Ágota Ádám

Efferent Connections of Septal Nuclei of the Domestic Chick (Gallus domesticus): An Anterograde Pathway Tracing Study with a Bearing on Functional Circuits

Small iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin were placed in different subregions of the septum of domestic chicks. The main targets of septal projections comprised the ipsi‐ and contralateral septal nuclei, including the nucleus of the diagonal band, basal ganglia, including the ventral paleostriatum, lobus parolfactorius, nucleus accumbens, and olfactory tubercle, archistriatum, piriform cortex, and anterior neostriatum. Further diencephalic and mesencephalic septal projections were observed in the ipsilateral preoptic region, hypothalamus (the main regions of afferentation comprising the lateral hypothalamic nuclei, ventromedial, paraventricular and periventricular nuclei, and the mammillary region), dorsal thalamus, medial habenular and subhabenular nuclei, midbrain central gray, and ventral tegmental area. Contralateral projections were also encountered in the septal nuclei, ventral paleostriatum, periventricular and anteromedial hypothalamic nuclei, suprachiasmatic nucleus, and the lateral hypothalamic area. Avian septal efferents are largely similar to those of mammals, the main differences being a relatively modest hippocampal projection arising mainly from the nucleus of the diagonal band (as confirmed by a specific experiment with the retrograde pathway tracer True blue), the lack of interpeduncular projection, and a greater contingent of amygdalar efferents arising from the lateral septum rather than the nucleus of the diagonal band. This pattern of connectivity is likely to reflect an important role of the avian septal nuclei in the coordination of limbic circuits and the integration of a wide variety of information sources modulating the appropriate behavioral responses: attention and arousal level, memory formation, hormonally mediated behaviors, and their affective components (such as ingestive, reproductive, and parental behaviors), social interaction, locomotor modulation, and circadian rhythm. J. Comp. Neurol. 469:437–456, 2004.

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.

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 ∼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. J. Comp. Neurol. 498:266–276, 2006.

Abundance and location of DARPP-32 in striato-tegmental circuits of domestic chicks.

The striatum is reciprocally connected to the brainstem dopaminergic nuclei and receives a strong dopaminergic input. In the present study the spatial relation between the dopaminergic and dopaminoceptive structures of the avian medial striatum (formerly: lobus parolfactorius) was observed by confocal laser scanning microscope in the domestic chick (Gallus domesticus). We also analysed the connections in the area ventralis tegmentalis and the substantia nigra. To label the dopaminergic structures, anti-tyrosine hydroxylase was used and DARPP-32 (dopamine and cAMP regulated phosphoprotein) was a marker of dopaminoceptive elements. The tyrosine hydroxylase positive fibres formed baskets of juxtapositions around the DARPP-32 containing cells of the medial striatum. However, such baskets were also observed to juxtapose DARPP-32 immunonegative cells. In the tegmentum, DARPP-32 was observed in axons descending from the telencephalon via the ansa lenticularis. These varicose fibers innervated the ventral tegmental area and substantia nigra and were often juxtaposed to dopaminergic neurons and dendrites. Approximately 40% of the striatal projection neurons targeting the ventral tegmentum, and 60% of striatal projection neurons targeting the nigra were immunoreactive to DARPP-32, as revealed by retrograde pathway tracing with Fast Blue. Endogenous dopamine may exert a retrograde synaptic effect on the afferent striato-tegmental fibers, apart from the reported extrasynaptic action. The abundance of juxtapositions observed in the avian brainstem and medial striatum corroborates the possibility of reciprocal striato-tegmental circuits, relevant to the reinforcement of behaviour.

Apoptotic effects of the 'designer drug' methylenedioxypyrovalerone (MDPV) on the neonatal mouse brain

The designer drug of cathinone family, methylenedioxypyrovalerone (MDPV), is a cheap and frequently used psychoactive drug of abuse. However, its mechanism of action, particularly its potential detrimental effect on the developing brain, is largely unknown, despite the fact that pregnant females may occur among the users. The objective of our study was to identify the brain areas sensitive for a possible apoptotic effect of the widely abused MDPV on the developing brain. To this end, we used a mouse model which can be compared with the human fetus of third trimester, considering the developmental stage of the brain. Litters of 7-day-old C57BL/6J mice were treated either with i.p. injection of 10mg/kg b.wt.of MDPV or vehicle (saline), and sacrificed after 24h. Similar dose of MDPV enhanced locomotor activity of pups. The brains were processed for anti-caspase 3 (Casp3) immunohistochemistry and the apoptotic cells were identified and counted. We found prominent increase in the number of apoptotic cells in the piriform cortex, retrosplenial area, hippocampus CA1 and nucleus accumbens, whereas the overall density of cells did not change significantly in these regions. The neurons of the nucleus accumbens appeared to be especially sensitive to MDPV: Casp3-immunoreactive cells marked out the core and shell regions of the accumbens. Highest percentage of apoptotic cells as compared to total cell density was also found in the nucleus accumbens. However, we did not observe the same effect on the brain of adult mice. Thus, MDPV did not seem to increase apoptosis in the mature nervous system. The results are in agreement with the assumption that cathinones (in particular MDPV) may adversely affect neural integrity in the developing CNS.

The cannabinoid CB1 receptor antagonist rimonabant dose-dependently inhibits memory recall in the passive avoidance task in domestic chicks (Gallus domesticus).

The effect of endocannabinoids on synaptic plasticity has been demonstrated in a variety of species and brain regions. Relatively little is known about the localization and significance of cannabinoid (CB) receptors in the avian brain. The objective of the present study was to investigate the effect of a specific CB(1) receptor antagonist upon the acquisition and consolidation of memory in young domestic chicks. One-day-old domestic chicks (Gallus domesticus) were trained and tested by the passive avoidance paradigm. Systemic (i.p.) administration of the CB(1) receptor antagonist rimonabant in a dose of 1mg/kg 30 min before the training failed to affect learning, but a similar treatment 30 min before the recall (5.5h after training) attenuated the retention in 60% of animals. In another set of animals, a dose of 0.01 mg/kg produced no significant impairment, whereas doses 0.1mg/kg and 1.0mg/kg resulted in significant attenuation in passive avoidance performance when tested 30 min prior to recall. The results are discussed in terms of a putative mediating role of CB receptors in the consolidation of memory.

Amygdalofugal axon terminals immunoreactive for L-aspartate or L-glutamate in the nucleus accumbens of rats and domestic chickens: A comparative electron microscopic immunocytochemical study combined with anterograde pathway tracing

Several studies have shown that L-aspartate (Asp) is present in synaptic vesicles and released exocytotically from presynaptic terminals, possibly by Ca2+-dependent corelease of Asp and L-glutamate (Glu). It has been demonstrated that both excitatory amino acids (EAAs) are released from the rat striatum as part of corticostriatal neurotransmission. The single or colocalized occurrence of Asp and Glu in specific synaptic boutons of the chicken medial striatum/nucl. accumbens has been demonstrated by our group using ultrastructural immunocytochemistry. However, evidence for the presence of EAAs in any specific striatal pathway was only circumstantial. Here, we report on the distribution of Asp and Glu in specific synaptic terminals of the amygdalostriatal pathway, both in rat and chicken brains, combining anterograde tracing with postembedding immunogold labeling of Asp or Glu. Immunoreactivity for Asp and Glu was observed in amygdalofugal terminals with asymmetrical synaptic junctions (morphologically representing excitatory synapses) in both species. The postsynaptic targets were either dendritic spines or small dendrites, whereas axosomatic or axo-axonic connections were not observed. Ultrastructurally, the synaptic terminals immunoreactive for Asp were indistinguishable from those immunoreactive for Glu. The findigs are consistent with an Asp–Glu corelease mechanism, with a distinct synaptic contingent, evolutionarily conserved in the amygdalostriatal pathway.

Effect of synthetic cathinones: mephedrone, butylone and 3,4 methylene-dioxypyrovalerone (MDPV) on social separation induced distress vocalization, vigilance and postural control of young domestic chicks.

Designer drugs have become a distinct social problem and health hazard in Europe and US, and their abuse has increased dramatically in the last decade. Selective effects of these agents on animal behavioral parameters may help in better understanding of the potential risks of human drug abuse. In the present study, the effects of three different abusive agents of the cathinone family, mephedrone, butylone and 3,4 methylene-dioxypyrovalerone (MDPV) were tested in young domestic chicks, following administration of single intraperitoneal injections (10mg/bwt). Early maturing (precocial) birds are particularly suited for investigation of isolation stress-related behavioral response and stereotypic or targeted pecking. Both mephedrone and MDPV increased the frequency of distress calls of socially isolated birds as measured over a period of 10min. While this effect of mephedrone was only evident in the first half of observation period, an increase with MDPV was more lasting. Though increased non-distress vocalization, butylone failed to enhance distress calls probably due to a general adverse effect on muscle tone. Apart from its effect on distress vocalization, mephedrone did not alter the behavior of chicks. However, both butylone and MDPV showed prominent behavioral changes, which were examined in another set of long term experiments, over a period of 120min. Butylone caused hyperventilation and a robust impairment of postural control, whereas neither the wakeful activity level, nor the pecking frequency was significantly affected. Conversely, no hyperventilation or postural disorder was observed with MDPV, however, both waking state and pecking were significantly enhanced. The results may be relevant to potentially different and specific effects of cathinone drugs under stress-related conditions, as well as on other physiological and behavioral parameters, even in case of closely related compounds.

Gestational exposure to the synthetic cathinone methylenedioxypyrovalerone results in reduced maternal care and behavioral alterations in mouse pups

The member of synthetic cathinone family, methylenedioxypyrovalerone (MDPV), is a frequently used psychoactive drug of abuse. The objective of our study was to determine the effect of MDPV (administered from the 8th to the 14th day of gestation) on the behavior of neonatal and adolescent mice, as well as its effect on maternal care. We measured maternal care (pup retrieval test, nest building), locomotor activity (open field test), and motor coordination (grip strength test) of dams, whereas on pups we examined locomotor activity at postnatal day 7 and day 21 (open field test) and motor coordination on day 21 (grip strength test). On fresh-frozen brain samples of the dams we examined the expression of two important peptides implicated in the regulation of maternal behavior and lactation: tuberoinfundibular peptide 39 (TIP39) mRNA in the thalamic posterior intralaminar complex, and amylin mRNA in the medial preoptic nucleus. We detected decreased birth rate and survival of offspring, and reduced maternal care in the drug-treated animals, whereas there was no difference between the motility of treated and control mothers. Locomotor activity of the pups was increased in the MDPV treated group both at 7 and 21 days of age, while motor coordination was unaffected by MDPV treatment. TIP39 and amylin were detected in their typical location but failed to show a significant difference of expression between the drug-treated and control groups. The results suggest that chronic systemic administration of the cathinone agent MDPV to pregnant mice can reduce birth rate and maternal care, and it also enhances motility (without impairment of motor coordination) of the offspring.

A dizájner drog metiléndioxi-pirovaleron hatása a fejlődő idegrendszerre kísérletes állatmodellben

Absztrakt Bevezetes: A szintetikus kationok csaladjaba tartozo „dizajner drog”, a metilendioxi-pirovaleron gyakran hasznalt pszichoaktiv szer. Celkitűzes: A szerzők arra a kerdesre kerestek valaszt, hogyan hat a metilendioxi-pirovaleron a vemhesseg 8. es 14. napja kozott adva az utodegerek kozponti idegrendszerenek fejlődesere es a viselkedesukre. Modszer: Nősteny egereket a vemhesseg ezen időszakaban 1×10 mg/ttkg metilendioxi-pirovaleron-oldattal kezeltek subcutan, a kontrollcsoport fiziologias sooldatot kapott. Mertek az anyaallatok utodgondozasi hajlandosagat, lokomotoros aktivitasat es motoros koordinaciojat. Az utodokon a postnatalis 7. es 21. napon ugyanezeket a teszteket vegeztek el. Eredmenyek: Az anyaallatok utodgondozasi hajlandosaga csokkent. A lokomotoros aktivitasi tesztben nem volt kulonbseg a kronikusan kezelt allatok es a kontrollcsoport kozott. A motoros koordinacios teszt eredmenyei alapjan a kronikusan kezelt allatok motoros koordinacioja rosszabb volt. A metilendioxi-pirovaleronnal kro...