Facial nerve axotomy induces morphological changes in hippocampal pyramidal neurons.

Facial nerve injury in rats have been widely used to study functional and structural changes that occur in the injured motoneurons and other central nervous system structures related with sensorimotor processing. A decrease in long-term potentiation of hippocampal CA3-to-CA1 commissural synapse has recently been reported related to this peripheral injury. Additionally, it has been found increased corticosterone plasmatic levels, impairment in spatial memory consolidation, and hippocampal microglial activation in animals with facial nerve axotomy. In this work, we analyzed the neuronal morphology of hippocampal CA1 and CA3 pyramidal neurons in animals with either reversible or irreversible facial nerve injury. For this purpose, brain tissues of injured animals sacrificed at different postlesion times, were stained with the Golgi-Cox method and compared with control brains. It was found that both reversible and irreversible facial nerve injury-induced significant decreases in dendritic tree complexity, dendritic length, branch points, and spine density of hippocampal neurons. However, such changes' timing varied according to hippocampal area (CA1 vs. CA3), dendritic area (apical vs. basal), and lesion type (reversible vs. irreversible). In general, the observed changes were transient when animals had the possibility of motor recovery (reversible injury), but perdurable if the recovery from the lesion was impeded (irreversible injury). CA1 apical and CA3 basal dendritic tree morphology were more sensible to irreversible injury. It is concluded that facial nerve injury induced significant changes in hippocampal CA1 and CA3 pyramidal neurons morphology, which could be related to LTP impairments and microglial activation in the hippocampal formation, previously described.

Facial nerve injury-associated hippocampal microglial activation / Activación de la microglía en el hipocampo asociada con lesión del nervio facial

Introduction: Facial nerve injury induces changes in hippocampal long-term synaptic plasticity and affects both object recognition memory and spatial memory consolidation (i.e., hippocampus-dependent tasks). Although facial nerve injury-associated microglial activation has been described regarding the primary motor cortex, it has not been ascertained whether something similar occurs in the hippocampus. Peripheral nerve injuryassociated microglial changes in hippocampal tissue could explain neuronal changes in the contralateral hippocampus. Objective: To characterize the effect of unilateral facial nerve injury on microglial proliferation and activation in the contralateral hippocampus. Materials and methods. Immunohistochemical experiments detected microglial cells in the hippocampal tissue of rats that had undergone facial nerve injury. The animals were sacrificed at specific times after injury to evaluate hippocampal microglial cell proliferation (cell density) and activation (cell area); sham-operated animals were compared to lesioned animals sacrificed 1, 3, 7, 21, or 35 days after injury. Results: Facial nerve-injured rats' hippocampal microglial cells proliferated and adopted an activated phenotype 3- to 21-days post-lesion. Such modifications were transient since the microglial cells returned to their resting state five weeks after injury, despite the injury's irreversible nature. Conclusions: Facial nerve injury causes the transient proliferation and activation of microglial cells in the hippocampus. This finding might partly explain the morphological and electrophysiological changes described for CA1 pyramidal neurons and the impairment of spatial memory consolidation which has previously been observed in facial nerve-injured rats.

Introducción. Las lesiones del nervio facial afectan la plasticidad a largo plazo en el hipocampo, así como la memoria de reconocimiento de objetos y la memoria espacial, dos procesos dependientes de esta estructura.  Objetivo. Caracterizar en ratas el efecto de la lesión unilateral del nervio facial sobre la activación de células de la microglía en el hipocampo contralateral. Materiales y métodos. Se hicieron experimentos de inmunohistoquímica para detectar células de la microglía en el hipocampo de ratas sometidas a lesión irreversible del nervio facial. Los animales se sacrificaron en distintos momentos después de la lesión, para evaluar la evolución de la proliferación (densidad de células) y la activación (área celular) de la microglía en el tejido del hipocampo. Los tejidos cerebrales de los animales de control se compararon con los de animales lesionados sacrificados en los días 1, 3, 7, 21 y 35 después de la lesión. Resultados. Las células de la microglía en el hipocampo de animales con lesión del nervio facial mostraron signos de proliferación y activación a los 3, 7 y 21 días después de la lesión. Sin embargo, al cabo de cinco semanas, estas modificaciones se revirtieron, a pesar de que no hubo recuperación funcional de la parálisis facial. Conclusiones. La lesión irreversible del nervio facial produce proliferación y activación temprana y transitoria de las células de la microglía en el hipocampo. Estos cambios podrían estar asociados con las modificaciones electrofisiológicas y las alteraciones comportamentales dependientes del hipocampo descritas recientemente.

Activación de la microglía en el hipocampo asociada con lesión del nervio facial / Facial nerve injury-associated hippocampal microglíal activation

Introducción. Las lesiones del nervio facial afectan la plasticidad a largo plazo en el hipocampo, así como la memoria de reconocimiento de objetos y la memoria espacial, dos procesos dependientes de esta estructura. Si bien se ha descrito una activación de la microglía en la corteza motora primaria asociada con esta lesión, no se conoce si ocurre algo similar en el hipocampo. Objetivo. Caracterizar en ratas el efecto de la lesión unilateral del nervio facial sobre la activación de células de la microglía en el hipocampo contralateral. Materiales y métodos. Se hicieron experimentos de inmunohistoquímica para detectar células de la microglía en el hipocampo de ratas sometidas a lesión irreversible del nervio facial. Los animales se sacrificaron en distintos momentos después de la lesión, para evaluar la evolución de la proliferación (densidad de células) y la activación (área celular) de la microglía en el tejido del hipocampo. Los tejidos cerebrales de los animales de control se compararon con los de animales lesionados sacrificados en los días 1,3, 7, 21 y 35 después de la lesión. Resultados. Las células de la microglía en el hipocampo de animales con lesión del nervio facial mostraron signos de proliferación y activación a los 3, 7 y 21 días después de la lesión. Sin embargo, al cabo de cinco semanas, estas modificaciones se revirtieron, a pesar de que no hubo recuperación funcional de la parálisis facial. Conclusiones. La lesión irreversible del nervio facial produce proliferación y activación temprana y transitoria de las células de la microglía en el hipocampo. Estos cambios podrían estar asociados con las modificaciones electrofisiológicas y las alteraciones comportamentales dependientes del hipocampo descritas recientemente.

Introduction: Facial nerve injury induces changes in hippocampal long-term synaptic plasticity and affects both object recognition memory and spatial memory consolidation (i.e., hippocampus-dependent tasks). Although facial nerve injury-associated microglíal activation has been described regarding the primary motor cortex, it has not been ascertained whether something similar occurs in the hippocampus. Peripheral nerve injury- associated microglíal changes in hippocampal tissue could explain neuronal changes in the contralateral hippocampus. Objective: To characterize the effect of unilateral facial nerve injury on microglíal proliferation and activation in the contralateral hippocampus. Materials and methods: Immunohistochemical experiments detected microglíal cells in the hippocampal tissue of rats that had undergone facial nerve injury. The animals were sacrificed at specific times after injury to evaluate hippocampal microglíal cell proliferation (cell density) and activation (cell area); sham-operated animals were compared to lesioned animals sacrificed 1,3, 7, 21, or 35 days after injury. Results: Facial nerve-injured rats' hippocampal microglíal cells proliferated and adopted an activated phenotype 3- to 21-days post-lesion. Such modifications were transient since the microglíal cells returned to their resting state five weeks after injury, despite the injury's irreversible nature. Conclusions: Facial nerve injury causes the transient proliferation and activation of microglíal cells in the hippocampus. This finding might partly explain the morphological and electrophysiological changes described for CA1 pyramidal neurons and the impairment of spatial memory consolidation which has previously been observed in facial nerve-injured rats.

Facial Nerve Axotomy Induces Changes on Hippocampal CA3-to-CA1 Long-term Synaptic Plasticity.

Peripheral facial axotomy induces functional and structural central nervous system changes beyond facial motoneurons, causing, among others, changes in sensorimotor cortex and impairment in hippocampal-dependent memory tasks. Here, we explored facial nerve axotomy effects on basal transmission and long-term plasticity of commissural CA3-to-CA1 synapses. Adult, male rats were submitted to unilateral axotomy of the buccal and mandibular branches of facial nerve and allowed 1, 3, 7, or 21 days of recovery before performing electrophysiological recordings of contralateral CA3 (cCA3) stimulation-evoked CA1 field postsynaptic potential in basal conditions and after high frequency stimulation (HFS) (six, one-second length, 100 Hz stimuli trains). Facial nerve axotomy induced transient release probability enhancement during the first week after surgery, without significant changes in basal synaptic strength. In addition, peripheral axotomy caused persistent long-term potentiation (LTP) induction impairment, affecting mainly its presynaptic component. Such synaptic changes may underlie previously reported impairments in hippocampal-dependent memory tasks and suggest a direct hippocampal implication in sensorimotor integration in whisking behavior.

Vibrissal paralysis produces increased corticosterone levels and impairment of spatial memory retrieval.

This research was aimed at establishing how the absence of active whisking in rats affects acquisition and recovery of spatial memory. The mystacial vibrissae were irreversibly paralyzed by cutting the facial nerve's mandibular and buccal branches bilaterally in the facial nerve lesion group (N=14); control animals were submitted to sham-surgery (N=15). Sham-operated (N=11) and facial nerve-lesioned (N=10) animals were trained (one session, eight acquisition trials) and tested 24h later in a circular Barnes maze. It was found that facial nerve lesioned-animals adequately acquired the spatial task, but had impaired recovery of it when tested 24h after training as compared to control ones. Plasma corticosterone levels were measured after memory testing in four randomly chosen animals of each trained group and after a single training trial in the maze in additional facial nerve-lesioned (N=4) and sham-operated animals (N=4). Significant differences respecting the elevation of corticosterone concentration after either a single training trial or memory testing indicated that stress response was enhanced in facial nerve-lesioned animals as compared to control ones. Increased corticosterone levels during training and testing might have elicited the observed whisker paralysis-induced spatial memory retrieval impairment.

[Facial nerve injuries cause changes in central nervous system microglial cells].

INTRODUCTION: Our research group has described both morphological and electrophysiological changes in motor cortex pyramidal neurons associated with contralateral facial nerve injury in rats. However, little is known about those neural changes, which occur together with changes in surrounding glial cells. OBJECTIVE: To characterize the effect of the unilateral facial nerve injury on microglial proliferation and activation in the primary motor cortex. MATERIALS AND METHODS: We performed immunohistochemical experiments in order to detect microglial cells in brain tissue of rats with unilateral facial nerve lesion sacrificed at different times after the injury. We caused two types of lesions: reversible (by crushing, which allows functional recovery), and irreversible (by section, which produces permanent paralysis). We compared the brain tissues of control animals (without surgical intervention) and sham-operated animals with animals with lesions sacrificed at 1, 3, 7, 21 or 35 days after the injury. RESULTS: In primary motor cortex, the microglial cells of irreversibly injured animals showed proliferation and activation between three and seven days post-lesion. The proliferation of microglial cells in reversibly injured animals was significant only three days after the lesion. CONCLUSIONS: Facial nerve injury causes changes in microglial cells in the primary motor cortex. These modifications could be involved in the generation of morphological and electrophysiological changes previously described in the pyramidal neurons of primary motor cortex that command facial movements.

Layer 5 Pyramidal Neurons' Dendritic Remodeling and Increased Microglial Density in Primary Motor Cortex in a Murine Model of Facial Paralysis.

This work was aimed at characterizing structural changes in primary motor cortex layer 5 pyramidal neurons and their relationship with microglial density induced by facial nerve lesion using a murine facial paralysis model. Adult transgenic mice, expressing green fluorescent protein in microglia and yellow fluorescent protein in projecting neurons, were submitted to either unilateral section of the facial nerve or sham surgery. Injured animals were sacrificed either 1 or 3 weeks after surgery. Two-photon excitation microscopy was then used for evaluating both layer 5 pyramidal neurons and microglia in vibrissal primary motor cortex (vM1). It was found that facial nerve lesion induced long-lasting changes in the dendritic morphology of vM1 layer 5 pyramidal neurons and in their surrounding microglia. Dendritic arborization of the pyramidal cells underwent overall shrinkage. Apical dendrites suffered transient shortening while basal dendrites displayed sustained shortening. Moreover, dendrites suffered transient spine pruning. Significantly higher microglial cell density was found surrounding vM1 layer 5 pyramidal neurons after facial nerve lesion with morphological bias towards the activated phenotype. These results suggest that facial nerve lesions elicit active dendrite remodeling due to pyramidal neuron and microglia interaction, which could be the pathophysiological underpinning of some neuropathic motor sequelae in humans.

Retracción a largo plazo del árbol dendrítico de neuronas piramidales córtico-faciales por lesiones periféricas del nervio facial / Peripheral facial nerve lesion induced long-term dendritic retraction in pyramidal cortico-facial neurons

Introducción. Poco se sabe sobre las modificaciones morfológicas de las neuronas de la corteza motora tras lesiones en nervios periféricos, y de la implicancia de dichos cambios en la recuperación funcional tras la lesión. Objetivo. Caracterizar en ratas el efecto de la lesión del nervio facial sobre la morfología de las neuronas piramidales de la capa V de la corteza motora primaria contralateral. Materiales y métodos. Se reconstruyeron neuronas piramidales teñidas con la técnica de Golgi-Cox, de animales control (sin lesión) y animales con lesiones y sacrificados a distintos tiempos luego de la lesión. Se utilizaron cuatro grupos: sham (control), lesión 1S, lesión 3S y lesión 5S (animales con lesiones y evaluados 1, 3 y 5 semanas después de la lesión irreversible del nervio facial, respectivamente). Se evaluaron mediante el análisis de Sholl, las ramificaciones dendríticas de las células piramidales de la corteza motora contralateral a la lesión. Resultados. Los animales con lesiones presentaron parálisis completa de las vibrisas mayores durante las cinco semanas de observación. Comparadas con neuronas de animales sin lesiones, las células piramidales córtico-faciales de los lesionados mostraron una disminución significativa de sus ramificaciones dendríticas. Esta disminución se mantuvo hasta cinco semanas después de la lesión. Conclusiones. Las lesiones irreversibles de los axones de las motoneuronas del núcleo facial, provocan una retracción sostenida del árbol dendrítico en las neuronas piramidales córtico-faciales. Esta reorganización morfológica cortical persistente podría ser el sustrato fisiopatológico de algunas de las secuelas funcionales que se observan en los pacientes con parálisis facial periférica.

Introduction. Little evidence is available concerning the morphological modifications of motor cortex neurons associated with peripheral nerve injuries, and the consequences of those injuries on post lesion functional recovery. Objective. Dendritic branching of cortico-facial neurons was characterized with respect to the effects of irreversible facial nerve injury. Materials and methods. Twenty-four adult male rats were distributed into four groups: sham (no lesion surgery), and dendritic assessment at 1, 3 and 5 weeks post surgery. Eighteen lesion animals underwent surgical transection of the mandibular and buccal branches of the facial nerve. Dendritic branching was examined by contralateral primary motor cortex slices stained with the Golgi-Cox technique. Layer V pyramidal (cortico-facial) neurons from sham and injured animals were reconstructed and their dendritic branching was compared using Sholl analysis. Results. Animals with facial nerve lesions displayed persistent vibrissal paralysis throughout the fiveweek observation period. Compared with control animal neurons, cortico-facial pyramidal neurons of surgically injured animals displayed shrinkage of their dendritic branches at statistically significant levels. This shrinkage persisted for at least five weeks after facial nerve injury. Discussion. Irreversible facial motoneuron axonal damage induced persistent dendritic arborization shrinkage in contralateral cortico-facial neurons. This morphological reorganization may be the physiological basis of functional sequelae observed in peripheral facial palsy patients.

[Peripheral facial nerve lesion induced long-term dendritic retraction in pyramidal cortico-facial neurons]. / Retracción a largo plazo del árbol dendrítico de neuronas piramidales córtico-faciales por lesiones periféricas del nervio facial.

INTRODUCTION: Little evidence is available concerning the morphological modifications of motor cortex neurons associated with peripheral nerve injuries, and the consequences of those injuries on post lesion functional recovery. OBJECTIVE: Dendritic branching of cortico-facial neurons was characterized with respect to the effects of irreversible facial nerve injury. MATERIALS AND METHODS: Twenty-four adult male rats were distributed into four groups: sham (no lesion surgery), and dendritic assessment at 1, 3 and 5 weeks post surgery. Eighteen lesion animals underwent surgical transection of the mandibular and buccal branches of the facial nerve. Dendritic branching was examined by contralateral primary motor cortex slices stained with the Golgi-Cox technique. Layer V pyramidal (cortico-facial) neurons from sham and injured animals were reconstructed and their dendritic branching was compared using Sholl analysis. RESULTS: Animals with facial nerve lesions displayed persistent vibrissal paralysis throughout the five week observation period. Compared with control animal neurons, cortico-facial pyramidal neurons of surgically injured animals displayed shrinkage of their dendritic branches at statistically significant levels. This shrinkage persisted for at least five weeks after facial nerve injury. DISCUSSION: Irreversible facial motoneuron axonal damage induced persistent dendritic arborization shrinkage in contralateral cortico-facial neurons. This morphological reorganization may be the physiological basis of functional sequelae observed in peripheral facial palsy patients.

El estrés agudoiinterfiere con la evocación y pomueve la extinción de la memorria espacial en el laberinto de barnes / High stress interfers with evocation and promotes extintion of spatial memory in the Barnes maze

Para evaluar los efectos del estrés agudo sobre la recuperación y la extinción de la memoria espacial, se utilizaron ratas entrenadas en el laberinto circular de Barnes. El entrenamiento consistió de 8 ensayos de adquisición (intervalo entre ensayos, IEE, de 5 min) en donde los animales debían aprender a encontrar una caja meta ubicada en uno de los 18 agujeros del laberinto. Todos los animales adquirieron el aprendizaje espacial, ya que invirtieron menos tiempo en encontrar la caja meta y cometieron menos errores a medida que se sucedían los ensayos de entrenamiento. Veinticuatro horas después del entrenamiento se evaluó la retención y extinción del aprendizaje espacial mediante una prueba con caja meta (PCC) seguida de siete pruebas sin caja (PSC), con un IEE de 5 min. Una hora y media antes de la sesión de evaluación de la memoria un grupo de animales fue sometido a estrés por restricción de movimientos durante una hora, permitiéndoles un período de recuperación de 30 min y otro grupo permaneció en su caja hogar sin manipulación (control). Los resultados indican que el estrés deteriora el proceso de evocación de la memoria espacial, ya que los animales estresados cometieron un mayor número de errores y demoraron más tiempo en encontrar la caja meta durante la PCC , respecto de los controles. Además, el estrés facilita el proceso de extinción, ya que, durante las PSCs los animales estresados no mostraron una persistencia en la exploración del agujero que, en el entrenamiento, conducía a la caja meta.

To evaluate the effects of acute stress on evocation and extinction of a spatial memory task, we used rats trained in the Barnes circular maze. The training protocol consisted of eight acquisition trials (intertrial interval, ITI; 5 min) where animals must learn to find an escape box placed under one of these eighteen holes of the maze. All animals learned the spatial memory task as indicated by diminished escape latency and weighted errors along the eight acquisition trials. Twenty four hours after training spatial memory evocation and extinction were tested (one trial with escape box, and seven consecutive trials without escape box, ITI: 5 min). One hour and a half before memory evaluation session half of the animals underwent movement restriction during one hour (one hour stress, 1H) and were allowed 30 min to recover, while the other half stayed in their home cage without manipulation (control, C). Stressed animals displayed a significant increase both in escape latency and in weighted errors during the trial with scape box. These results indicate that movement restriction-induced stress deteriorates the spatial memory evocation. Moreover, movement restriction-induced stress during one hour facilitates extinction, showed by the non-persistence in the exploration of the escape hole during the trials without escape box.

Vibrissal paralysis unveils a preference for textural rather than positional novelty in the one-trial object recognition task in rats.

In order to explore the role of active whisking in object novelty detection, the performance of rats having bilateral vibrissal paralysis was compared to that of non-lesioned animals in three modified versions of the one-trial object recognition task performed in the dark. Vibrissal paralysis was induced by crushing the buccal and mandibular branches of the facial nerve. Lesioned animals were not different from non-lesioned ones in terms of weight-gain, locomotive activity, motivation to explore, and ability to become habituated to a given environment. Only lesioned animals were unable to discriminate a change in object texture as novelty cue in the first task, designed to test textural novelty detection. In the second task, designed to test positional novelty detection, both lesioned and non-lesioned subjects were able to discriminate a change in object position as novelty cue. In the third task, designed to force the subjects to choose between two conflicting novelty cues (texture and position), non-lesioned subjects displayed a clear-cut preference for textural novelty while subjects having bilateral vibrissal paralysis preferred positional novelty. According to these results, active whisking is necessary for textural, but not for positional novelty detection. Moreover, these results indicate that textural novelty in non-lesioned animals seems to overcome positional novelty if these are in competition in an object recognition memory task.

Histone deacetylase inhibitors improve learning consolidation in young and in KA-induced-neurodegeneration and SAMP-8-mutant mice.

Histone deacetylases (HDAC) are enzymes that maintain chromatin in a condensate state, related with absence of transcription. We have studied the role of HDAC on learning and memory processes. Both eyeblink classical conditioning (EBCC) and object recognition memory (ORM) induced an increase in histone H3 acetylation (Ac-H3). Systemic treatment with HDAC inhibitors improved cognitive processes in EBCC and in ORM tests. Immunohistochemistry and gene expression analyses indicated that administration of HDAC inhibitors decreased the stimulation threshold for Ac-H3, and gene expression to reach the levels required for learning and memory. Finally, we evaluated the effect of systemic administration of HDAC inhibitors to mice models of neurodegeneration and aging. HDAC inhibitors reversed learning and consolidation deficits in ORM in these models. These results point out HDAC inhibitors as candidate agents for the palliative treatment of learning and memory impairments in aging and in neurodegenerative disorders.

Learning-dependent potentiation in the vibrissal motor cortex is closely related to the acquisition of conditioned whisker responses in behaving mice.

The role of the primary motor cortex in the acquisition of new motor skills was evaluated during classical conditioning of vibrissal protraction responses in behaving mice, using a trace paradigm. Conditioned stimulus (CS) presentation elicited a characteristic field potential in the vibrissal motor cortex, which was dependent on the synchronized firing of layer V pyramidal cells. CS-evoked and other event-related potentials were particular cases of a motor cortex oscillatory state related to the increased firing of pyramidal neurons and to vibrissal activities. Along conditioning sessions, but not during pseudoconditioning, CS-evoked field potentials and unitary pyramidal cell responses grew with a time-course similar to the percentage of vibrissal conditioned responses (CRs), and correlated significantly with CR parameters. High-frequency stimulation of barrel cortex afferents to the vibrissal motor cortex mimicked CS-related potentials growth, suggesting that the latter process was due to a learning-dependent potentiation of cortico-cortical synaptic inputs. This potentiation seemed to enhance the efficiency of cortical commands to whisker-pad intrinsic muscles, enabling the generation of acquired motor responses.

Cholinergic septo-hippocampal innervation is required for trace eyeblink classical conditioning.

We studied the effects of a selective lesion in rats, with 192-IgG-saporin, of the cholinergic neurons located in the medial septum/diagonal band (MSDB) complex on the acquisition of classical and instrumental conditioning paradigms. The MSDB lesion induced a marked deficit in the acquisition, but not in the retrieval, of eyeblink classical conditioning using a trace paradigm. Such a deficit was task-selective, as lesioned rats were able to acquire a fixed-interval operant conditioning as controls, and was not due to nonspecific motor alterations, because spontaneous locomotion and blink reflexes were not disturbed by the MSDB lesion. The deficit in the acquisition of a trace eyeblink classical conditioning was reverted by the systemic administration of carbachol, a nonselective cholinergic muscarinic agonist, but not by lobeline, a nicotinic agonist. These results suggest a key role of muscarinic denervation on the acquisition of new motor abilities using trace classical conditioning procedures. It might also be suggested that muscarinic agents would be useful for the amelioration of some associative learning deficits observed at early stages in patients with Alzheimer's disease.

Classical conditioning of eyelid and mystacial vibrissae responses in conscious mice.

The murine vibrissae sensorimotor system has been scrutinized as a target of motor learning through trace classical conditioning. Conditioned eyelid responses were acquired by using weak electrical whisker-pad stimulation as conditioned stimulus (CS) and strong electrical periorbital stimulation as unconditioned stimulus (US). In addition, conditioned vibrissal protraction was obtained pairing either weak electrical whisker-pad stimulation or a tone as CS, with a strong electric shock delivered in the whisker-pad as US. This finding suggests that evolutionary pressure has selected a sensorimotor system capable of constructing conditioned responses on the basis of temporal relationships of stimuli, independently of any putative functional purpose.

Two related forms of memory in the crab Chasmagnathus are differentially affected by NMDA receptor antagonists.

A visual danger stimulus (VDS) elicits an escape response in the crab Chasmagnathus that declines after a few iterative presentations. Long-lasting retention of such decrement, termed context-signal memory (CSM), is mediated by an association between danger stimulus and environmental cues, cycloheximide sensitive, correlated with PKA activity and NFkappa-B activation, positively modulated by angiotensins, and selectively regulated by a muscarinic-cholinergic mechanism. The present research was aimed at studying the possible involvement of NMDA-like receptors in CSM, given the role attributed to these receptors in vertebrate memory and their occurrence in invertebrates including crustaceans. Vertebrate antagonists (+/-)-2-amino-5-phosphonopentanoic acid (AP5) and (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801) were used. Memory retention impairment was shown with MK-801 10(-3) M (1 microg/g) injected immediately before training or after training, or delayed 1 or 4 h, but not 6 h, posttraining. An AP5 10(-3) M dose (0.6 microg/g) impairs retention when given before but not after training. Neither antagonist produced retrieval deficit. A memory process similar to CSM but nonassociative in nature and induced by massed training (termed signal memory, SM), proved entirely insensitive to AP5 or MK-801, confirming the view that distinct mechanisms subserve these different types of memory in the crab.