Cell types of the hyperstriatum ventrale of the domestic chicken (Gallus domesticus): a Golgi study

Two major classes of neurons were observed in Golgi impregnated material of two regions of the hyperstriatum ventrale (HV) (medial and rostral HV) of the chick telencephalon: a Golgi I type neuron with long axon (putative projection neuron) and a Golgi II type neuron with short axon (putative local circuit neuron). On the basis of the nature of the axon (bifurcating or non-bifurcating, ascending or descending) and of the dendrites (spiny or aspinous, and thickness), the following subclasses (types) were identified: two long axon (projection) and three short axon (local circuit) neurons in the medial (mid-telencephalic) hyperstriatum ventrale, and one major long axon (projection) and two short axon (local circuit) neurons in the rostral hyperstriatum ventrale. Possible patterns of intraregional connections between these neurons are suggested on the basis of cytoarchitectonics. The differences in cellular types and in internal connectivity patterns strongly suggest, in agreement with physiological and behavioural observations, that the medial (mid-telencephalic) hyperstriatum ventrale distinctly differs in cellular morphology from other parts of hyperstriatum ventrale in so far as it displays a greater diversity of neuronal types with an apparent higher degree of complexity.     

Neuronal activity related to memory in the intermediate and medial part of the hyperstriatum ventrale of the chick brain

The intermediate and medial part of the hyperstriatum ventrale (IMHV) in the forebrain of the domestic chick Gallus gallus domesticus has been shown in previous studies to be critically involved in the learning process of imprinting. In the present study, 1-day-old chicks were imprinted by exposing them to one of two artificial stimuli. 24 h later each chick was given a preference test in which the two stimuli were presented in sequence. A preference score, an index of the strength of imprinting (i.e., of learning), was calculated from approach activity during the preference test. The chicks were divided into groups with low, medium and high preference scores (corresponding to weak, medium and strong learning respectively) and coded so that all subsequent procedures were performed blind. Each chick was then aneesthetized and spontaneous action potentials recorded extracellularly from groups of neurones in the left IMHV. The mean neuronal firing rate in chicks with medium and high preference scores was significantly greater than that in chicks with low preference scores. This relation between neuronal activity and preference score was not attributable to the chicks' locomotor activity per se. The results demonstrate an association between spontaneous electrical activity in the left IMHV and a measure of learning 1 day after the learning occurred.     

Termination pattern of medial hyperstriatum ventrale efferents in the archistriatum of the domestic chick

The chick archistriatum receives afferents from the intermediate part of the medial hyperstriatum ventrale (IMHV) and projects to the lobus parolfactorius (LPO). There is functional evidence to suggest that the IMHV and the LPO are connected, but there is no anatomical evidence for a direct connection between the two structures. The aim of the current study was to characterize the termination pattern of medial hyperstriatal afferents within the archistriatum to determine whether the archistriatum may act as a relay between the IMHV and LPO. Following iontophoresis of Phaseolus vulgaris leucoagglutinin into the medial hyperstriatum ventrale (including the IMHV) of 1-week-old domestic chicks, anterogradely labelled fibers were observed to descend through the medial neostriatum and paleostriatum to enter the archistriatum. These medial hyperstriatum ventrale afferents arborised profusely to give varicose axon branches within all except the anterior part of the archistriatum. However, the greatest density was present in the ventral part of the intermediate archistriatum. Electron microscope examination Phaseolus lectin immunocytochemistry and Golgi impregnation revealed that medial hyperstriatum ventrale axons formed multiple asymmetric synapses with dendritic spines (head and neck regions) on the terminal and preterminal dendritic segments of densely spiny archistriatal projection neurons. Medial hyperstriatum ventrale afferents were not observed to contact calbindin immunoreactive, presumptive “local circuit” neurons, within the archistriatum, despite a spatial overlap in their distribution. These results suggest that the archistriatum may be capable of mediating the transfer of information from the IMHV to the LPO. © 1994 Wiley-Liss, Inc.     

Differential distribution of L-aspartate- and L-glutamate-immunoreactive structures in the arcopallium and medial striatum of the domestic chick (Gallus domesticus)

The role of amino acid neurotransmitters in learning and memory is well established. We investigated the putative role of L-aspartate as a neurotransmitter in the arcopallial-medial striatal pathway, which is known to be involved in passive avoidance learning in domestic chicks. Double immunocytochemistry against L-aspartate and L-glutamate was performed at both light and electron microscopic levels. L-aspartate- and L-glutamate-immunoreactive neurons in the arcopallium and posterior amygdaloid pallium were identified and counted by using fluorescence microscopy and confocal laser scanning microscopy. Most labeled neurons of arcopallium were enriched in glutamate as well as aspartate. However, the arcopallium and posterior amygdaloid pallium differed from a neighboring telencephalic region (nidopallium; formerly neostriatum) by containing a substantial proportion of cells singly labeled for L-aspartate (15%, vs. 5.3% in the nidopallium). Aspartate-labeled neurons constitute approximately 20%, 25%, 42%, and 28% of total in the posterior amygdaloid pallium and the medial, dorsal, and anterior arcopallia, respectively. Immunoelectron microscopy showed that L-aspartate was enriched in terminals of the medial striatum. The labeled terminals had clear and round vesicles and asymmetric junctions; similar to those immunoreactive to L-glutamate. Axon terminals singly labeled for L-aspartate made up 17% of the total. In addition, 7% of neuronal perikarya and 26% of all dendritic profiles appeared to be labeled specifically with L-aspartate but not L-glutamate. The results indicate that L-aspartate may play a specific role (as distinct from that of L-glutamate) in the intrinsic and extrinsic circuits instrumental in avian learning and memory.     

Efferent connections of the dorsomedial thalamic nuclei of the domestic chick (Gallus domesticus)

Small iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin were placed in the thalamic anterior dorsomedial nucleus (DMA) of domestic chicks. The projections of the DMA covered the rostrobasal forebrain, ventral paleostriatum, nucleus accumbens, septal nuclei, Wulst, hyperstriatum ventrale, neostriatal areas, archistriatal subdivisions, dorsolateral corticoid area, numerous hypothalamic nuclei, and dorsal thalamic nuclei. The rostral DMA projects preferentially on the hypothalamus, whereas the caudal part is connected mainly to the dorsal thalamus. The DMA is also connected to the periaqueductal gray, deep tectum opticum, intercollicular nucleus, ventral tegmental area, substantia nigra, locus coeruleus, dorsal lateral mesencephalic nucleus, lateral reticular formation, nucleus papillioformis, and vestibular and cranial nerve nuclei. This pattern of connectivity is likely to reflect an important role of the avian DMA in the regulation of attention and arousal, memory formation, fear responses, affective components of pain, and hormonally mediated behaviors.     

Development of gamma-aminobutyric acid immunoreactivity in chick hyperstriatum ventrale and cerebellum: light and electron microscopical observations

The presence of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) in the avian hyperstriatum ventrale and dorsal cerebeller vermis was investigated immunocytochemically using a recently characterized antiserum raised against GABA. Tissue from domestic chicks aged from 19 days in ovo to 28 days posthatch was studied with both light and electron microscopy using pre-embedding immunocytochemistry. Basket, stellate and Golgi cells in the cerebellum, considered to be GABA-ergic, exhibited specific GABA-like immunolabelling in perikarya and in axonal and dendritic processes throughout the developmental period investigated. Purkinje cells also exhibited specific GABA-like immunoreactivity in both pre- and posthatch birds but the distribution and intensity of the immunolabelling varied with age and also in its location within the Purkinje neuron. In prehatch birds Purkinje perikarya exhibited heavy immunostaining which was substantially reduced posthatch, whereas the Purkinje primary dendrites remained immunopositive throughout the developmental period. A small population of cells in the medial hyperstriatum ventrale (mHV) were GABA-positive prehatch, but no immunopositive perikarya were evident in any posthatch samples. Small GABA-positive punctate profiles, representing boutons and transversely sectioned axons or dendrites, were present in the neuropil of all age groups studied. Possible reasons for these findings are discussed and it is suggested that the loss of perikaryal immunostaining, both in the cerebellar Purkinje cells and in those of the mHV, may be governed by maturational processes.     

Distribution of glial fibrillary acidic protein-immunopositive structures in the brain of the domestic chicken (Gallus domesticus)

The present study is the first comprehensive mapping of glial fibrillary acidic protein (GFAP)-immunopositive structures in the avian brain. Two main types of GFAP-immunopositive elements were observed: (1) nonbranching fibers, occasionally twisted or varicose, and (2) star-shaped cells. Long immunostained fibers orignate from the lateral ventricle to form three bundles. Fibers of the dorsal group, emanating from the dorsal/lateral corner of the ventricle, course in lateral, anterior, and ventral directions forming a semidome, which separates the outer pallial (lateral cortical) regions from the underlying striatal mass. The middle group of fibers is directed anteriorly and laterally corresponding to the laminae frontales superior and suprema. The ventral fiber bundle is conical and traverses the lobus parolfactorius, crossing also the lamina medullaris dorsalis (the latter consisting mainly of star-shaped cells). The hippocampus, septum, and hypothalamus also contain straight radial fibers. In some areas, given their variable orientation, the fibers cannot be regarded as merely persisting radial glia. In the telencephalon, the nuclei basalis, accumbens, ectostriatum, paleostriatum primitivum, and the ventral paleostriatum are particularly rich in GFAP-positive cells, whereas the neostriatum, hyperstriatum, and paleostriatum augmentatum are almost devoid of GFAP labelling. Certain nuclei of the thalamus and the lower brainstem are conspicuous by their low levels of GFAP immunoreactivity. The Bergmann glia were GFAP-immunonegative.     

The effects of intense sound exposure on phase locking in the chick (Gallus domesticus) cochlear nerve

Little is known about changes that occur to phase locking in the auditory nerve following exposure to intense and damaging levels of sound. The present study evaluated synchronization in the discharge patterns of cochlear nerve units collected from two groups of young chicks (Gallus domesticus), one shortly after removal from an exposure to a 120-dB, 900-Hz pure tone for 48 h and the other from a group of non-exposed control animals. Spontaneous activity, the characteristic frequency (CF), CF threshold and a phase-locked peri-stimulus time histogram were obtained for every unit in each group. Vector strength and temporal dispersion were calculated from these peri-stimulus time histograms, and plotted against the unit's CF. All parameters of unit responses were then compared between control and exposed units. The results in exposed units revealed that CF thresholds were elevated by 30-35 dB whereas spontaneous activity declined by 24%. In both control and exposed units a high degree of synchronization was observed in the low frequencies. The level of synchronization above approximately 0.5 kHz then systematically declined. The vector strengths in units recorded shortly after removal from the exposure were identical to those seen in control chicks. The deterioration in discharge activity of exposed units, seen in CF threshold and spontaneous activity, contrasted with the total absence of any overstimulation effect on synchronization. This suggested that synchronization arises from mechanisms unscathed by the acoustic trauma induced by the exposure.     

Postnatal changes in the distribution and density of neuronal nuclei and doublecortin antigens in domestic chicks (Gallus domesticus)

To understand better the rate of neurogenesis and the distribution of new neurons in posthatch domestic chicks, we describe and compare the expression of the neuronal nuclei protein (NeuN, a.k.a. Fox-3) and doublecortin antigens in the whole brain of chicks 2 days, 8 days, and 14 weeks posthatch. In the forebrain ventricular and paraventricular zones, the density of bromodeoxyuridine-, NeuN-, and doublecortin-labeled cells was compared between chicks 24 hours and 7 days after an injection of bromodeoxyuridine (2 and 8 days posthatch, respectively). The distribution of NeuN-labeled neurons was similar to Nissl-stained tissue, with the exception of some areas where neurons did not express NeuN: cerebellar Purkinje cells and olfactory bulb mitral cells. The ventral tegmental area of 2-day-old chicks was also faintly labeled. The distribution of doublecortin was similar at all timepoints, with doublecortin-labeled profiles located throughout all forebrain areas as well as in the cerebellar granule cell layer. However, doublecortin labeling was not detectable in any midbrain or brainstem areas. Our data indicate that a significant number of new neurons is still formed in the telencephalon of posthatch domestic chicks, whereas subtelencephalic areas (except for the cerebellum) finish their neuronal expansion before hatching. Most newly formed cells in chicks leave the paraventricular zone after hatching, but a pool of neurons stays in the vicinity of the ventricular zone and matures in situ within 7 days. Proliferating cells often migrate laterally along forebrain laminae into still-developing brain areas.     

Spatial correlations between the neuronal inclusions, swollen achromatic neurons, and glial cells in neuronal intermediate filament inclusion disease (NIFID)

Summary. Neuronal intermediate filament (IF) inclusion disease (NIFID) is characterized by neuronal loss, neuronal cytoplasmic IF-positive inclusions (NI), swollen neurons (SN), and a glial cell reaction. We studied the spatial correlations between the clusters of NI, SN, and glial cells in four gyri of the temporal lobe (superior temporal gyrus, inferior temporal gyrus, lateral occipitotemporal gyrus, and parahippocampal gyrus) in four cases of NIFID. The densities of histological features (per 50×250µm sample field) were as follows: NI (mean=0.41, range 0.28–0.68), SN (mean=1.41, range 0.47–2.65), glial cell nuclei (mean=5.21, range 3.63–8.17). The NI and the SN were positively correlated in half of the brain regions examined, the correlations being present at the smallest field size (50×250µm). The NI were also positively or negatively correlated with the glial cell nuclei in different areas, the negative correlations being present at the smallest field size. Glial cell nuclei were positively or negatively correlated with the SN in different brain areas, mainly at the larger field sizes (400×250 and 800×250µm). The spatial correlation between the clusters of NI and SN in the cortex suggests their development within the same columns of cells. At first, the glial cell reaction is also confined to these columns but later becomes more generally distributed across the cortex.     

Afferent connections of septal nuclei of the domestic chick (Gallus domesticus): a retrograde pathway tracing study

The afferents to the septum of the domestic chicken were studied using retrograde tracers, rhodamine conjugated latex bead or Fast Blue, placed in different septal subregions. The results were verified by anterograde tracer injections deposited to selected areas. The main telencephalic afferents to the septum arise ipsilaterally from the hippocampal formation, dorsolateral corticoid area, piriform cortex, amygdaloid pallium, and the ventral pallidum. Contralateral afferents originate from the lateral septum and the amygdaloid pallium. A massive bilateral projection arises from the lateral hypothalamus. Other hypothalamic afferents arise from the periventricular, paraventricular and anterior medial nuclei, and the premammillary and mammillary areas. The dorsal thalamic nuclei (dorsal medial anterior and posterior) and the reticular dorsal nuclei also contribute septal afferents. Brainstem afferents arise bilaterally from the ventral tegmental area, substantia nigra, central gray, A8, locus coeruleus, ventral subcoeruleus nucleus, and raphe nuclei. The main terminal fields for septal afferents lie in the lateral septal nucleus and the belt of medial septal nucleus. The core of the latter is invaded mainly by fibers from the brainstem, presumably belonging to the ascending activating system. The septal afferents of the chicken are largely similar to those of other avian and nonavian species. The most prominent differences with previous pigeon data were found in the subregional selectivity of the hippocampal formation, dorsolateral corticoid area, mammillary nuclei, some dorsal thalamic nuclei, substantia nigra, and subcoeruleus nuclei in their projections to defined septal nuclei.     

Neural structures associated with reproductive development and photoperiodic effects in male chicks (Gallus domesticus)

White Leghorn chicks given parasagittal knife cuts that isolated the hypothalamus from lateral connections displayed early maturation of the testes. When the operated chicks were subsequently exposed to a long-day photoperiod known to stimulate gonadal development, no additional increase in the size of the testes occurred. In contrast, controls demonstrated a marked development of the gonads that surpassed testes size of experimentals. An additional study was designed to determine whether the archistriatum, a telencephalic structure that has significant projections to the hypothalamus, may be involved in the earlier onset of puberty shown by knife-cut chicks. Three different sets of brain cuts were made that severed projections from the archistriatum to the hypothalamus. None of the knife cuts affected development of the gonads when the birds were examined six weeks after surgery. Two of the three sets of knife cuts, however, effected a significant reduction in food intake, and operated chicks also displayed a significantly lower body weight compared to controls (p less than 0.05).     

Distinct mechanisms for expression of Fos-like immunoreactivity and synaptic potentiation in telencephalic hyperstriatum of the quail chick

In the intermediate and medial hyperstriatum ventrale (IMHV), a telencephalic region essentially involved in the initial processes of early learning tasks in poultry chicks, induction of an immediate early gene c-fos correlates significantly with the degree of learning (K.V. Anokhin, R. Mileusnic, I.Y. Shamakina, S.P.R. Rose, Effects of early experience on c-fos gene expression in the chick forebrain, Brain Res. 544 (1991) 101–107; B.J. McCabe, G. Horn, Learning-related changes in Fos-like immunoreactivity in the chick forebrain after imprinting, Proc. Natl. Acad. Sci. USA 91 (1994) 11417–11421). In slices of IMHV in vitro, on the other hand, tetanic stimulation at a low frequency induces a potentiation of synaptic responses (P.M. Bradley, B.D. Burns, A.C. Webb, Potentiation of synaptic responses in slices from the chick forebrain, Proc. R. Soc. Lond. B. 243 (1991) 19–24; T. Matsushima, K. Aoki, Potentiation and depotentiation of DNQX-sensitive fast excitatory synaptic transmission in telencephalon of the quail chick, Neurosci. Lett. 185 (1995) 179–182). In this study, we have examined a possible causal link between these two forms of activity-dependent processes, c-fos expression and synaptic potentiation. C-fos was visualized immunohistochemically using antibody raised against the Fos-protein, and potentiation was evaluated on the basis of field potential responses to local electrical stimulation. Tetanic stimulation (5 Hz×300 pulses) was required for potentiation, but not for c-fos expression. Conversely, a negative correlation appeared between them, and slices with relatively high density of Fos-like immunoreactive cells around the stimulation site failed to show potentiation. Furthermore, drugs similarly effective in blocking potentiation (such as AP5 (NMDA receptor antagonist) and bicuculline (GABA_A receptor antagonist)) had different effects on the c-fos induction. While AP5 had minor, if any, effects on c-fos expression, bicuculline enhanced it selectively around the site of stimulation. Our results suggest that these two processes are basically distinct, and could represent different aspects in the formation of memory traces in IMHV.     

3H]naloxone binding in the brain of the domestic chick (Gallus domesticus) determined by in vitro quantitative autoradiography

The distribution of [3H]naloxone binding sites was investigated in the brains of one-day-old domestic chicks using quantitative receptor autoradiography. Among the forebrain regions, the greatest levels of opiate binding were found in hyperstriatum dorsale, hyperstriatum ventrale, hyperstriatum intercalatum supremum and neostriatum. Intermediate levels were found throughout the paleostriatal regions, septum, thalamus, archistriatum, hyperstriatum accessorium and area parahippocampalis whilst in hippocampus and ectostriatum the density of [3H]naloxone binding sites was low. In the hindbrain, high levels of opiate binding were found in optic tectum whereas in cerebellum the density of binding sites was barely above background. The greatest densities of opiate binding appear to coincide with regions found to be involved in sensory processing and memory storage.     

Antigenic compartmentation of the cerebellar cortex in the chicken (Gallus domesticus)

The chick is a well‐understood developmental model of cerebellar pattern formation,but we know much less about the patterning of the adult chicken cerebellum. Therefore an expression study of two Purkinje cell stripe antigens—zebrin II/aldolase C and phospholipase Cβ4 (PLCβ4)—has been carried out in the adult chicken (Gallus domesticus). The mammalian cerebellar cortex is built around transverse expression domains (“transverse zones”), each of which is further subdivided into parasagittally oriented stripes. The results from the adult chicken reveal a similar pattern. Five distinct transverse domains were identified. In the anterior lobe a uniformly zebrin II‐immunopositive/PLCβ4‐immunonegative lingular zone (LZ; lobule I) and a striped anterior zone (AZ; lobules II–VIa) were distinguished. A central zone (CZ; ∼lobules VIa–VIIIa,b) and a posterior zone (PZ; ∼lobules VIIIa,b–IXc,d) were distinguished in the posterior lobe. Finally, the nodular zone (NZ; lobule X) is uniformly zebrin II‐immunoreactive and is innervated by vestibular mossy fibers. Lobule IXc,d is considered as a transitional region between the PZ and the NZ, because the vestibular mossy fiber projection extends into these lobules and because they receive optokinetic mossy and climbing fiber input. It is proposed that the zebrin II‐immunonegative P3‐ stripe corresponds to the lateral vermal B zone of the mammalian cerebellum and that the border between the avian homologs of the mammalian vermis and hemispheres is located immediately lateral to P3−. Thus, there seem to be transverse zones in chicken that are plausible homologs of those identified in mammals, together with an LZ that is characteristic of birds. J. Comp. Neurol. 518:2221–2239, 2010. © 2010 Wiley‐Liss, Inc.     

The organisation of the basal ganglia in the domestic chick (Gallus domesticus): anatomical localisation of DARPP-32 in relation to glutamate

Subpallial structures are highly conserved across the different vertebrate species. They are instrumental in the neural processing relevant to adaptive learning, decision making, motivation and behavioural strategies. Of the striatal regions, our attention has been focussed on the medial and ventral striatum (MSt), now parcellated into subregions, and also including the nucleus accumbens (Ac). Similar to mammals, the avian Ac and MSt receive glutamatergic input from the pallium and dopaminergic input from the substantia nigra and ventral tegmental area. Coincidence between glutamatergic and dopaminergic synaptic activities in the ventral/medial striatum, including the Ac, is required for memory to be formed for a given pairing of stimulus and a hedonic quality or behavioural salience. The underlying mechanism involves the activation of NMDA and dopaminergic receptors, as well as the phosphorylation of dopamine-cAMP-regulated phosphoprotein (DARPP-32). Using quantitative electron microscopy of chick specimens double-labelled against glutamate and DARPP-32 we observed direct synaptic connections between glutamate immunoreactive axon terminals and DARPP-32 labelled dendrites in the MSt and also in the posterolateral telencephalon (nidopallium caudolaterale, a prefrontal cortex equivalent region) and the hippocampus. Glutamate immunoreactive axons synapsed with both DARPP-32 immunoreactive (DARPP-32+) and DARPP-32 negative (DARPP-32−) dendrites, forming asymmetrical junctions, in all brain regions observed. The existence of direct synaptic contacts between excitatory amino acid containing axon terminals and DARPP-32 containing dopaminoceptive neurons of the chicken MSt underlines the functional homology with mammalian striatal systems.     

Changes in the number and structure of dendritic spines 25 hours after passive avoidance training in the domestic chick, Gallus domesticus

One-day-old chicks spontaneously peck at a shiny chrome bead. If the bead is coated with methylanthranilate, a bitter tasting substance (M-chicks), they peck once and avoid a similar bead subsequently. Control chicks peck, and continue pecking at a bead dipped in water (W-chicks). Twenty-five hours after this one-trial passive avoidance training the brains were fixed and Golgi-impregnated. A class of large, multipolar, projection neurons from the intermediate medial hyperstriatum ventrale (IMHV) of both hemispheres from M- and W-chicks were examined for changes in their spine density and spine shape. An estimate of the true spine number was obtained using the correction formula of Feldman and Peters. M-chicks showed highly significant increases (P less than 0.0001) in spine density of between 89-113% in the left hemisphere, and 37-69% in the right, compared with W-chicks. There was a significant hemispheric asymmetry in W-chicks: the right hemisphere had approximately 47% more spines per micron than the left, and this difference was abolished after training. Following passive avoidance training, significant increases in spine head diameter (by approximately 9%) and decreases in spine stem length (by approximately 17%), with no significant alterations in overall spine length, were observed in the left hemisphere. The mean dendrite lengths were not significantly changed after training, but an asymmetry of this measure in W-chicks (left greater than right) was present in dendrite orders 2 (P less than 0.01) and 3 (P less than 0.02). These results show that spine densities can increase rapidly (within 25 h) following a one-trial passive avoidance training task and that spine shape changes can be found on the same dendrites which also show changes in spine number. The data support the view that dendritic spines are involved in memory formation processes.