Inhibition of brain NOS activity impair spatial learning acquisition in fish.

Nitric oxide plays a role in the long term potentiation mechanisms produced in the mammalian hippocampus during spatial learning. A great deal of data has demonstrated that the dorsolateral telencephalon of fish could be homologous to the mammalian hippocampus sharing functional similarities. In the present study, we analyzed the role of nitric oxide in spatial learning in teleost fish. In Experiment 1, we studied the effects of the inhibition of telencephalic nitric oxide in goldfish during the acquisition of a spatial task. The results showed that nitric oxide is involved in the learning of a spatial task. Experiment 2 evaluated the effects of the inhibition of telencephalic nitric oxide in goldfish for the retrieval of a learned spatial response. The results indicated that the retrieval of the information previously stored is not dependent of the nitric oxide. The last experiment analyzed the role of the telencephalic nitric oxide in place and cue learning. Results showed a clear impairment in place but not in cue learning. As a whole, these results indicate that fish and mammals, could have a relational memory system mediated by similar biochemical mechanisms.

Oleoylethanolamide and Palmitoylethanolamide Protect Cultured Cortical Neurons Against Hypoxia.

Introduction: Perinatal hypoxic-ischemic (HI) encephalopathy is defined as a neurological syndrome where the newborn suffers from acute ischemia and hypoxia during the perinatal period. New therapies are needed. The acylethanolamides, oleoylethanolamide (OEA) and palmitoylethanolamide (PEA), possess neuroprotective properties, and they could be effective against perinatal HI. These lipid mediators act through peroxisome proliferator-activated receptors subtype α (PPARα), or transient receptor potential vanilloid (TRPV), such as TRPV subtype 1 and 4. Materials and Methods: The objectives of this study were to discern: (1) the neuroprotective role of OEA and PEA in parietotemporal cortical neurons of newborn rats and mice subjected to hypoxia, and (2) the role of the receptors, PPARα, TRPV1, and TRPV4, in neuroprotective effects. Cell culture of cortical neurons and the lactate dehydrogenase assay was carried out. The role of receptors was discerned by using selective antagonist and agonist ligands, as well as knockout (KO) PPARα mice. Results: The findings indicate that OEA and PEA exert neuroprotective effects on cultured cortical neurons subjected to a hypoxic episode. These protective effects are not mediated by the receptors, PPARα, TRPV1, or TRPV4, because neither PPARα KO mice nor receptor ligands significantly modify OEA and PEA-induced effects. Blocking TRPV4 with RN1734 is neuroprotective per se, and cotreatment with OEA and PEA is able to enhance neuroprotective effects of the acylethanolamides. Since stimulating TRPV4 was devoid of effects on OEA and PEA-induced protective effects, effects of RN1734 cotreatment seem to be a consequence of additive actions. Conclusion: The lipid mediators, OEA and PEA, exert neuroprotective effects on cultured cortical neurons subjected to hypoxia. Coadministration of OEA or PEA, and the TRPV4 antagonist RN1734 is able to enhance neuroprotective effects. These in vitro results could be of utility for developing new therapeutic tools against perinatal HI.

Palmitoylethanolamide prevents neuroinflammation, reduces astrogliosis and preserves recognition and spatial memory following induction of neonatal anoxia-ischemia.

RATIONAL: Neonatal anoxia-ischemia (AI) particularly affects the central nervous system. Despite the many treatments that have been tested, none of them has proven to be completely successful. Palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) are acylethanolamides that do not bind to CB1 or CB2 receptors and thus they do not present cannabinoid activity. These molecules are agonist compounds of peroxisome proliferator-activator receptor alpha (PPARα), which modulates the expression of different genes that are related to glucose and lipid metabolism, inflammation, differentiation and proliferation. OBJECTIVE: In the present study, we analyzed the effects that the administration of PEA or OEA, after a neonatal AI event, has over different areas of the hippocampus. METHODS: To this end, 7-day-old rats were subjected to AI and then treated with vehicle, OEA (2 or 10 mg/kg) or PEA (2 or 10 mg/kg). At 30 days of age, animals were subjected to behavioral tests followed by immunohistochemical studies. RESULTS: Results showed that neonatal AI was associated with decreased locomotion, as well as recognition and spatial memory impairments. Furthermore, these deficits were accompanied with enhanced neuroinflammation and astrogliosis, as well as a decreased PPARα expression. PEA treatment was able to prevent neuroinflammation, reduce astrogliosis and preserve cognitive functions. CONCLUSIONS: These results indicate that the acylethanolamide PEA may play an important role in the mechanisms underlying neonatal AI, and it could be a good candidate for further studies regarding neonatal AI treatments.

Animal Models of Maladaptive Traits: Disorders in Sensorimotor Gating and Attentional Quantifiable Responses as Possible Endophenotypes.

Traditional diagnostic scales are based on a number of symptoms to evaluate and classify mental diseases. In many cases, this process becomes subjective, since the patient must calibrate the magnitude of his/her symptoms and therefore the severity of his/her disorder. A completely different approach is based on the study of the more vulnerable traits of cognitive disorders. In this regard, animal models of mental illness could be a useful tool to characterize indicators of possible cognitive dysfunctions in humans. Specifically, several cognitive disorders such as schizophrenia involve a dysfunction in the mesocorticolimbic dopaminergic system during development. These variations in dopamine levels or dopamine receptor sensibility correlate with many behavioral disturbances. These behaviors may be included in a specific phenotype and may be analyzed under controlled conditions in the laboratory. The present study provides an introductory overview of different quantitative traits that could be used as a possible risk indicator for different mental disorders, helping to define a specific endophenotype. Specifically, we examine different experimental procedures to measure impaired response in attention linked to sensorimotor gating as a possible personality trait involved in maladaptive behaviors.

Neural basis of the spatial navigation based on geometric cues.

Recent studies have found that hippocampus of mammals and birds and the lateral pallium of the fish telencephalon are critical for learning the geometric properties of space. Nevertheless, other studies suggest that navigation based on geometric information is primarily supported by proximal cues near the target location. According to this hypothesis, animals could use a taxon strategy to navigate an environment where only geometric cues are available and the results from lesion studies could be masking other effects related to the use of featural information. In the present study, we examined the effects of lesions to the lateral pallium of goldfish in the encoding of geometric spatial information. Goldfish with telencephalic lesions were trained to search for a goal in a rectangular-shaped arena with either one or two possible goals. Lateral pallium lesions do not interfere with goal location when the geometric information defined the goal unambiguously. Present results suggest that the geometric information is sensitive to be encoded in taxon strategies and therefore it could not depend directly on the correct functioning of the hippocampal system.

What are the functions of fish brain pallium?

There is some experimental evidence that the pallial areas of a fish's brain are involved in distincted learning functions. Recently published data suggest that the medial pallium is essential for avoidance learning and the lateral pallium is crucial for spatial learning and is also involved in temporal aspects of the learning processes. This data joined to the proposal of homologies between medial and lateral fish pallia and pallial amygdala and hippocampus respectively, suggest that the pallial areas could have preserved their functions throughout vertebrates' evolution. However, the functional implication of dorsal pallium that has been proposed as homologous to mammalian isocortex and transition cortex is largely unknown. In this study we analyze the role of dorsal pallium in trace and non-trace avoidance learning. Our results show the implication of this area in trace conditioning, but not in non-trace conditioning. This result allows discussion of homology proposals among lateral, dorsal, and medial pallia and hippocampus, isocortex, and pallial amygdala respectively.

Age-related increase in the immunoproteasome content in rat hippocampus: molecular and functional aspects.

Alterations in the proteasome activity in the CNS have been described during aging. However, a detailed study of all proteasome subunits is actually lacking. We have analyzed, in vivo, the age-related modifications in the molecular composition of hippocampal proteasomes. We found that the immunoproteasome/proteasome ratio was increased in aged hippocampus. The processing of the low-molecular-mass protein (LMP)7/beta(5i) subunit, practically absent in young hippocampus, was increased in aged animals. Among the potential factors underlying these modifications we evaluated the neuroinflammation and the transcription factor Zif268. Lipopolysaccharide (LPS)-induced neuroinflammation in young rats, up-regulated the expression of immunoproteasome subunits and increased the processing of the LMP7/beta(5i) protein. Moreover, the hydrophobicity of cellular peptides, analyzed by liquid chromatography, increased in both, young LPS-injected animals and aged rats, suggesting that immunoproteasomes including the LMP7/beta(5i) subunit could, at least in part, account for this modification. Also, the mRNA expression of the transcription factor Zif268, which down-regulates the immunoproteasome subunit LMP7/beta(5i) by binding to sequences within the promoter regions, was decreased in both, aged hippocampus and young LPS-injected animals. Finally, we found that spatial memory training in young animals, a situation in which the expression of Zif268 is increased, modified the mRNA expression of the constitutive and catalytic subunits in an opposite manner. Based on present data, we propose that the age-related increases in the content of hippocampal immunoproteasome is mostly because of neuroinflammatory processes associated to aging.

Role for D-serine within the ventral tegmental area in the development of cocaine's sensitization.

Repeated exposure to cocaine results in motor sensitization that, in the ventral tegmental area (VTA), is associated to enhanced glutamate release, which in turn leads to enhanced calcium levels in dopaminergic neurons. Calcium influx activates calcium-calmodulin-dependent protein kinases such as CaMKII. D-Serine could participate on these effects, and the objective was to discern the role of VTA D-serine after a sensitizing regimen of cocaine (10 mg/kg daily), and to discern consequent expression changes in CaMKII and its activated form. For this purpose, D-serine, sodium benzoate (inhibitor of D-amino acid oxidase, the degradating enzyme of D-serine), and 7-chlorokynurenate (inhibitor of the glycine site of NMDA receptors) were injected into the VTA (in either the induction or expression phase of sensitization), and activation state of CaMKII was assessed through blotting. The findings indicated that intra-VTA administration of D-serine (5 mM) and sodium benzoate (100 and 200 microg/microl) during the induction phase (not expression) reliably augmented the expression of behavioral sensitization to cocaine, providing evidence that D-serine in the VTA participates in the initiation of motor sensitization to this psychostimulant drug. Intra-VTA infusions of D-serine, sodium benzoate and 7-chlorokynurenate did not elicit a motor effect of their own. Confirming the important role of NMDA receptors and their activation at the glycine site, the employment of 7-chlorokynurenate (2 and 5 microg/microl) led to blocking of the development of sensitization to cocaine. CaMKII within the VTA was found to participate in D-serine's effects because this kinase, that is activated after repeated cocaine, was further activated after co-treatment with D-serine or sodium benzoate. Besides CaMKII activity was otherwise reduced by 7-chlorokynurenate.

Telencephalon and geometric space in goldfish.

Neuroanatomical evidence indicates that the lateral pallium (LP) of ray-finned fishes could be homologous to the hippocampus of mammals and birds. Recent studies have found that hippocampus of mammals and birds is critical for learning geometric properties of space. In this work, we studied the effects of lesions to the lateral pallium of goldfish on the encoding of geometric spatial information. Goldfish with telencephalic lesions were trained to search for a goal in a rectangular-shaped arena containing one different wall that served as the only distinctive environmental feature. Although fish with lateral pallium lesions learned the task even faster than sham and medial pallium (MP)-lesioned animals, subsequent probe trials showed that they were insensitive to geometric information. Sham and medial pallium-lesioned animals could use both geometric and feature information to locate the goal. By contrast, fish with lateral palium lesions relied exclusively on the feature information provided by the wall of a different colour. These results indicate that lesions to the lateral pallium of goldfish, like hippocampal lesions in mammals and birds, selectively impair the encoding of geometric spatial information of environmental space. Thus, the forebrain structures of teleost fish that are neuroanatomically equivalent to the mammalian and avian hippocampus also share a central role in supporting spatial cognition. Present results suggest that the presence of a hippocampal-dependent memory system implicated in the processing of geometric spatial information is an ancient feature of the vertebrate forebrain that has been conserved during the divergent evolution of different vertebrate groups.

Spatial learning and goldfish telencephalon NMDA receptors.

Recent results have demonstrated that the mammalian hippocampus and the dorso-lateral telencephalon of ray-finned fishes share functional similarities in relation to spatial memory systems. In the present study, we investigated whether the physiological mechanisms of this hippocampus-dependent spatial memory system were also similar in mammals and ray-finned fishes, and therefore possibly conserved through evolution in vertebrates. In Experiment 1, we studied the effects of the intracranial administration of the noncompetitive NMDA receptor antagonist MK-801 during the acquisition of a spatial task. The results indicated dose-dependent drug-induced impairment of spatial memory. Experiment 2 evaluated if the MK-801 produced disruption of retrieval of a learned spatial response. Data showed that the administration of MK-801 did not impair the retrieval of the information previously stored. The last experiment analyzed the involvement of the telencephalic NMDA receptors in a spatial and in a cue task. Results showed a clear impairment in spatial learning but not in cue learning when NMDA receptors were blocked. As a whole, these results indicate that physiological mechanisms of this hippocampus-dependent system could be a general feature in vertebrate, and therefore phylogenetically conserved.

Emotional and spatial learning in goldfish is dependent on different telencephalic pallial systems.

In mammals, the amygdala and the hippocampus are involved in different aspects of learning. Whereas the amygdala complex is involved in emotional learning, the hippocampus plays a critical role in spatial and contextual learning. In fish, it has been suggested that the medial and lateral region of the telencephalic pallia might be the homologous neural structure to the mammalian amygdala and hippocampus, respectively. Although there is evidence of the implication of medial and lateral pallium in several learning processes, it remains unclear whether both pallial areas are involved distinctively in different learning processes. To address this issue, we examined the effect of selective ablation of the medial and lateral pallium on both two-way avoidance and reversal spatial learning in goldfish. The results showed that medial pallium lesions selectively impaired the two-way avoidance task. In contrast, lateral pallium ablations impaired the spatial task without affecting the avoidance performance. These results indicate that the medial and lateral pallia in fish are functionally different and necessary for emotional and spatial learning, respectively. Present data could support the hypothesis that a sketch of these regions of the limbic system, and their associated functions, were present in the common ancestor of fish and terrestrial vertebrates 400 million years ago.

Lesions of the medial pallium, but not of the lateral pallium, disrupt spaced-trial avoidance learning in goldfish (Carassius auratus).

The effects of telencephalic lesions of the medial pallium (MP) and lateral pallium (LP) of goldfish on avoidance learning were studied in a two-way, shuttle response, spaced-trial avoidance conditioning situation. Animals received one trial per day, a training regime that permits the assessment of avoidance learning in the absence of stimulus carry-over effects from prior trials. Control and LP-lesioned goldfish exhibited significantly faster avoidance learning than MP-lesioned animals. These results suggest that the MP, but not the LP, is responsible for the widely described deficits in avoidance learning after lesions of the entire telencephalon. The proposal of a functional similarity between the fish MP and the mammalian amygdala, known to be involved in fear conditioning, suggests a conservative phylogenetic role of this area in avoidance learning.

Avoidance response in goldfish: emotional and temporal involvement of medial and lateral telencephalic pallium.

The hippocampus and the amygdala are involved in avoidance learning in mammals. The medial and lateral pallia of actinopterygian fish have been proposed as homologous to the mammalian pallial amygdala and hippocampus, respectively, on the basis of neuroanatomical findings. This work was aimed at studying the effects of ablation of the medial telencephalic pallia (MP) and lateral telencephalic pallia (LP) in goldfish on the retention of a conditioned avoidance response previously acquired in two experimental conditions. In the first experiment, fish were trained in nontrace avoidance conditioning. In the second experiment, fish were trained in trace avoidance conditioning in which temporal cues were crucial for the learning process. An MP lesion affected the retention of the avoidance response in both procedures; in contrast, an LP lesion impaired the retention only in the trace-conditioning procedure. These data support the presence of two different systems of memory in fish, based on discrete telencephalic areas: the MP, involved in an emotional memory system; and the LP, involved in a spatial, relational, or temporal memory system. Moreover, these differential effects were similar to those produced by amygdalar and hippocampal lesions in mammals. We conclude that these specialized systems of memory could have appeared early during phylogenesis and could have been conserved throughout vertebrate evolution.

Involvement of the telencephalon in spaced-trial avoidance learning in the goldfish (Carassius auratus).

Goldfish (Carassius auratus) received escape-avoidance training in a shuttle-response situation at a rate of a single trial per day. Widely spaced training evaluates the ability of a discriminative stimulus to control an avoidance response in the absence of stimulus carry-over effects from prior recent trials. In Experiment 1, master goldfish exhibited significantly faster avoidance learning than yoked controls. The results suggest that the shuttle response was instrumentally acquired. Experiment 2 demonstrated a significant deficit in the acquisition of avoidance behavior following ablation of the telencephalon. The implications of spaced-trial, telencephalon-dependent avoidance learning, as demonstrated in these experiments for the first time, are discussed in the context of comparative research on instrumental learning in goldfish. These results provide further support for the hypothesis that the fish telencephalon contains an emotional system that is critical for fear conditioning.