), nucleus angularis (NA), and regio intermedius (RI) in the brainstem, innervating three subdivisions of the nucleus mesencephalicus lateralis pars dorsalis (MLd) in the midbrain. The current study reveals the segregation of these pathways in their subsequent projections to the nucleus ovoidalis (Ov) in the thalamus. Based on cytoarchitecture and myelin distribution, we identified seven Ov subregions, including five neuronal clusters within the Ov proper, the nucleus semilunaris parovoidalis (SPO), and the circum-ovoidalis (cOv). Immunocytochemistry further revealed that a ventromedial cluster of the Ov proper (Ovvm) contains unique cell types expressing α8 subunit nicotinic acetylcholine receptor, while SPO and cOv are characterized with expression of calcitonin-gene-related peptide and cholecystokinin. Tract tracing studies demonstrated that Ovvm is a major target of the NL-recipient zone of MLd, while the RI-recipient zone of MLd predominantly projects to a ventrolateral cluster of the Ov proper. Afferent inputs to the remaining regions of the Ov proper mostly arise from the NA-recipient zone of MLd. SPO and cOv receive a projection from the surrounding areas of MLd, named the nucleus intercollicularis. Importantly, the Ov proper, SPO and cOv all project to the Field L2 in the forebrain, the avian auditory cortex. Taken together, these results demonstrate that the avian auditory thalamus is a structurally and functionally heterogeneous structure, implicating an important role in generating novel representations for specific acoustic features.

The ventromedial hypothalamic nucleus in the zebra finch (Taeniopygia guttata): Afferent and efferent projections in relation to the control of reproductive behavior.

Sex-specific mating behaviors occur in a variety of mammals, with the medial preoptic nucleus (POM) and the ventromedial hypothalamic nucleus (VMH) mediating control of male and female sexual behavior, respectively. In birds, likewise, POM is predominantly involved in the control of male reproductive behavior, but the degree to which VMH is involved in female reproductive behavior is unclear. Here, in male and female zebra finches, a combination of aromatase immunohistochemistry and conventional tract tracing facilitated the definition of two separate but adjacent nuclei in the basal hypothalamus: an oblique band of aromatase-positive (AR+) neurons, and ventromedial to this, an ovoid, aromatase-negative (AR-) nucleus. The AR- nucleus, but not the AR+ nucleus, was here shown to receive a projection from rostral parts of the thalamic auditory nucleus ovoidalis and from the nucleus of the tractus ovoidalis. The AR- nucleus also receives an overlapping, major projection from previously uncharted regions of the medial arcopallium and a minor projection from the caudomedial nidopallium. Both the AR- and the AR+ nuclei project to the intercollicular nucleus of the midbrain. No obvious sex differences in either the pattern of AR immunoreactivity or of the afferent projections to the AR- nucleus were observed. The significance of these results in terms of the acoustic control of avian reproductive behavior is discussed, and a comparison with the organization of VMH afferents in lizards suggests a homologous similarity of the caudal telencephalon in sauropsids.

Dorsal pallidal neurons directly link the nidopallium and midbrain in the zebra finch (Taeniopygia guttata).

The dorsal pallidum in birds is considered similar, if not homologous, to the globus pallidus (GP) of mammals. The dorsal pallidum projects to both thalamic and midbrain targets similar to the direct and indirect pathways arising from the internal and external segments of the GP. In the present study, retrograde and anterograde tracing studies revealed a previously undescribed projection of the avian dorsal pallidum. This arises from a specific dorsomedial component, which terminates in the intercollicular nucleus and partly surrounds the avian equivalent of the central nucleus of the inferior colliculus. The respiratory-vocal dorsomedial nucleus of the intercollicular complex, however, does not receive these projections. The somata of the pallidal neurons retrogradely labeled from injections in the intercollicular nucleus were large and generally multipolar and had extensive, sparsely branching central processes (presumptive dendrites) that together extended up to 2 mm dorsally into the intermediate and caudomedial nidopallium. The size and morphology of these neurons were similar to those of large pallidal neurons labeled by calretinin immunoreactivity, which could be co-localized to the same cells. Thus, rather than being directly involved in the control of movement, the large dorsomedial neurons of the caudal dorsal pallidum may be involved in sensory processing, in that they provide an unusual direct link between sensory (auditory/somatosensory) regions of the nidopallium and sensory regions of the intercollicular nucleus of the midbrain. J. Comp. Neurol. 525:1731-1742, 2017. © 2016 Wiley Periodicals, Inc.

The mesencephalic GCt-ICo complex and tonic immobility in pigeons (Columba livia): a c-Fos study.

Tonic immobility (TI) is a response to a predator attack, or other inescapable danger, characterized by immobility, analgesia and unresponsiveness to external stimuli. In mammals, the periaqueductal gray (PAG) and deep tectal regions control the expression of TI as well as other defensive behaviors. In birds, little is known about the mesencephalic circuitry involved in the control of TI. Here, adult pigeons (both sex, n=4/group), randomly assigned to non-handled, handled or TI groups, were killed 90min after manipulations and the brains processed for detection of c-Fos immunoreactive cells (c-Fos-ir, marker for neural activity) in the mesencephalic central gray (GCt) and the adjacent nucleus intercollicularis (ICo). The NADPH-diaphorase staining delineated the boundaries of the sub nuclei in the ICo-GCt complex. Compared to non-handled, TI (but not handling) induced c-Fos-ir in NADPH-diaphorase-rich and -poor regions. After TI, the number of c-Fos-ir increased in the caudal and intermediate areas of the ICo (but not in the GCt), throughout the rostrocaudal axis of the dorsal stratum griseum periventriculare (SGPd) of the optic tectum and in the n. mesencephalicus lateralis pars dorsalis (MLd), which is part of the ascending auditory pathway. These data suggest that inescapable threatening stimuli such as TI may recruit neurons in discrete areas of ICo-GCt complex, deep tectal layer and in ascending auditory circuits that may control the expression of defensive behaviors in pigeons. Additionally, data indicate that the contiguous deep tectal SCPd (but not GCt) in birds may be functionally comparable to the mammalian dorsal PAG.

Albert Feng and target ranging by echolocating bats

Ultrasonic biosonar sounds propagate out to a target and return as echoes. Simultaneously, phasic-on neural responses evoked by each outbound sound propagate from one subpopulation of neurons to another at different latencies in the inferior colliculus, creating delay-lines carrying a replica of the broadcast. When each echo returns, it evokes new on-responses that travel over similar subpopulations of neurons, but lagging behind responses to the broadcast according to the delay of the echo. Albert Feng’s discovery of delay-tuned neurons in the bat’s nucleus intercollicularis revealed that bats “read out” echo delay from the delay-lines by detecting the succession of coincidences between responses to the echo and to the emission. Subsequent work in multiple labs demonstrated the ubiquity of delay-tuning in bats. Working in the inferior colliculus, Al Feng showed how the delay-line responses are restricted to single spikes by initial inhibition that determines each neuron’s characteristic on-response latency followed by new inhibition that suppresses any subsequent multiple spikes. The interplay between timed and triggered inhibition defines the delay-lines, while delay-tuned coincidence-detecting neurons from inferior colliculus to superior colliculus guide the bat and from inferior colliculus to auditory cortex create the bat’s perceived images. [Work supported by ONR.]

Glutamatergic circuits in the song system of Zebra Finch brain determined by gene expression of Vglut2 and Glutamate receptors

The songbird brain has a system of interconnected nuclei that are specialized for singing and song learning. Electrophysiological findings indicate a role for the glutamatergic neurons in the song system. Vesicular glutamate transporter 2 (vGluT2) is considered to be a specific biomarker of glutamatergic neurons in birds. Neurons receiving glutamatergic afferents express mRNA of ionotropic glutamate receptor subunits. This study examined expression of vGluT2 and glutamate receptor subunit mRNAs in nuclei of the song pathways of male zebra finch brain by in situ hybridization. VGluT2 mRNA was revealed high density of expression in the song nuclei, namely HVC, lateral magnocellular nucleus of the anterior nidopallium, and robust nucleus of the arcopallium. Area X did not show expression of vGluT2 mRNA. Nuclei in the descending motor pathway (dorsomedial nucleus of the intercollicular complex and retroambigual nucleus) were expressed vGluT2 mRNA. Target nuclei of vGluT2 mRNA-expressing nuclei showed hybridization signals for mRNAs of ionotropic glutamate receptor subunits. At least one of five subunit mRNAs (GluA1, GluA4, GluK1, GluN1, GluN2A) was expressed in song nuclei. The present findings support the existence of glutamatergic circuits in the song system in songbirds. DOI: http://dx.doi.org/10.3329/ralf.v1i1.22356 Res. Agric., Livest. Fish.1(1): 61-70, Dec 2014

Nesting behavior is associated with VIP expression and VIP–Fos colocalization in a network-wide manner

Many species, including humans, engage in a series of behaviors that are preparatory to the arrival of offspring. Such “nesting behaviors” are of obvious importance, but relevant neuroendocrine mechanisms remain little studied. We here focus on the potential roles of vasoactive intestinal polypeptide (VIP) in the performance of appetitive and consummatory nesting behaviors in male and female zebra finches (Taeniopygia guttata). Using combined immunocytochemistry for Fos and in situ hybridization for VIP, we now show that many VIP cell groups show increased transcriptional activity in response to nest building in male and female zebra finches. Particularly strong data come from the preoptic area (medial preoptic area and medial preoptic nucleus), where VIP–Fos co-expression correlates positively with three different measures of nesting behavior, as does the number of VIP-expressing cells. Remarkably, we find that VIP mRNA and/or VIP–Fos co-expression is correlated with nesting behavior in virtually every brain area that we examined, including the medial amygdala (anterior and posterior), medial bed nucleus of the stria terminalis, medial preoptic area, medial preoptic nucleus, anterior hypothalamus, ventromedial hypothalamus, periaqueductal gray complex (central gray and nucleus intercollicularis), and ventral tegmental area. Near-significant effects are also obtained in the tuberoinfundibular hypothalamus. Although most correlations are positive, negative correlations are observed for the VIP cell group of the anterior hypothalamus, a population that selectively promotes aggression, and also the periaqueductal gray complex. These data demonstrate a network-wide relationship between peptide production and social behavior that is, to our knowledge, unparalleled by other peptidergic modulators.

Status-appropriate singing behavior, testosterone and androgen receptor immunolabeling in male European starlings (Sturnus vulgaris)

Vocalizations convey information about an individual's motivational, internal, and social status. As circumstances change, individuals respond by adjusting vocal behavior accordingly. In European starlings, a male that acquires a nest site socially dominates other males and dramatically increases courtship song. Although circulating testosterone is associated with social status and vocal production it is possible that steroid receptors fine-tune status-appropriate changes in behavior. Here we explored a possible role for androgen receptors. Male starlings that acquired nest sites produced high rates of courtship song. For a subset of males this occurred even in the absence of elevated circulating testosterone. Immunolabeling for androgen receptors (ARir) was highest in the medial preoptic nucleus (POM) in males with both a nest site and elevated testosterone. For HVC, ARir was higher in dominant males with high testosterone (males that sang longer songs) than dominant males with low testosterone (males that sang shorter songs). ARir in the dorsal medial portion of the nucleus intercollicularis (DM) was elevated in males with high testosterone irrespective of dominance status. Song bout length related positively to ARir in POM, HVC and DM, and testosterone concentrations related positively to ARir in POM and DM. Results suggest that the role of testosterone in vocal behavior differs across brain regions and support the hypothesis that testosterone in POM underlies motivation, testosterone in HVC relates to song quality, and testosterone in DM stimulates vocalizations. Our data also suggest that singing may influence AR independent of testosterone and that alternative androgen-independent pathways regulate status-appropriate singing behavior.

Patterns of FOS protein induction in singing female starlings

Females of many songbird species produce song, but information about the neural correlates of singing behavior is limited in this sex. Although well studied in males, activity in premotor song control regions and social behavior regions has not been examined in females during song production. Here, we examined the immediate early gene protein product FOS in both song control and social behavior brain regions after female starlings defending nest boxes responded to an unfamiliar female in a naturalistic setting. We found that females that sang in response to the intruder had much higher numbers of fos-immunoreactive neurons (fos-ir) in the vocal control regions HVC, the robust nucleus of the arcopallium (RA), and the dorsomedial part of the nucleus intercollicularis (DM of the ICo). In HVC, fos-ir correlated positively with song length. In RA, DM and Area X, fos-ir correlated positively with number of songs produced. In social behavior regions, singers showed higher fos-ir in the nucleus taeniae of the amygdala, the dorsal part of the bed nucleus of the stria terminalis, and the ventromedial hypothalamus than non-singers. Overall, patterns of fos-ir in song control regions in females were similar to those reported for males, but differences in fos-ir were identified in social behavior regions. These differences may reflect a distinct role for brain regions involved in social behavior in female song, or they may reflect differences in the social function of female and male song.

A direct neuronal connection between the telencephalic nucleus robustus arcopallialis and the nucleus nervi hypoglossi, pars tracheosyringealis in Bengalese finches ( Lonchura striata var. domestica)

Bird species with vocal learning possess a projection from the telencephalic nucleus to the nucleus nervi hypoglossi, pars tracheosyringealis (XIIts) in the medulla, where a final common pathway that controls the vocal organ, i.e., the synrinx, originates. The anatomical basis of this projection has not been well investigated in one species of songbird, the Bengalese finch ( Lonchura striata var. domestica). The present study used anterograde and retrograde tracing experiments to examine and describe this projection in Bengalese finches. Following iontophoretic injections of biotinylated dextran amine into the telencephalic nucleus robustus arcopallialis (RA), we detected anterograde-labeled terminations in the XIIts. In addition, labeled terminals were seen in other vocal–respiratory-related nuclei, such as the dorsomedial nucleus of the nucleus intercollicularis, nucleus infraolivaris superior, nucleus of the rostral ventrolateral medulla, nucleus parambigualis, nucleus ambiguous, and nucleus retroambigualis. Furthermore, following injections into the XIIts, we detected retrograde-labeled cell bodies scattered throughout the ipsilateral RA. The present results revealed that the direct projections of the RA to the XIIts in male Bengalese finches are similar to those in other songbirds with vocal learning abilities.

Subdivisions of the auditory midbrain (n. mesencephalicus lateralis, pars dorsalis) in zebra finches using calcium-binding protein immunocytochemistry.

The midbrain nucleus mesencephalicus lateralis pars dorsalis (MLd) is thought to be the avian homologue of the central nucleus of the mammalian inferior colliculus. As such, it is a major relay in the ascending auditory pathway of all birds and in songbirds mediates the auditory feedback necessary for the learning and maintenance of song. To clarify the organization of MLd, we applied three calcium binding protein antibodies to tissue sections from the brains of adult male and female zebra finches. The staining patterns resulting from the application of parvalbumin, calbindin and calretinin antibodies differed from each other and in different parts of the nucleus. Parvalbumin-like immunoreactivity was distributed throughout the whole nucleus, as defined by the totality of the terminations of brainstem auditory afferents; in other words parvalbumin-like immunoreactivity defines the boundaries of MLd. Staining patterns of parvalbumin, calbindin and calretinin defined two regions of MLd: inner (MLd.I) and outer (MLd.O). MLd.O largely surrounds MLd.I and is distinct from the surrounding intercollicular nucleus. Unlike the case in some non-songbirds, however, the two MLd regions do not correspond to the terminal zones of the projections of the brainstem auditory nuclei angularis and laminaris, which have been found to overlap substantially throughout the nucleus in zebra finches.

Mammal-Like Organization of the Avian Midbrain Central Gray and a Reappraisal of the Intercollicular Nucleus

In mammals, rostrocaudal columns of the midbrain periaqueductal gray (PAG) regulate diverse behavioral and physiological functions, including sexual and fight-or-flight behavior, but homologous columns have not been identified in non-mammalian species. In contrast to mammals, in which the PAG lies ventral to the superior colliculus and surrounds the cerebral aqueduct, birds exhibit a hypertrophied tectum that is displaced laterally, and thus the midbrain central gray (CG) extends mediolaterally rather than dorsoventrally as in mammals. We therefore hypothesized that the avian CG is organized much like a folded open PAG. To address this hypothesis, we conducted immunohistochemical comparisons of the midbrains of mice and finches, as well as Fos studies of aggressive dominance, subordinance, non-social defense and sexual behavior in territorial and gregarious finch species. We obtained excellent support for our predictions based on the folded open model of the PAG and further showed that birds possess functional and anatomical zones that form longitudinal columns similar to those in mammals. However, distinguishing characteristics of the dorsal/dorsolateral PAG, such as a dense peptidergic innervation, a longitudinal column of neuronal nitric oxide synthase neurons, and aggression-induced Fos responses, do not lie within the classical avian CG, but in the laterally adjacent intercollicular nucleus (ICo), suggesting that much of the ICo is homologous to the dorsal PAG.

Sex Differences in the Expression of Sex Steroid Receptor mRNA in the Quail Brain

In Japanese quail, males will readily exhibit the full sequence of male-typical sexual behaviors but females never show this response, even after ovariectomy and treatment with male-typical concentrations of exogenous testosterone. Testosterone aromatisation plays a key-limiting role in the activation of this behavior but the higher aromatase activity in the brain of males compared to females is not sufficient to explain the behavioural sex difference. The cellular and molecular bases of this prominent sex difference in the functional consequences of testosterone have not been identified so far. We hypothesised that the differential expression of sex steroid receptors in specific brain areas could mediate this behavioural sex difference. Therefore, using radioactive in situ hybridisation histochemistry, we quantified the expression of the mRNA coding for the androgen receptor (AR) and the oestrogen receptors (ER) of the alpha and beta subtypes. All three receptors were expressed in an anatomically discrete manner in various nuclei of the hypothalamus and limbic system and, at usually lower densities, in a few other brain areas. In both sexes, the intensity of the hybridisation signal for all steroid receptors was highest in the medial preoptic nucleus (POM), a major site of testosterone action that is related to the activation of male sexual behaviour. Although no sex difference in the optical density of the AR hybridisation signal could be found in POM, the area covered by AR mRNA was significantly larger in males than in females, indicating a higher overall degree of AR expression in this region in males. By contrast, females tended to have significantly higher levels of AR expression than males in the lateral septum. ERalpha was more densely expressed in females than males throughout the medial preoptic and hypothalamic areas (including the POM and the medio-basal hypothalamus), an area implicated in the control of female receptivity) and in the mesencephalic nucleus intercollicularis. ERbeta was more densely expressed in the medio-basal hypothalamus of females but a difference in the reverse direction (males > females) was observed in the nucleus taeniae of the amygdala. These data suggest that a differential expression of steroid receptors in specific brain areas could mediate at least certain aspects of the sex differences in behavioural responses to testosterone, although they do not appear to be sufficient to explain the complete lack of activation by testosterone of male-typical copulatory behaviour in females.

Comparative analysis of neurogenesis between the core and shell regions of auditory areas in the chick ( Gallus gallus domesticus)

Early embryogenesis can reflect constituting organizations and evolutionary origins of brain areas. To determine whether a clear core-versus-shell distinction of neurogenesis that occurs from the auditory midbrain to the telencephalon in the reptile also appears in the bird, a single dose of [ 3H]-thymidine was injected into chick ( Gallus gallus domesticus) eggs at some successive embryonic days (E) (from E3 to E10). Towards the end of hatching, [ 3H]-thymidine labeling was examined, and the results were as follows: 1) Neuronal generation in the nucleus intercollicularis (ICo) (shell region) began at E3, whereas neurogenesis began at E4 in the nucleus mesencephalicus lateralis pars dorsalis (MLd) (core region); 2) Neurogenesis initiated at E3 in the nucleus ovoidalis (Ov) shell, but initiated at E4 in the rostral Ov core. In the medial or caudal Ov core, the percentage of heavily-labeled neurons with [ 3H]-thymidine was significantly lower at E3 age group than that in the Ov shell; 3) In field L1 and L3, two flanking regions of the primary telencephalic auditory area (field L2a), neurogenesis started at E5, but started at E6 in field L2a. These data indicate that the onset of embryogenesis began earlier in the auditory shell areas than in the core areas from the midbrain to the telencephalon. These findings provide insight into the organization of auditory nuclei and their evolution in amniotes.

Involvement of urocortinergic neurons below the midbrain central gray in the physiological response to restraint stress in pigeons

The present study was carried out to identify the diencephalic and midbrain neurons in pigeons that respond to stress (using restraint as the stressor) and determine if the urocortinergic neurons (expressing urocortin 1, Ucn1) below the midbrain central gray are among those activated. Immunolabeling for the immediate early gene Egr-1 was used to identity stress-responsive neurons, following 1–3 h of restraint. A large increase in nuclear Egr-1 immunolabeling was observed in several dorsomedial thalamic nuclei, and in a stream of neurons extending from below the mesencephalic central gray (overlapping the nucleus of Darkschewitsch at these levels) to just anterior to the nucleus of Edinger–Westphal. A more modest increase in neuronal nuclear Egr-1 was observed in the medial posterior hypothalamic area, the mesencephalic periventricular area, the ventral tegmental area, the inferior colliculus, the nucleus paramedianus of the midbrain, and the intercollicular nucleus. The distribution and abundance of urocortin-immunolabeled neurons coincided with that of the stress-responsive neurons below the mesencephalic periaqueductal gray, and about 50% of these urocortin neurons were activated by stress. These results suggest that, as in some mammals, the urocortinergic neurons of the paramedian subgriseal mesencephalon respond to stress. In those mammals, in which the boundaries of the nucleus of Edinger–Westphal are indistinct, the caudal part of the homologous field of urocortinergic neurons has been referred to as the nucleus of Edinger–Westphal. In pigeons, in which the nucleus of Edinger–Westphal is cytoarchitectonically well-defined, the caudal part of this urocortinergic field clearly does not include the nucleus of Edinger–Westphal.