Trnka Vaclav - Central European Anatomist and Medical Polymath of the Eighteenth Century.

Vaclav Trnka from Krovice (1739-1791, in Latin: Wenzel Trnka Krzowitz) was a remarkable physician whose life serves as an example in the history of medicine by connecting major capital cities of Central Europe. In view of current geographical layout, he was born and brought up in the Czech Republic, graduated from University of Vienna in Austria, and was appointed Professor of the Anatomy at the newly established Faculty of Medicine of University of Nagyszombat, presently Trnava in Slovak Republic. When the University moved to Buda and later to Pest (today Budapest, Hungary), he was the first educator to introduce anatomy as a medical subject to be taught in a Hungarian medical school. He also was elected the Dean of Faculty of Medicine three times and in 1786-1787 he acted as Rector of then the Royal University of Pest. During his life, he published twenty-seven monographs dealing with different areas of clinical medicine, such as malaria (intermittent fever), diabetes, and rickets. Based on these monographs we can proclaim that Václav Trnka was a co-founder of modern infectology, diabetology and ophthalmology in Central Europe. Nowadays, artificial intelligence and bioinformatics are inseparable parts of modern health care system which help the transformation of big data into valuable knowledge. In the 18th century, Professor Trnka owned more than 3,000 scientific books and had natural, innate intelligence and wisdom which made him a real "medical polymath". As a musician, Trnka also composed sixty-one canons, two of them long wrongly considered as Mozart's work. Despite the fact that Trnka is considered to be the founder of Hungarian anatomy education and a major medical figure of the eighteenth century Central Europe, no internationally acclaimed biographical record of his life or work has so far been published in English. Therefore, we would like to reintroduce Václav Trnka both as an anatomist and medical polymath, and to give an overview of the early days of anatomy teaching in present-day Slovakia and Hungary (Fig. 1, Ref. 27). Keywords: Trnka from Krovice, anatomist, medical polymath, history of medicine.

Receptors of peptides as therapeutic targets in epilepsy research.

Neuropeptides are signaling molecules participating in the modulation of synaptic transmission. Neuropeptides are stored in dense core synaptic vesicles, the release of which requires profound excitation. Only in the extracellular space, neuropeptides act on G-protein coupled receptors to exert a relatively slow action both pre- and postsynaptically. Consequently, neuropeptide modulators are ideal candidates to influence epileptic tissue overexcited during seizures. Indeed, a number of neuropeptides have receptors implicated in epilepsy and many of them are considered to participate in endogenous neuroprotective actions. Neuropeptide receptors, present in the hippocampus, the most frequent focus of seizures in temporal lobe epilepsy, received the largest attention as potential anti-epileptic targets. Receptors of hippocampal neuropeptides, somatostatin, neuropeptide Y, galanin, dynorphin, enkephalin, substance P, cholecystokinin, vasoactive intestinal polypeptide, and receptors of some neuropeptides, which are also hormones such as ghrelin, angiotensins, corticotropin- releasing hormone, adrenocorticotropin, thyrotropin-releasing hormone, oxytocin and vasopressin involved in epilepsy are discussed in the review article. Activation and inhibition of receptors by oral application of peptides as drugs is typically not efficient because of low bioavailability: rapid degradation and insufficient penetration of peptides through the blood-brain barrier. Recent progress in the development of non-peptide agonists and antagonists of neuropeptide receptors as well as gene therapeutic approaches leading to the local production of agonists and antagonists within the central nervous system will also be discussed.

Microvascular regulatory role and increased expression of vascular endothelial growth factor receptor type 2 in experimental gingivitis.

OBJECTIVE: The aim of the present study was to investigate the possible microvascular regulatory role of vascular endothelial growth factor receptor type 2 (VEGFR2) in experimental gingivitis in rats. BACKGROUND: Our previous results demonstrated that functionally active VEGFR2s are located in the venules of rat gingiva. While there is no remarkable endogenous gingival VEGF production under normal circumstances, exogenous VEGF, via VEGFR2, shows venodilatory effects. We assumed that VEGF plays an important role in vasoregulatory processes (vasodilation, increased permeability, angiogenesis) of gingival inflammation. METHODS: Gingivitis was induced by placing ligatures and composite material around and between the lower incisors of anesthetized Wistar rats next to the gingival margin. Seven days later, VEGFR2 antagonist (ZM323881), was dripped upon the labial gingiva next to the lower incisors. Diameter changes of the selected gingival venules were measured by vital microscopy. Animals with healthy gingiva served as controls. Venule diameter changes were compared to the baseline and to control groups (no ligature). Immunohistochemical and Western blot analysis for VEGFR2 were utilized. RESULTS: After 15, 30 and 60 min of local application of ZM323881, there was a significant venoconstriction in the inflamed gingiva compared to the baseline, while no change was recorded in controls. Endothelium, smooth muscle cells and pericytes of the gingivitis group showed increased VEGFR2 expression. CONCLUSION: Our findings suggest that there is an increased VEGF production in gingivitis, which may play an important role in vasodilation of rat gingival venules.

Antiinflammatory effect of BPC 157 on experimental periodontitis in rats.

The pentadecapeptide BPC 157 has been shown to have anti-inflammatory and wound healing effects on multiple target tissues and organs. The purpose of the present study was to investigate the effect of BPC 157 on inflammation and bone resorption in experimental periodontitis in rats. First the acute effect of BPC was tested on gingival blood flow by laser doppler flowmetry. Then periodontitis was produced by a silk ligature placed around the lower left first molar. Rats were treated with BPC 157 (once daily for 12 days) or vehicle. At day 13, the gingivomucosal tissues encircling the molars were removed on both sides. Inflammation was assessed by Evans blue plasma extravasation technique and by histology. Alveolar bone loss was analyzed by microCT. BPC 157 had no effect on gingivomucosal blood flow. Twelve day ligature caused a significantly increased Evans blue extravasation in the gingivomucosal tissue, histological signs of inflammation, and alveolar bone destruction. BPC 157 treatment significantly reduced both plasma extravasation, histological alterations and alveolar bone resorption. In conclusion, systemic application of BPC 157 does not alter blood circulation in healthy gingiva. Chronic application of the peptide has potent antiinflammatory effects on periodontal tissues in ligature induced periodontitis in rats. Taken together, this proof of concept study suggests that BPC 157 may represent a new peptide candidate in the treatment of periodontal disease.

Evidence for a role of GABA interneurones in the cortical modulation of midbrain 5-hydroxytryptamine neurones.

Recent electrophysiological studies demonstrate that the ventral medial prefrontal cortex has a powerful inhibitory influence on 5-hydroxytryptamine (5-HT) neurones in the dorsal raphe nucleus. Here we utilised a combination of anatomical and electrophysiological methods to characterise the cellular substrate underlying this effect.Anterograde tracing (Phaseolus vulgaris leucoagglutinin) using electron microscopy demonstrated a pathway from the ventral medial prefrontal cortex that makes neuronal contacts throughout the dorsal raphe nucleus. These contacts were predominantly asymmetrical synapses adjoining GABA immunoreactive dendrites and spines. In vivo extracellular recordings were made in the dorsal raphe nucleus of the anaesthetised rat from a subpopulation of non-5-HT neurones. These neurones were fast-firing, irregular and with short spike width, properties strongly reminiscent of immunochemically identified GABA interneurones in other brain regions. Recordings of classical 5-HT neurones were also included. Electrical stimulation of the ventral medial prefrontal cortex elicited a rapid onset (16 ms latency), orthodromic excitation of the non-5-HT neurones (13/25 neurones). This stimulation also caused a pronounced inhibition of most 5-HT neurones tested, with a longer latency (30 ms), and this was partially blocked by locally applied bicuculline. These data provide the first evidence that the ventral medial prefrontal cortex influences the activity of large numbers of raphe 5-HT neurones by targeting a local network of GABA neurones. This circuitry predicts that physiological and pathological changes in the ventral medial prefrontal cortex will impact on significant parts of the forebrain 5-HT system.

Evidence for reactive nitrogen species formation in the gingivomucosal tissue.

An increase in nitric oxide production has been demonstrated in periodontitis. Here we investigated the potential role of nitric-oxide-derived nitrating species (such as peroxynitrite) in a rat model of ligature-induced periodontitis. Formation of 3-nitrotyrosine, the stable product formed from tyrosine reacting with nitric-oxide-derived nitrating species, was detected in the gingivomucosal tissue. 3-Nitrotyrosine immunohistochemical analysis revealed a significant elevation in the number of immunopositive leukocytes, and higher immunoreactivity of the gingival ligaments and epithelium in the ligated than in the contralateral (control) side. On both sides, several 3-nitrotyrosine-positive bands and, on the ligated side, a unique 52-kDa 3-nitrotyrosine-positive band were detected by Western blot. However, in the sterile gingivomucosal tissue of rat pups, no 3-nitrotyrosine or inducible nitric oxide synthase immunoreactivity was found. Analysis of these data suggests that resident bacteria of the gingivomucosal tissue induce an increase in reactive nitrogen species, which is greatly enhanced by plaque formation in periodontitis.

Evidence for the expression of cyclooxygenase-2 enzyme in periodontitis.

We investigated the role of the inducible isoform of cyclooxygenase (COX-2) in a rat model of periodontitis using a selective COX-2 inhibitor NS-398. Periodontitis was produced by a silk ligature placed around the lower left 1st molar. Animals were treated with NS-398 (3 mg kg(-1) i.p., 2 times per day for 7 days) or vehicle. At Day 8, the gingivomucosal tissues encircling the mandibular 1st molars were removed on both sides for COX-2 immunohistochemistry, measurement of plasma extravasation by the Evans blue technique, and alveolar bone loss by videomicroscopy. Immunohistochemical analysis revealed numerous strongly COX-2-positive cells in the subepithelial tissues in the ligated side and only a few COX-2-reactive cells in the contralateral (control) side. Ligation significantly increased Evans blue extravasation in the gingivomucosal tissue and alveolar bone destruction compared to the control side. NS-398 treatment significantly reduced the plasma extravasation and alveolar bone resorption of the ligated side compared to vehicle administration. The present results suggest that COX-2 is induced by periodontitis, and plays an important role in gingival inflammation and alveolar bone destruction. In a previous study (Br J Pharmacol 1998;123:353-60) we found the expression of the inducible isoform of nitric oxide synthase in this model. Therefore, based on our own data and the literature, we propose that selective inhibition of these inducible enzymes might be a basis for adjunctive therapy, or new therapeutic approaches in periodontitis.

Changes in binding to muscarinic and nicotinic cholinergic receptors in the chick telencephalon, following passive avoidance learning.

Changes in nicotinic and muscarinic cholinergic receptors 30 min after one-trial passive avoidance training were studied in day-old chicks (Gallus domesticus), by quantitative receptor autoradiography. [3H]-alpha-bungarotoxin (alpha-BgT) and [3H]-quinuclidinyl benzilate (QNB) were used to monitor changes in 15 forebrain regions for nicotinic and muscarinic receptors, respectively. A significant increase occurred bilaterally in the quantity of bound alpha-BgT in the lobus parolfactorius, while the amount of bound QNB decreased significantly, and bilaterally, in the hippocampus, hyperstriatum ventrale, lobus parolfactorius and posterolateral telencephalon, pars dorsalis. The data support an involvement of cholinergic receptor types in the neural mechanisms underlying passive avoidance learning.

The avian hippocampal formation: subdivisions and connectivity.

The avian hippocampal formation (HP) is considered to be homologous to the mammalian hippocampus on the basis of topography, developmental origin and its role in processing spatial memory. However, the morphological organization of the avian HP is very different from that of mammals and components similar to the subdivisions of the mammalian structure are not readily recognizable. In passerine birds, three spatially and morphologically distinct populations of Calbindin immunoreactive neurones are found in the dorsolateral (DL), dorsomedial (DM) and ventral (V) aspects of HP. Iontophoresis of Phaseolus vulgaris leucoagglutinin revealed three consistently different projection patterns arising from the different subregions. Generally, there is a medial-to-lateral topographical organization of efferents in relation to the septal complex. The DL region could be paralleled to the subiculum of mammals with its main projections to the basal ganglia, the limbic archistriatum, the lateral septum and the paraxial meso-diencephalic centres. The 'V' subdivision is likely to be homologous to the Ammon's horn of mammals with its commissural projections to the contralateral HP. Based on its purely intrinsic connectivity, the DM region could be a good candidate for an equivalent of the dentate gyrus. Nitric oxide synthase (NOS) containing neural structures display a specific distribution within the hippocampal subregions which is uniform in all passerine species studied. However, there is a marked difference in the level of diffuse neuropil reactivity between food-storers versus non-storers. Unlike the mammalian homologue, avian hippocampal NOS positive neurones do not show a near complete co-localization with the inhibitory transmitter GABA.

Distribution of glial fibrillary acidic protein and vimentin-immunopositive elements in the developing chicken brain from hatch to adulthood.

The present study describes the distribution of glial fibrillary acidic protein (GFAP) and vimentin-immunopositive structures in the brain of the domestic chicken (Gallus domesticus) from hatching to maturity. The telencephalon is penetrated by a vimentin-immunopositive radial fibre system, representing a modified form of radial glia, in day-old chicks. Numerous fibres of this system persist until adulthood, mainly in the lobus parolfactorius, lamina medullaris dorsalis and lamina frontalis superior. GFAP immunoreactivity also appears in the course of development in these fibres. The distribution of GFAP-immunopositive astrocytes in the post-hatch telencephalon is like that found in adult chicken, except for the ectostriatum, in which an adult-like GFAP-immunostaining only develops during week three. This delay may be associated with a relatively slow maturation of this visual centre. In the diencephalon and in the mesencephalic tegmentum of day-old chicks GFAP-immunopositive astrocytes are confined to the border zone of several nuclei. In these areas as well as in the pons most GFAP positive astrocytes only appear gradually during the first two post-hatch weeks, although radial fibres occur only sparsely at hatch. Summarizing these results, a gradual replacement of radial fibres by astrocytes, typical of mammals, cannot be found in chicken. In the nucleus laminaris we observed a characteristic palisade of non-ependymal glia, reactive to GFAP but not to vimentin, which almost completely disappears by adulthood. We suggest that this glial system is instrumental in the development of the dendritic organisation of this nucleus. The optic tectum displays a dense array of GFAP-immunopositive radial glia at hatching, similar in this to the situation found in reptiles. However, in the tectum of reptiles this radial glia persists for the lifetime, whereas in the chick it disappears from the superficial tectal layers. This phenomenon may reflect the fact that there is no replacement of tectal cells or regeneration of retinotectal pathways in the chicken. In the early stage, the large cerebral tracts were found to contain dense accumulations of GFAP-positive cells, with peculiarly long outgrowths accompanying nerve fibres. No vimentin-immunopositivity was found in these glial elements; however vimentin was present in the glia situated at the optic chiasm, the anterior commissure and at other decussations. These structures, as well as the raphe, displayed the most intense vimentin-immunopositivity in the post-hatch chicken. This characteristic glial population may represent glial elements that have been reported to regulate fibre-crossing at the midline.

An electrophysiological and neuroanatomical study of the medial prefrontal cortical projection to the midbrain raphe nuclei in the rat.

In this study we utilized electrophysiological and pathway tracing methods to investigate the projections from the medial prefrontal cortex to the midbrain raphe nuclei of the rat. Initial pathway tracing experiments using retrograde (horseradish peroxidase conjugates with wheatgerm agglutinin or choleratoxin B subunit) and anterograde (Phaseolus vulgaris-leucoagglutinin) markers demonstrated a direct, bilateral projection to the dorsal raphe nucleus and median raphe nucleus from the medial prefrontal cortex, and the origin of this projection was localized predominantly in the ventral medial prefrontal cortex (infralimbic/dorsal penduncular cortices). Using chloral hydrate-anaesthetized rats, extracellular recordings were made mostly from 5-hydroxytryptamine neurons in the dorsal raphe nucleus, but non-5-hydroxytryptamine dorsal raphe neurons were also studied, as was a small number of 5-hydroxytryptamine neurons in the median raphe nucleus. In an initial study, electrical stimulation of the ventral medial prefrontal cortex caused a post-stimulus inhibition in the majority (49/56) of dorsal raphe 5-hydroxytryptamine neurons tested (mean duration of inhibition, 200+/-17 ms); in some cases (8/56) the inhibition was preceded by short-latency (26 +/-3 ms) orthodromic activation, and a small number of cells was antidromically activated (6/56). Both single spiking and burst-firing 5-hydroxytryptamine neurons in the dorsal raphe nucleus responded in the same way, and median raphe 5-hydroxytryptamine neurons were also inhibited (5/5). In contrast, few (2/12) of the non-5-hydroxytryptamine dorsal raphe neurons tested were inhibited by ventral medial prefrontal cortex stimulation. The effects of stimulation of the dorsal and ventral medial prefrontal cortex were compared on the same raphe 5-hydroxytryptamine neurons (n=17): ventral medial prefrontal cortex stimulation inhibited 16/17 of these neurons while only 8/17 were inhibited by dorsal medial prefrontal cortex stimulation. Finally, the inhibitory effect of ventral medial prefrontal cortex stimulation on 5-hydroxytryptamine cell-firing was not altered by 5-hydroxytryptamine depletion with p-chlorophenylalanine or by systemic administration of the selective 5-hydroxytryptamine1A receptor antagonist WAY 100635. The latter findings indicate that the inhibition is not due to release of raphe 5-hydroxytryptamine which could theoretically arise from anti- or orthodromically activated 5-hydroxytryptamine neurons. Our results show that stimulation of the ventral medial prefrontal cortex causes a marked post-stimulus inhibition in the vast majority of midbrain raphe 5-hydroxytryptamine neurons tested. It seems likely that the projection from ventral medial prefrontal cortex to the midbrain raphe nuclei mediates the responses of 5-hydroxytryptamine neurons to cortical stimulation. These data are relevant to recent discoveries of functional and structural abnormalities in the medial prefrontal cortex of patients with major depressive illness.

Nitric oxide synthase containing nerves in the cat and dog dental pulp and gingiva.

In a previous study we found that nitric oxide (NO) plays an essential role in the hemodynamic regulation of the feline dental pulp. However, no evidence for the presence of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) containing nerve fibers was found in the rat and cat dental pulps. In the present study, we are first to report the presence of a small number of NADPH-d positive and/or NO synthase immunoreactive perivascular and solitary varicose axons in the dental pulp and abundant number of similar axons in the gingiva of cats and dogs. These fibres may travel within the inferior alveolar nerve and might participate in sensory (i.e. pain) as well as in autonomic (i.e. regulation of blood flow) innervation of the dental pulp and gingiva.

Synaptic terminals immunolabelled against glutamate in the lobus parolfactorius of domestic chicks (Gallus domesticus) in relation to afferents from the archistriatum.

The lobus parolfactorius (LPO) has been implicated in memory formation associated with passive avoidance training of young posthatch domestic chicks. The anatomical circuitry underlying memory formation in the chick is likely to involve the intermediate medial hyperstriatum ventrale-archistriatum-LPO arc. In the present work, we attempted to combine an ultrastructural characterisation of archistriatal afferent terminals in LPO with a description of the synaptic structure of LPO, in particular those elements that are immunoreactive to glutamate and GABA. Ventral archistriatal regions of 7-day-old domestic chicks were iontophoretically injected with Phaseolus vulgaris leucoagglutinin and the anterograde transport of the tracer was detected in the LPO. Selected samples from these birds, and also from other day-old chicks, were resin-embedded and reacted for L-glutamate or GABA, using the postembedding immunocytochemical method. Glutamate was abundant in the neuropil of LPO and typically seen in axodendritic or axospinous terminals with asymmetrical junctions, often multiple or perforated postsynaptic appositions. Conversely, GABA was often present in aspinous dendrites, probably representing GABAergic local circuit neurons or (putative striatonigral) projection neurons. Archistriatal efferents terminating in LPO formed small en passant or terminal varicosities, with infrequent asymmetrical axospinous synapses. Glutamate was not detected in these boutons. The findings imply that the functional state of LPO, based on powerful glutamatergic excitation, may be modified by a non-glutamatergic archistriatal input.

Efferent connections of the domestic chick archistriatum: a phaseolus lectin anterograde tracing study.

The archistriatum of the domestic chick has been implicated in both fear behaviour and learning. However, relatively little is known about its organisation. The efferent connections of discrete anatomical regions of the chick archistriatum were therefore investigated by iontophoresis of the anterograde tracer Phaseolus vulgaris leucoagglutinin into its anterior, dorsal intermediate, ventral intermediate, medial, and posterior parts. The results of this study suggest that the chick archistriatum can be divided into two basic divisions according to whether they project to the following limbic structures: the hippocampal formation, septal areas, lobus parolfactorius, nucleus accumbens, ventral paleostriatum, and dorsomedial thalamus. The limbic archistriatum includes the posterior archistriatum and extends rostrally through the ventral intermediate archistriatum into the anterior archistriatum. The non-limbic archistriatum comprises the dorsal intermediate and medial archistriatum and largely gives rise to specific sensory, somatosensory, and motor telencephalofugal efferents. There may not be distinct borders between these two divisions of the chick archistriatum.

Efferent connectivity of the hippocampal formation of the zebra finch (Taenopygia guttata): an anterograde pathway tracing study using Phaseolus vulgaris leucoagglutinin.

The avian hippocampal formation (HP) is considered to be homologous to the mammalian hippocampus, being involved in memory formation and spatial memory in particular. The subdivisions and boundaries of the pigeon hippocampus have been defined previously by various morphological methods to detect further similarities with the mammalian homologue. We studied the efferent projections of the zebra finch hippocampus by applying Phaseolus vulgaris leucoagglutinin, and three main subdivisions were distinguished on the basis of the connectivity patterns. Dorsolateral injections gave rise to projections innervating the rostralmost extension of the HP, a laminar complex including the dorsal and ventral hyperstriata and the lamina frontalis superior, the rostral lobus parolfactorius, the medial and ventral paleostriatal regions, the lateral septal nucleus, the nucleus of the diagonal band, the dorsolateral corticoid area, the archistriatum posterius, and the nucleus taeniae in the telencephalon. In the diencephalon, labelled axons were seen in the periventricular and lateral hypothalamus, including the lateral mammillary nuclei, and in the dorsolateral and the dorsomedial posterior thalamic nuclei, whereas, in the midbrain, only the area ventralis of Tsai contained hippocampal fibres. With the exception of the bilateral archistriatal efferents, all projections were ipsilateral. Dorsomedial injections gave rise to a local fibre system that was almost completely restricted to the ipsilateral hippocampal formation. In addition, lectin-containing fibres continued in the dorsal septal region and a thin band in the hyperstriatum accessorium, adjacent to the lateral ventricle. Ventral injections gave rise to axons innervating ipsilaterally the dorsolateral subdivision, and bilaterally the medial septal nuclei and the contralateral ventral hippocampus.

Distribution of nitric oxide synthase containing elements in the feline submandibular gland.

Combined nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunocytochemistry were used to study the distribution of nitric oxide synthesizing elements in the cat submandibular gland. A large number of thin varicose fibres, with intense staining for both markers, were seen around or in close contact with the acini. Some of the stained nerve fibres were associated with intra- and interlobular salivary ducts and blood vessels. All neurones in the submandibular ganglia showed intense staining for both NADPH-d and NOS. The epithelial layer of the salivary ductal branches and the endothelial lining of blood vessels were NOS immunonegative but NADPH-d positive. Our results suggest that NO might act as a neurotransmitter in the regulation of blood flow and secretion in the submandibular salivary gland.

Connectivity of the lobus parolfactorius of the domestic chicken (Gallus domesticus): an anterograde and retrograde pathway tracing study.

In 1-week-old domestic chicks, the connectivity of the lobus parolfactorius (LPO), part of the avian basal ganglia, was investigated using Phaseolus vulgaris leucoagglutinin and horseradish peroxidase for anterograde and retrograde pathway tracing, respectively. Tyrosine hydroxylase immunocytochemistry was applied in combination with Phaseolus lectin to assess the overlap between LPO efferents and diencephalic and mesencephalic catecholamine centres. Anterograde projections from LPO were detected in the hyperstriatum, neostriatum, and paleostriatum. Intranuclear connections were also apparent within the LPO. Descending LPO efferents innervated the lateral mammillary and intramedial nuclei and the dorsomedial thalamic complex. Fibres from LPO were observed in the tectal gray, substantia nigra, area ventralis tegmentalis of Tsai, and the adjacent nucleus mesencephalicus profundus. Further caudally, projections from LPO reached the nucleus papillioformis, locus coeruleus, and subcoeruleus ventralis. LPO efferents were coextensive with tyrosine hydroxylase-positive cells in the nuclei mamillaris lateralis and intramedialis of the hypothalamus, area ventralis tegmentalis, substantia nigra, locus coeruleus, and subcoeruleus ventralis of mesencephalic and pontine tegmentum. Close contacts between LPO fibres and catecholamine cells were visible in the nigra and the area ventralis tegmentalis. Retrograde labelling from LPO was found in the archistriatum, dorsomedial thalamic complex, nuclei lateralis anterior and superficialis parvicellularis thalami, substantia nigra, central gray, area ventralis tegmentalis of Tsai, and locus coeruleus and in cells dorsal to the decussation of brachium conjunctivum. Reciprocal connections were verified between the LPO and the following areas: dorsomedial thalamic complex, central gray, substantia nigra, area ventralis of Tsai, and locus coeruleus.

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 of 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.

Transient cerebral ischemia disrupts performance on a one-trial passive avoidance task in the domestic chick and is associated with neuronal degeneration in the central nervous system.

We have examined the effects of transient cerebral ischemia on performance of a one-trial passive avoidance task by chicks. Transient forebrain ischemia was induced by bilateral carotid artery occlusion for a period of 10 min. In one experimental group, ischemia was produced prior to training on the avoidance task whereas in the other group ischemic intervention was not made until 3 h after initial training. Sham-operated groups were matched to each of the experimental groups. All four groups were tested for retention of the avoidance response 24 h post-surgery. The sham-operated birds and those receiving post-training ischemia showed good retention of the avoidance response, whereas in birds which received ischemia prior to training there was significant amnesia. Neuronal damage, determined qualitatively using a silver impregnation method, was observed in several forebrain regions including the hippocampus, hyperstriatal regions, paleostriatum primitivum, ventral archistriatum, and lateral corticoid area. Damage was also observed in the Purkinje cells of the cerebellum. The behavioural and anatomical effects of transient forebrain ischemia have not been previously investigated in an avian species and the finding of significant amnesia for a learning task following ischemia is in good agreement with several behavioural studies in mammals.

Quantitative autoradiographic demonstration of changes in binding to delta opioid, but not mu or kappa receptors, in chick forebrain 30 minutes after passive avoidance training.

Day-old domestic chicks (Gallus domesticus) were trained on a one-trial passive avoidance task in which the aversive stimulus was a bitter tasting substance, methylanthranilate. Thirty minutes later, localization of binding of highly specific ligands (([D-Ala2, Gly-ol]-enkephalin ([3H]DAGO) for mu (mu) receptor sites, [D-Pen2,D-Pen5]-enkephalin ([3H]-DPDPE) for delta (delta) sites, and [3H]-U- 69593 for kappa (kappa 1) sites) to opioid receptors in various regions of the forebrain of methyl-anthranilate trained (M-) and control (water trained (W-)) chicks was determined using quantitative receptor autoradiography. Significant differences in binding to delta ([3H]-DPDPE), but not mu or kappa receptors, were found in several regions of the forebrain, of trained compared to control chicks. There were decreases in binding in the hyperstriatum dorsale of the left hemisphere (14%) and a decrease in binding in the lateral hyperstriatum ventrale of the right hemisphere (14%). However, significant increases were observed in delta binding in the paleostriatum augmentatum of the right hemisphere (16%) and the lobus parolfactorius of both hemispheres (left, 20%; right, 21%). In a control experiment designed to determine whether the taste of methylanthranilate contributed to the increase in 3H-DPDPE binding, there was no significant difference in the level of binding between blindfolded birds in which methylanthranilate was placed in the beak, and blindfolded birds in which water was placed on the bead and inserted into the beak. These findings demonstrate that changes occur in an opioid receptor sub-type in specific regions of forebrain of the chick following passive avoidance training which may be related to events concerned with the process of memory formation.