Pigeon Brain Research Articles

Molecular Cloning and Differential Expression Patterns of Avian Glutamate Receptor mRNAs

We have identified and studied potential ionotropic glutamate receptor genes in pigeon brain. Three cDNA clones exhibit significant amino acid sequence identity to members of a rodent ligand-gated ion channel family. One of them, GluP-II, encodes a full-length AMPA-sensitive glutamate receptor GluR2 (GluR-B) homologue, whereas the other two partial clones, designated as GluP-III and -IV, are nearly identical to rodent GluR3 (GluR-C) and GluR4 (GluR-D) receptor subunits. Northern analysis demonstrated that the avian genes are widely expressed in the brain. Within the brain regions analyzed by in situ hybridization histochemistry, the three avian GluR subunits showed distinct and regionally specific mRNA expression patterns in the adult. Most of the differences in their expression were observed in cell types of the telencephalon, certain thalamic nuclei, the optic tectum, and the cerebellar cortex. A particularly striking finding was the expression of GluP-II in Golgi epithelial/Bergmann glial cells. In contrast, Bergmann glial cells in rat cerebellum do not express GluR2 (GluR-B) subunit genes. Immunoreactivity for a monoclonal sequence-specific antipeptide antibody was widespread and most prominent in Purkinje cell perikarya and their dendrites, neuronal cell bodies of the ectostriatum, and the deep optic tectum. These results demonstrate the existence of multiple subunits of the ionotropic glutamate receptor channel family in avians. Excitatory amino acid receptor genes appear to be highly conserved during evolution.

Kinetics of the inhibition of acetylcholinesterase from pigeon brain by procaine hydrochloride

The kinetic parameters of the inhibition of pigeon brain acetylchlolinesterase (AChE) by procaine hydrochloride were investigated. Procaine (0·083–1·67 mM) reversibly inhibited AChE activity (15–83 percent) in a concentration dependent manner, the IC50 being about 0·38 mM. The Michaelis-Menten constant (Km) for the hydrolysis of acetylthiocholine iodide was found to be 1·53 × 10−4 M and the Vmax was 1·06 μmol min−1 mg−1 protein. Dixon as well as Lineweaver-Burk plots and their secondary replots indicated that the nature of the inhibition is of the linear mixed type which is considered to be a mixture of partial competitive and pure non-competitive. The values of Ki(slope) and Ki (intercepts) were estimated as 0·14 mM and 0·22 mM respectively by the primary Dixon and by the secondary replots of the Lineweaver-Burk plot. The Ki′/Ki ratio shows that procaine has a greater affinity of binding for the peripheral than for the active site.

Pharmacologic characterization of melatonin-mediated phosphoinositide hydrolysis in pigeon brain.

High densities of [125I]-iodomelatonin binding sites have been demonstrated in pigeon brain. Melatonin binding sites have been shown to be linked to signal transduction mechanisms in other species. The present study investigated the melatonin-mediated second messenger response of phosphoinositide hydrolysis in slices of telencephalon, optic tectum, cerebellum, hypothalamus, and pons medulla of pigeon brain. The highest rates of melatonin-mediated phosphoinositide hydrolysis were observed in telencephalon and pons/medulla. Relative potencies of melatonin agonists to induce phosphoinositide hydrolysis were as follows: 2-iodomelatonin > 6-chloromelatonin > N-acetylserotonin > melatonin > > serotonin (5-HT). Agonist-induced phosphoinositide hydrolysis was blocked by N-acetyltryptamine (NAT), a melatonin antagonist, but not by ketanserin, a 5HT2A/2C receptor antagonist, demonstrating that phosphoinositide hydrolysis did not result from 5HT2A or 5HT2C receptor stimulation. In addition, the effects of melatonin agonists were sensitive to prazosin, an alpha-adrenergic antagonist reported to exhibit nanomolar affinity for melatonin binding sites in hamster brain, but not to phentolamine, an alpha-adrenergic antagonist that shows no affinity for melatonin binding sites. These data provide evidence that signal transduction associated with melatonin in pigeon brain involves the induction of phosphoinositide hydrolysis as a second messenger.

Distribution of choline acetyltransferase immunoreactivity in the pigeon brain

We have investigated the distribution of cholinergic perikarya and fibers in the brain of the pigeon (Columba livia). With this aim, pigeon brain sections were processed immunohistochemically by using an antiserum specific for chicken choline acetyltransferase. Our results show cholinergic neurons in the pigeon basal telencephalon, the hypothalamus, the habenula, the pretectum, the midbrain tectum, the dorsal isthmus,the isthmic tegmentum, and the cranial nerve motor nuclei. Cholinergic fibers were prominent in the dorsal telencephalon, the striatum, the thalamus, the tectum, and the interpeduncular nucleus. Comparison of our results with previous studies in birds suggests some major cholinergic pathways in the avian brain and clarifies the possible origin of the cholinergic innervation of some parts of the avian brain. In addition, comparison of our results in birds with those in other vertebrate species shows that the organization of the cholinergic systems in many regions of the avian brain (such as the basal forebrain, the epithalamus, the isthmus, and the hindbrain) is much like that in reptiles and mammals. In contrast, however, birds appear largely to lack intrinsic cholinergic neurons in the dorsal ("neocortex-like") parts of the telencephalon.

Expression and Localization of Tissue Factor-Based Procoagulant Activity (PCA) in Pigeon Monocyte-Derived Macrophages

Monocyte-derived macrophages, focal to initiation and progression of atherosclerosis, have been implicated in thrombotic complication of this disease. In the present study we demonstrated tissue factor based procoagulant activity in cultured macrophages from the White Carneau pigeon following endotoxin (1-2 micrograms/ml) stimulation. This macrophage procoagulant activity paralleled activity obtained with pigeon brain homogenate. We used Enzyme-Linked Coagulation Assay (ELCA), an ultrasensitive microtiter plate assay, to measure procoagulant activity in these cells. Through the use of clotting factors purified from pigeon plasma, procoagulant activity could be detected with as few as 1-3 cells. Tissue factor antigen, detected through the use of immunogold labelling in conjunction with a polyclonal antibody which was highly specific to human tissue factor, was distributed uniformly over the plasma membrane of the endotoxin-stimulated cells. These studies suggest that this procoagulant activity might play an important role in the pathobiology of atherosclerosis in White Carneau pigeons by initiating fibrin polymerization and thus leading to thrombotic complications of the disease.

Distribution of neuropeptide Y, substance P, and choline acetyltransferase in the developing visual system of the pigeon and effects of unilateral retina removal.

The distribution of three neuroactive substances, neuropeptide Y, substance P, and choline acetyltransferase, was studied by immunocytochemical methods in central visual regions of adult, developing, and ablated pigeon brains. In normal adult brains, neuropeptide Y-positive cells and processes were present in the nucleus pretectalis, the nucleus of the basal optic root, the nucleus of the marginal optic tract, and the visual Wulst. Substance P-positive cells and processes were found in the optic tectum and in the visual Wulst. Stained fibers and terminal-like processes, but no cells, were also observed in several visual thalamic nuclei. Choline acetyltransferase-positive cells and processes were located in the optic tectum, visual Wulst, the nucleus isthmo opticus, nucleus isthmi and certain visual thalamic nuclei. Cholinergic fibers and processes, but no cells, were present in the nucleus principalis precommissuralis, the supraoptic decussation, and the nucleus lentiformis mesencephali, pars magnocellularis. In the course of development, the distribution of immunoreactivity for all three substances was found to vary. These changes often involved either progressive increases or decreases in the density of labeled cells, neuropil and/or terminal-like profiles. Experiments with retina ablated pigeons clearly demonstrated that changes in the normal pattern of immunoreactivity distribution only occurred if the retina was removed immediately after hatching, i.e., before retinofugal connections have been established. The adult pattern of immunoreactivity for all three substances appears to be reached at about the same time that the anatomical and functional maturation of the pigeon visual system is completed. The present results suggest that this temporal correlation reflects the important role that retinal afferents play in the development of these putative peptidergic and cholinergic systems.

Time to collision is signalled by neurons in the nucleus rotundus of pigeons.

Throughout the animal kingdom, the sight of a rapidly approaching object usually signals danger and elicits an escape response. Gibson suggested that the symmetrical expansion of an object's image (looming) is the critical variable determining that the object is on a collision course with the observer. Similarly, large expanding flow-fields like those produced by locomotion may precipitate manoeuvres such as turning or landing. From such observations it has been shown that the optic flow parameter, tau, which specifies time to contact with the approaching object best fits the behavioural data. We describe a subpopulation of neurons in the nucleus rotundus of the pigeon brain that respond selectively to objects moving on a collision course towards the bird.

Vasoactive intestinal peptide binding sites and fibers in the brain of the pigeon Columba livia: An autoradiographic and immunohistochemical study

The distribution of vasoactive intestinal peptide (VIP) binding sites in the pigeon brain was examined by in vitro autoradiography on slide-mounted sections. A fully characterized monoiodinated form of VIP, which maintains the biological activity of the native peptide, was used throughout this study. The highest densities of binding sites were observed in the hyperstriatum dorsale, archistriatum, auditory field L of neostriatum, area corticoidea dorsolateralis and temporo-parieto-occipitalis, area parahippocampalis, tectum opticum, nucleus dorsomedialis anterior thalami, and in the periventricular area of the hypothalamus. Lower densities of specific binding occurred in the neostriatum, hyperstriatum ventrale and nucleus septi lateralis, dorsolateral area of the thalamus, and lateral and posteromedial hypothalamus. Very low to background levels of VIP binding were detected in the ectostriatum, paleostriatum primitivum, paleostriatum augmentatum, lobus parolfactorius, nucleus accumbens, most of the brainstem, and the cerebellum. The distribution of VIP-containing fibers and terminals was examined by indirect immunofluorescence using a polyclonal antibody against porcine VIP. Fibers and terminals were observed in the area corticoidea dorsolateralis, area parahippocampalis, hippocampus, hyperstriatum accessorium, hyperstriatum dorsale, archistriatum, tuberculum olfactorium, nuclei dorsolateralis and dorsomedialis of the thalamus, and throughout the hypothalamus and the median eminence. Long projecting fibers were visualized in the tractus septohippocampalis. In the brainstem VIP immunoreactive fibers and terminals were observed mainly in the substantia grisea centralis, fasciculus longitudinalis medialis, lemniscus lateralis, and in the area surrounding the nuclei of the 7th, 9th, and 10th cranial nerves. The correlation between the distribution of VIP binding sites and immunoreactive fibers and terminals was assessed in a restricted number of regions. A qualitatively good matching was found in the area corticoidea dorsolateralis, hyperstriatum dorsale, hyperstriatum accessorium, nucleus septi lateralis, nuclei dorsomedialis and dorsolateralis thalami, and in some hypothalamic areas. A striking mismatch occurred in the hyperstriatum ventrale, neostriatum, tectum opticum (high to moderate density of binding sites but only few immunoreactive profiles), and in the tuberculum olfactorium, median eminence, and spinal cord (lower density of binding sites but abundant immunoreactive profiles). The paleostriatum, lobus parolfactorius, and ectostriatum were virtually devoid of both binding sites and immunoreactive profiles. The results are discussed in relation to the known actions of VIP in the rodent and avian brain and are compared with previous observations on the distribution of VIP binding sites in the central nervous system of other vertebrates.

125I]Iodomelatonin-binding sites in the pigeon brain: binding characteristics, regional distribution and diurnal variation

The binding and pharmacological characteristics of melatonin-binding sites labelled by [125I]iodomelatonin in membrane preparations from the pigeon brain were determined. Specific binding of [125I]iodomelatonin in the membrane preparations of pigeon brain was rapid, stable, saturable and reversible. The [125I]iodomelatonin-binding sites had the following order of pharmacological affinities: melatonin greater than 6-chloromelatonin greater than N-acetylserotonin much greater than 5-hydroxytryptamine greater than tryptamine greater than 5-methoxytryptophol, greater than 1-acetylindole-3-carboxytryptamine, 5-hydroxyindole-3-acetic acid, L-tryptophan and 3-acetylindole. Compounds known to act on serotonin receptors, adrenoceptors or cholinoceptors were inactive compared with melatonin. Of the various brain regions studied, melatonin binding was greatest in the hypothalamus, intermediate in the mid-brain, pons-medulla and telencephalon, and low in the cerebellum. Subcellular fraction studies indicated that 39% of the binding was located in the mitochondrial fraction, 34% in the nuclear fraction, 21% in the microsomal fraction and 5.6% in the cytosol fraction. Scatchard analysis of the membrane preparations revealed a dissociation constant (Kd) of 206.3 +/- 57.9 pmol/l and a total number of binding sites (Bmax) of 26.7 +/- 1.9 fmol/mg protein in the middle of the light period (mid-light). In addition, saturation studies demonstrated that [125I]iodomelatonin-binding sites in pigeon brain membrane preparations were 36.2% higher at mid-light (26.7 +/- 1.9 fmol/mg protein) than in the middle of the dark period (19.6 +/- 1.25 fmol/mg protein), with no significant variation in their binding affinities.

Chromatin Higher-Order Structure: Two-Start Double Superhelix Formed by Zig-Zag Shaped Nucleosome Chain with Folded Linker DNA

Hydrodynamic properties of chromatins differing in linker DNA length and in transcriptional activity have been studied by the method of sedimentation velocity. Oligonucleosomes of different chain length were isolated from chromatins of pigeon brain cortical neurones, rat thymus and sea urchin sperm characterized by nucleosome DNA repeat length of 165, 198 and 248 base pairs respectively. The hydrodynamic behaviour of oligonucleosomes in the dependence on the number of nucleosomes in the chain and on the ionic strength has been analysed on the basis of cylinder model. The data obtained allows one to calculate the main structural parameters of the oligonucleosomal chain: its mass per unit length, the hydrodynamic diameter of the chain, the length of the chain per nucleosome and DNA packing ratio. It is shown that hydrodynamic behaviour of nucleosome oligomers from all types of chromatins investigated at low ionic strength can be well described by the model of three-dimensional zig-zag chain with similar diameter and length of the chain per nucleosome, DNA packing ratio growing with the increase of linker DNA length. It can be achieved by unfolding the short linker DNA in neurone chromatin and by coiling the long linker DNA of sea urchin sperm chromatin into a loop. With the increase of ionic strength zig-zag shaped nucleosomal chain is condensed into a two-start double superhelix with closely arranged nucleosomes and linker DNA loops packed inside the superhelix. The suggested model is in good agreement with available experimental data and overcomes a number of difficulties which arise for the solenoid model and other models of the 30-nm chromatin fibril.

Autoradiographic analysis of the distribution of vasoactive intestinal peptide binding sites in the vertebrate central nervous system: a phylogenetic study

The distribution of vasoactive intestinal peptide (VIP) binding sites in the brain of several vertebrate species was examined by in vitro autoradiography on slide-mounted sections. This study included fish, frog, snake, pigeon, rat, mouse, guinea pig, cat and monkey brain. A fully characterized, monoiodinated form of vasoactive intestinal peptide (M- 125I-VIP), which maintains the biological activity of the native peptide in the central nervous system (CNS), was used throughout the study. Among the lower vertebrate species, no significant specific binding was found in the fish brain, whereas in the frog and snake brain, specific VIP binding sites were observed, mainly in the telencephalon. In the pigeon brain, high densities of VIP binding sites were localized in the hyperstriatum, neostriatum, archistriatum, hippocampal area, dorsolateral cortical area and in the optic tectum. Ectostriatum and paleostriatum augmentatum displayed lower densities of specific binding. In mammals, the highest concentrations of VIP binding sites were observed in the rodent brain. In the rat, mouse and guinea pig brain, high densities were detected in the olfactory bulb, external layers of the cerebral cortex, dentate gyrus, midline thalamic nuclei, geniculate nuclei, some hypothalamic nuclei, superior colliculus coeruleus. Intermediate densities were found in amygdala, caudate-putamen, septum and nucleus accumbens, CA1–CA3 fields of the hippocampus and central gray. The cerebellum of these species presented high densities of VIP binding sites, with species to species differences in their localization. The non-specific binding was, however, increased in the rodent cerebellum. Lower densities of VIP binding sites were observed in the cat and monkey CNS. In these two species, the non-specific binding was considerably higher than in the lower mammals brain. In the cat and monkey brain, as in the lower mammals, the highest densities were revealed in the neocortex, dentate gyrus, thalamic nuclei and some midbrain structures including substantia nigra and locus coeruleus. In all the species studied, the white matter was never labeled with M- 125I-VIP. This study suggests that VIP binding sites appear relatively early in the evolution of the vertebrate CNS. The most important densities of specific VIP binding sites are observed in the pigeon and rodent brain, whereas the cat and monkey present a marked increase in non-specific binding. It is interesting to note that the distribution of VIP binding sites as revealed by autoradiography is quite conservative in terms of evolution and indicates an association, although non-exclusive, of VIP receptors with brain regions involved in the processing of specific sensory inputs.

5.HT1 receptors in the vertebrate brain

The regional distribution of high affinity [3H]5-HT recognition sites in the brain of several vertebrates (pigeon, rat, mouse, guinea-pig, cat, dog, monkey and human) was analyzed using in vitro autoradiography. The presence of subtypes of 5-HT1 binding sites was investigated by selective displacements with 8-OH-DPAT, mesulergine and (+/-)SDZ 21-009 at appropriate concentrations to block 5-HT1A, 5-HT1C and 5-HT1B sites respectively. In addition, 5-HT1A and 5-HT1C sites were directly visualized with the more selective radioligands [3H]8-OH-DPAT and [3H]mesulergine, respectively. In the pigeon brain, total [3H]5-HT binding sites were enriched in all telencephalic areas. Densely labelled regions were also present in the optic tectum and the brainstem. No binding was observed in the cerebellum. 8-OH-DPAT and mesulergine only displaced a small proportion of [3H]5-HT binding in most of the areas where high concentrations of 5-HT1 sites were found. (+/-)SDZ 21-009 did not affect [3H]5-HT binding in the regions examined. Taking into account our pharmacological studies, these results suggest that the majority of 5-HT1 sites belong to the 5-HT1D subtype in the pigeon brain. In the mammalian species investigated high levels of [3H]5-HT binding were found in the neo-cortex, hippocampal formation, basal ganglia and related structures (substantia nigra), raphe dorsalis, nucleus superior colliculus and choroid plexus. However, these brain areas were differentially enriched in subtypes of 5-HT1 recognition sites.(ABSTRACT TRUNCATED AT 250 WORDS)

5-HT1D receptors in guinea-pig and pigeon brain

The characteristics of high affinity [3H]5-HT (5-hydroxytryptamine) binding to non 5-HT1A non 5-HT1C sites were examined in crude membranes prepared from different regions of guinea-pig and pigeon brains. The coupling of these sites to adenylate cyclase was examined, and its pharmacological profile investigated. In the presence of 100 nmol/l 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) and 100 nmol/l mesulergine, [3H]5-HT labelled with nanomolar affinity an apparently homogeneous population of recognition sites in guinea-pig and pigeon brain membranes. The rank order of affinities of agonists and antagonists (5-CT (5-carboxamidotryptamine) greater than 5-HT greater than RU 24969 (5-methoxy-3-(1,2,3,6-tetrahydro-4- pyridinyl)-1H indole succinate) greater than yohimbine greater than or equal to rauwolscine greater than DP-5-CT (N,N dipropyl-5-carboxamidotryptamine) greater than or equal to mianserin greater than 8-OH-DPAT greater than mesulergine greater than SDZ 21-009 ((+/-)-4(3-tert-butyl-amino-2-hydroxypropoxy)-indol-2 carbonic acid isopropyl ester) greater than (-)propranolol), as well as their individual pKD values, were very similar to those at porcine caudate 5-HT1D sites and clearly different from those at rat cortex 5-HT1B sites. In the substantia nigra of the guinea-pig the 5-HT receptor-mediated inhibition of forskolin-stimulated adenylate cyclase had a pharmacological profile fully comparable to that of 5-HT1D binding sites (5-CT greater than 5-HT greater than yohimbine greater than RU 24969 greater than 8-OH-DPAT greater than SDZ 21-009 = isamoltane greater than (-)pindolol greater than (-)propranolol).(ABSTRACT TRUNCATED AT 250 WORDS)

CNS thermosensitivity and control of latent heat dissipation in the pigeon

Thermoregulatory responses to heat exposure were studied in 12 hand-reared, acclimated pigeons (Columbia livia). Measurements of body temperature (Tcl), brain temperature (Tbr), cutaneous water evaporation (CWE) and respiratory frequency (fr) were carried out in intact conscious heat exposed birds. In a second group of lightly restrained birds, fr and CWE were taken when temperatures of the trunk, brain and air (Ta) were independently changed. Increasing Tbr to 43.5–43.8°C induced a pronounced polypnea (deep and fast, (300 breaths min −1) when Tcl regulated at 42.4°C. Moreover, when hyperthermia (Tcl = 43.0° C) was combined with increased Tbr (43.0–43.8°C) shallow and fast panting (>500 breaths min −1) was evoked. CWE was probably elicited by inputs generated by the skin warm receptors as a result of increased Ta. Moreover it was demonstrated that warming the brain to 42.5°C elicits cutaneous water evaporation in birds exposed to 26°C. When a high Ta (60°C) is accompanied by a high relative humidity (17%), the combined effect generates inputs eliciting intensive panting. The integration of the present and earlier data allows us to generate a model demonstrating the distinguished significance of the trunk, skin and brain thermosensors in the regulation of both respiratory and cutaneous latent heat dissipation. The present model also emphasizes the fact that the highly thermosensitive pigeon brain responds in a similar pattern to that found in mammals

Distribution of mu, delta, and kappa opiate receptor types in the forebrain and midbrain of pigeons

Ligands that are highly specific for the mu, delta, and kappa opiate receptor binding sites in mammalian brains have been identified and used to map the distribution of these receptor types in the brains of various mammalian species. In the present study, the selectivity and binding characteristics in the pigeon brain of three such ligands were examined by in vitro receptor binding techniques and found to be similar to those reported in previous studies on mammalian species. These ligands were then used in conjunction with autoradiographic receptor binding techniques to study the distribution of mu, delta, and kappa opiate receptor binding sites in the forebrain and midbrain of pigeons. The autoradiographic results indicated that the three opiate receptor types showed similar but not identical distributions. For example, mu, delta, and kappa receptors were all abundant within several parts of the cortical-equivalent region of the telencephalon, particularly the hyperstriatum ventrale and the medial neostriatum. In contrast, in other parts of the cortical-equivalent region of the avian telencephalon, such as the dorsal archistriatum and caudal neostriatum, only kappa receptors appeared to be abundant. Within the basal ganglia, all three types of opiate receptors were abundant in the striatum and low in the pallidum. Within the diencephalon, kappa and delta binding was high in the dorsal and dorsomedial thalamic nuclei, but the levels of all three receptor types were generally low in the specific sensory relay nuclei of the thalamus. Kappa binding and delta binding were high, but mu was low in the hypothalamus. Within the midbrain, all three receptor types were abundant in both the superficial and deep tectal layers, in periventricular areas, and in the tegmental dopaminergic cell groups. In many cases, the distribution of opiate receptors in the pigeon forebrain generally showed considerable overlap with the distribution of opioid peptide-containing fiber systems (for example, in the striatal portion of the basal ganglia), but there were some clear examples of receptor-ligand mismatch. For example, although all three receptor types are very abundant in the hyperstriatum ventrale, opioid peptide-containing fibers are sparse in this region. Conversely, within the pallidal portion of the basal ganglia, opioid peptide-containing fibers are abundant, but the levels of opiate receptors appear to be considerably lower than would be expected. Thus, receptor-ligand mismatches are not restricted to the mammalian brain, since they are a prominent feature of the organization of the brain opiate systems in pigeons.(ABSTRACT TRUNCATED AT 400 WORDS)

Topographical distribution of CRF immunoreactivity in the pigeon brain

The distribution of corticotropin-releasing factor (CRF) immunoreactivity was demonstrated by immunocytochemistry in intact and colchicine-treated pigeons. Colchicine injections were administered at different times related to the circadian activity of the CRF-adrenocorticotropin (ACTH)-corticosterone axis. Three CRF antisera were used, two directed against synthetic rat CRF and one directed against synthetic ovine CRF. No fundamental differences appeared in the pigeon brain with respect to the specific CRF antiserum used. The most effective colchicine injection times corresponded to hypersecretion in the corticotropic axis. CRF-immunopositive neurons were scattered throughout the pigeon brain. In addition to the paraventricular hypothalamic system, which is involved in adenohypophysial ACTH regulation, several other hypothalamic and extrahypothalamic areas showed CRF neurons. The distribution suggests that CRF may also act as a modulator and a neurotransmitter. Two hypothalamic paraventricular nucleus-median eminence CRF pathways are described here. Moreover, CRF-immunopositive reactions were observed in specific areas of cerebral ventricle walls, suggesting that CRF may be released into the cerebral fluid.

Distribution of GABA-like immunoreactivity in the pigeon brain

The distribution of GABA-like immunoreactivity in glutaraldehyde-fixed pigeon brains was studied by means of a monoclonal antibody. GABA-like immunoreactivity was observed in neuronal perikarya of different sizes as well as in neuropil and in certain fiber tracts. Certain staining patterns indicated the existence of several GABAergic projection systems in the pigeon brain. Indeed, a high density of immunostained perikarya and a low density of labeled terminal-like elements was the prominent pattern in the nuclei subpretectalis and posteroventralis, while an absence of perikaryal GABA-like immunoreactivity and accumulations of immunoreactive dots were observed in the isthmo-optic nucleus, amongst others. In the optic tectum, stained cell bodies with radially oriented processes in layer IIi (10) and with horizontally oriented processes in layer IId (5) were seen and were reminiscent of autoradiographic labeling patterns obtained previously following tectal injection of tritiated GABA. In the cerebellum, GABA-like immunoreactivity involved all types of neurons with the exception of granule cells. Purkinje cells showed regionally different intensities of immunostaining. In addition, in folium X no stained basket-like elements were observed. Although there is no evidence as yet about the function of GABA in most of the structures, the present results indicate an important role for this neurotransmitter in the pigeon brain.

Strain differences in catecholamine content of pigeon brains

Concentrations of dopamine, noradrenaline and DOPAC were measured in 6 regions of telencephalon and in the cerebellar cortex of adult white Carneaux, and mixed breed pigeons of either sex. In both groups the regional differences in the amount of dopamine replicated the pattern we published earlier. The absolute values, however, differed in these groups: in all telencephalic regions the amount of dopamine was lower in the Carneaux strain than in the mixed breed specimens; in 4 of these regions the difference was statistically significant. A similar tendency was observed in the noradrenaline concentration but the difference was significant only in two telencephalic samples and the cerebellum. The concentration of DOPAC was significantly smaller in two telencephalic regions of the Carneaux pigeons.

Neurotransmitter receptors in the avian brain. II. Muscarinic cholinergic receptors

The characteristics and distribution of muscarinic cholinergic receptors were examined in the pigeon brain using in vitro receptor autoradiography. The antagonist N-[ 3 H] methylscopolamine was used as ligand and the presence of the putative subtypes M 1 and M 2 of the muscarinic cholinergic receptors examined using carbachol and pirenzepine as displacers. The highest densities of muscarinic cholinergic receptors in the pigeon brain were localized in the paleostriatum augmentatum and the lobus parolfactorius, areas homologous to the mammalian corpus striatum. In contrast, the paleostriatum primitivum, corresponding to the mammalian globus pallidus, was poor in muscarinic cholinergic receptors. The rest of the telencephalon was also rich in muscarinic cholinergic receptors, while thalamic, hypothalamic and brainstem areas, as well as the tectum, presented intermediate densities, similar to the mammalian brain. Exceptions to that were the hippocampus, which was poorly labeled in the bird brain and the cerebellum, which presented intermediate to high densities of muscarinic cholinergic receptors in the bird brain. Preliminary pharmacological studies suggest differences between avian and mammalian receptor subtypes, since carbachol and pirenzepine apparently did not recognize two different receptor subpopulations.

Neurotransmitter receptors in the avian brain. I. Dopamine receptors

The distribution of dopamine D 1 and D 2 receptors was investigated in the pigeon brain using in vitro receptor autoradiography. D 1 receptors were labeled in slide-mounted tissue sections with [ 3H]SCH 23390. The agonist [ 3H]CV 205–502 and the antagonist [ 3H]spiroperidol were used to label D 2 receptors. The highest densities of both D 1 and D 2 receptors were concentrated in the paleostriatum augmentatum, lobus paraolfactorius and olfactory tubercle. D 1 receptors were in addition enriched in the entire pigeon telencephalon with exception of the ectostriatum. In contrast, no significant densities of D 2 receptors were observed in the pallium. D 2 but not D 1 receptors were present in the tectum and nucleus pretectalis. The cerebellum presented D 1 but not D 2 receptors. These results demonstrate that in the avian brain, like in the mammalian one, both dopamine D 1 and D 2 receptors are concentrated in ‘striatal’ areas, while the telencephalon is rich in D 1 but not D 2 receptors.