Ghrelin treatment leads to dendritic spine remodeling in hippocampal neurons and increases the expression of specific BDNF-mRNA species.

Ghrelin (Gr) is an orexigenic peptide that acts via its specific receptor, GHSR-1a distributed throughout the brain, being mainly enriched in pituitary, cortex and hippocampus (Hp) modulating a variety of brain functions. Behavioral, electrophysiological and biochemical evidence indicated that Gr modulates the excitability and the synaptic plasticity in Hp. The present experiments were designed in order to extend the knowledge about the Gr effect upon structural synaptic plasticity since morphological and quantitative changes in spine density after Gr administration were analyzed "in vitro" and "in vivo". The results show that Gr administered to hippocampal cultures or stereotactically injected in vivo to Thy-1 mice increases the density of dendritic spines (DS) being the mushroom type highly increased in secondary and tertiary extensions. Spines classified as thin type were increased particularly in primary extensions. Furthermore, we show that Gr enhances selectively the expression of BDNF-mRNA species.

Contextual learning and context effects during infancy: 30 years of controversial research revisited.

Over the last 30years a considerable number of reports have explored learning about context during infancy in both humans and rats. This research was stimulated by two different theoretical frameworks. The first, known as the neuromaturational model, postulates that learning and behavior are context-independent during early ontogeny, a hypothesis based on the idea that contextual learning is dependent on the hippocampal function, and that this brain structure does not reach full maturity until late in infancy. The second theoretical framework views infants not as immature organisms, but rather as perfectly matured ones, given that their behavioral and cognitive capacities allow them to adapt appropriately to the demands of their specific environment in accordance with their maturational level. This model predicts significant ontogenetic variations in learning and memory due to developmental differences in what is perceived and attended to during learning episodes, which can result in ontogenetic differences in contextual learning depending on the specific demands of the task. The present manuscript reviews those studies that have examined potential developmental differences in contextual learning and context effects in rats. The reviewed results show that, during infancy, context can exert a similar influence over learning and memory as that described for the adult rat. Moreover, in some cases, contextual learning and context effects were greater in infants than in adults. In contrast, under other experimental conditions, no evidence of contextual learning or context effects was observed. We analyzed the procedural factors of these studies with the aim of detecting those that favor or impede contextual learning during infancy, and we discussed whether existing empirical evidence supports the claim that the functionality of the hippocampus is a limiting factor for this type of learning during infancy. Finally, conclusions from human research into contextual learning capacities during infancy were also examined. In view of the wealth of evidence showing contextual learning and context effects during infancy, we suggest that future research aimed at exploring the involvement of the hippocampus in this type of learning should be conducted using parameters which allow the expression of contextual learning during each ontogenetic period.

Context-dependent latent inhibition in preweanling rats.

Preexposure to a conditioned stimulus (CS) usually weakens conditioning, an effect known as latent inhibition. Similar to other learning interference effects, latent inhibition has been characterized as context-dependent, which means that the magnitude of this effect can be attenuated by changing the context between the different phases of the procedure (e.g., preexposure and conditioning). Latent inhibition has been found with a variety of procedures in infant rats, but the few studies that examined the context-dependency of this phenomenon during this ontogenetic period found no context-change effect. The present study explored the context-dependency of latent inhibition during infancy using a conditioned taste aversion preparation and employing contexts enriched with distinctive odors to increase the possible efficacy of the context manipulation. Experiment 1 showed that three preexposures to the CS (saccharin) were sufficient to retard conditioning to the same CS, although this effect was also observed in a control group preexposed to an alternative taste stimulus (saline), in comparison with a non-preexposed control group. In Experiment 2a, the CS-preexposure effect was found to be specific to the preexposed CS when the number of preexposures was increased. This effect was revealed as context-dependent in Experiment 2b, since it was attenuated by changing the context between preexposure and conditioning. The present result is consistent with recent studies showing the context-dependency of extinction in preweanling rats, thus demonstrating these animals' capacity to learn about context early on in their development.

Spontaneous recovery from extinction in the infant rat.

Within the Pavlovian conditioning framework, extinction is a procedure in which, after conditioning, the conditioned stimulus (CS) is repeatedly presented without the unconditioned stimulus (US). During this procedure the conditioned response (CR) is gradually attenuated. It has been suggested that extinction during the early stages of ontogeny is a qualitatively different process from extinction in adulthood: during infancy, extinction may result in erasure of the memory, while during adulthood extinction involves new learning. This conclusion was supported by studies showing that renewal, reinstatement or spontaneous recovery procedures were not effective during infancy for recovering the CR once it had been extinguished. These studies used the freezing response as the only behavioral index, although some recent evidence indicates that the absence of freezing after conditioning or after extinction does not necessarily imply a deficit in memory, and that other behavioral indexes may be more sensitive to detecting conditioning effects. The goal of the present study was to analyze extinction in preweanling rats by examining the possibility of the spontaneous recovery of a conditioned fear response, measured through a different set of mutually-exclusive behaviors that constitute an exhaustive ethogram, and including control groups (Experiment 1: US-Only and CS-Only; Experiment 2: US-Only, CS-Only and Unpaired) in order to examine whether non-associative learning may explain quantitative or qualitative changes in the frequency of specific responses during extinction or recovery. Extinction produced changes in the expression of freezing, grooming and exploration, and the clearest evidence of spontaneous recovery came from the analysis of freezing behavior. The pattern of behavior observed during extinction is compatible with theoretical approaches which consider different dynamic behavioral systems, and it also fit in well with a molar approach to the analysis of behavior, which considers that extinction involves a transition from one allocation of time among behaviors to another allocation, rather than a loss of strength in any particular discrete response. These results have implications for the study of extinction during infancy, since they are compatible with the hypothesis that the original memory survives extinction, and highlight the importance of control conditions for detecting this effect during this ontogenetic period.

A sensory-enhanced context allows renewal of an extinguished fear response in the infant rat.

Studies of extinction in preweanling rats have failed to find ABA-renewal in a fear conditioning paradigm. This result supports the hypothesis postulating ontogenetic qualitative differences in experimental extinction. A similar result in adult rats led to the conclusion that ABA-renewal requires contexts A and B to differ in several types of features, including odor cues. Recently we reported experimental evidence of the renewal of an extinguished taste aversion response in infant rats employing contexts which differ in their odor content. The present study examines the possibility of renewing an extinguished fear response in infant rats when contexts A and B do not include (Experiment 1) or include (Experiment 2) an explicit odor. Results showed absence of renewal when using standard contexts (without explicit odors, Experiment 1). However, when contexts A and B varied also in their odor content, the ABA-renewal procedure was effective in reinstating the extinguished CR (Experiment 2). Thus, it can be concluded that the sensory content of the context determines the observation of renewal in the infant rat, a result that is coherent with previous observations in the adult rat. As a whole, these results challenge our understanding of extinction as a learning process that is qualitatively different in preweanling rats than in later stages of ontogeny.

Fibronectin substrate induces shortening of in vitro neural crest cell mitotic cycle

Neural crest cells (NCC) segregate from neural folds, swim through extracellular matrix, and differentiate in several cell types. During in vivo and in vitro dispersion, NCC display a substantial proliferative rate. Here, we have evaluated this biological behavior under the influence of different substrates. NCC exhibit the highest proliferative activity on fibronectin substrate, with shortening of the G1 period. This fact is reverted by specific antibody pre-treatment of the substrate. Taking into account that NCC migrate along narrow and restricted pathways rich in essential fibronectin, with high proliferative rates, these results indicate that the fibronectin pathway contributes to increase the cell number, leading to a high population pressure that could participate in the early dispersion of NCC

Fibronectin substrate induces shortening of in vitro neural crest cell mitotic cycle

Neural crest cells (NCC) segregate from neural folds, swim through extracellular matrix, and differentiate in several cell types. During in vivo and in vitro dispersion, NCC display a substantial proliferative rate. Here, we have evaluated this biological behavior under the influence of different substrates. NCC exhibit the highest proliferative activity on fibronectin substrate, with shortening of the G1 period. This fact is reverted by specific antibody pre-treatment of the substrate. Taking into account that NCC migrate along narrow and restricted pathways rich in essential fibronectin, with high proliferative rates, these results indicate that the fibronectin pathway contributes to increase the cell number, leading to a high population pressure that could participate in the early dispersion of NCC

Fibronectin substrate induces shortening of in vitro neural crest cell mitotic cycle.

Neural crest cells (NCC) segregate from neural folds, swim through extracellular matrix, and differentiate in several cell types. During in vivo and in vitro dispersion, NCC display a substantial proliferative rate. Here, we have evaluated this biological behavior under the influence of different substrates. NCC exhibit the highest proliferative activity on fibronectin substrate, with shortening of the G1 period. This fact is reverted by specific antibody pre-treatment of the substrate. Taking into account that NCC migrate along narrow and restricted pathways rich in essential fibronectin, with high proliferative rates, these results indicate that the fibronectin pathway contributes to increase the cell number, leading to a high population pressure that could participate in the early dispersion of NCC.

Fibronectin substrate induces shortening of in vitro neural crest cell mitotic cycle.

Neural crest cells (NCC) segregate from neural folds, swim through extracellular matrix, and differentiate in several cell types. During in vivo and in vitro dispersion, NCC display a substantial proliferative rate. Here, we have evaluated this biological behavior under the influence of different substrates. NCC exhibit the highest proliferative activity on fibronectin substrate, with shortening of the G1 period. This fact is reverted by specific antibody pre-treatment of the substrate. Taking into account that NCC migrate along narrow and restricted pathways rich in essential fibronectin, with high proliferative rates, these results indicate that the fibronectin pathway contributes to increase the cell number, leading to a high population pressure that could participate in the early dispersion of NCC.

Method for treating and processing whole chick embryos for autoradiography, immunocytochemistry and other techniques.

A method is presented for in situ treatment of whole chick embryos with drugs and immunocytochemical and fixative reagents that resembles conditions "in ovo." The chick embryo is placed in a "shell-less" culture system where it is contained by an agar ring allowing for treatment in vivo. The conceptus (embryo+membranes) is then mounted on a microporous membrane and inserted into a filter device connected to a three-way stopcock that permits fluids to be changed using syringes. The embryo is then processed in toto or after embedding and sectioning for light or electron microscopy. The proposed handling system decreases technical artifacts and changes in the topographic microanatomy produced by conventional manipulation of chick embryos. This method is useful also for directly observing and recording changes in the embryo during drug treatments and allows processing with dangerous reagents without their direct contact with the operator. It is simple, inexpensive and requires only minimal technical training.