Ignacio González-Burgos

Serotonin/dopamine interaction in memory formation

Both serotonin (5-HT) and dopamine (DA) neurotransmitters play a key role in modulating synaptic transmission in the central nervous system. Such 5-HT- and DA-mediated modulatory activity has been shown to influence a wide variety of cerebral functions, both of an instrumental and cognitive nature. Some brain regions strongly involved in cognition such as the prefrontal cortex, hippocampal formation and corpus striatum, are densely innervated by serotonergic and dopaminergic afferents proceeding from the raphe complex and the mesocorticolimbic or nigrostriatal systems, respectively. Learning and memory are strongly modulated by 5-HT and DA neurotransmitter activity, and in some cases they interact interdependently to sustain the psychobiological organization of these cognitive processes. Learning and memory, at least in part, depend on short- or long-lasting synaptic modifications, mainly occurring at dendritic spines. Indeed, the modulatory influence of 5-HT and DA at the synaptic level may affect the codification of mnemonic information on such spines. In fact, several experimental models of neurotransmitter activity have identified a close association between a 5-HT-DA imbalance and cytoarchitectonic changes underlying learning and memory impairment.

Place-learning, but not cue-learning training, modifies the hippocampal theta rhythm in rats

Hippocampal theta activity accompanies behaviours such as running, swimming, head movements and spatially orientated responses in the rat. However, whether a relationship between this activity and information processing exists remains unclear. As place-learning depends on hippocampal integrity, whereas cue-learning does not, the hippocampal theta activity underlying each test was evaluated. Local CA1 hippocampal electroencephalograms (EEGs) were recorded over 6 days in a Morris maze for one place-learning test group of rats (n=10) and one cue-learning test group (n=8). The EEG of the corresponding test was taken during waking immobile, searching, and on-platform stages. The relative power (RP) values of EEGs were divided into 4-6.5Hz, 6.5-9.5Hz, and 9.5-12Hz frequency sub-bands. The place-learning training produced a separation of 4-6.5Hz and 6.5-9.5Hz sub-bands in searching and platform stages and a higher activity on the 6.5-9.5Hz sub-band during searching, compared with the basal record, while the cue-learning group did not show differences related either to the training or behavioural stage. As the motor activity and swimming velocity was similar in both groups, the results strongly suggest that the changes observed in theta activity reflect information processing by the hippocampus.

Prenatal and postnatal exposure to ethanol induces changes in the shape of the dendritic spines from hippocampal CA1 pyramidal neurons of the rat.

20%-ethanol was provided to adult female rats since a pregestational stage until weaning of the pups, and percentage proportion of thin, mushroom-shaped, stubby, or wide spines from the apical dendrite of hippocampal CA1 pyramidal cells, was counted at 15, 21, 40, and 90 days of age. By-kind-of-spine analysis revealed higher fluctuation of experimental spines, and less percentage of thin spines was observed in the ethanol-intoxicated rats concomitantly with a higher proportion of stubby or wide spines; through development. Because thin spines may propagate the synaptic potentials more efficiently than stubby or wide spines, this findings suggest that the electrical excitability and thereafter the firing pattern of those cells may be altered, due to the toxic effects of chronic ethanol ingestion.

Golgi Method without Osmium Tetroxide for the Study of the Central Nervous System

A variant Golgi technique was developed that consisted of substituting osmium tetroxide with formaldehyde as the initial fixative in intracardiac perfusion, along with the addition of glacial acetic acid to the chromating fluid. This procedure avoids disposal of dangerous waste substances into the environment. Other advantages include 1) reduction of cost, danger to lab workers, and risk of disruption of the tissue slices during their handling by eliminating the osmium tetroxide, 2) clear tissue background, 3) greater quantity of impregnated neurons than in the classical procedure, with distinct morphological details easily identified even in gross sections and 4) reduction in processing time.

Prefrontocortical serotonin depletion results in plastic changes of prefrontocortical pyramidal neurons, underlying a greater efficiency of short-term memory

The prefrontal cortex activity is involved in organizing the short-term memory. Although the involvement of serotonin for an appropriate performance in learning and memory tests is well known, its role is still unclear; as is the cellular basis of short-term memory behavioral performance. Sprague-Dawley rats were stereotactically injected with 1 microg/microl of 5, 7-dihydroxitryptamine to cause a lesion to the dorsal raphe nucleus. Sham-operated or intact rats were also studied as control groups. Before surgery and 20 days post-operatively, each animal was placed in the Biel maze for five consecutive trials. In the pre-treatment test, all three groups decreased significantly the number of errors beginning with the fourth trial. The same occurred in the post-treatment test, except for the experimental group, whose animals committed less errors beginning with the second trial. After behavioral testing, the dorsomedial prefrontal cerebral cortex was dissected out, and the Golgi study of the third-layer pyramidal neurons revealed that the length of both the apical and the basilar dendrites was smaller than that of controls, and that the apical and oblique dendrites had a greater spine density. A major proportion of thin spines was also seen on the basilar and oblique dendrites, and more stubby spines were seen on the apical dendrite. Serotonin depletion in the prefrontal cerebral cortex resulted in cytoarchitectural alterations of the prefrontocortical pyramidal neurons, which may be underlying partially the greater efficiency observed in the short-term memory behavioral performance.

Increase of the hippocampal theta activity in the Morris water maze reflects learning rather than motor activity

The change in the percentage of rat hippocampal high-frequency theta activity from being immobile and awake to swimming behaviour was calculated for three groups of rats, trained in either place learning, cue learning or egocentric learning in the Morris water maze. The place-learning-trained rats showed an increase in the percentage of theta activity, along with a significant reduction in escape latency over the last 3 days of training. No changes were observed in the other two groups. Because the motor activity displayed by the three groups of rats was similar, we suggest that the increase in the percentage of theta activity concomitant with place-learning training could be related to the processing of information by the hippocampus, rather than to the displayed motor activity.

Long-term morphological and functional evaluation of the neuroprotective effects of post-ischemic treatment with melatonin in rats

Abstract:  Consensus on neuroprotection has pointed out the relevance of the long‐term morphological and functional evaluation of the effectiveness of putative neuroprotective procedures. In the present study, place learning (Morris water maze) and working memory (eight‐arm Olton radial maze) were evaluated in adult male rats 90 days after 15 min of global cerebral ischemia (four‐vessel occlusion) followed by continuous i.v. infusion (10 mg/kg/hr) of melatonin (Isch + Mel) or vehicle (Isch + Veh) for 6 hr, and the pyramidal neuron population of the cornus Ammoni (CA) of the hippocampus and layers III and V of the medial prefrontal cortex was assessed at the end of the behavioral testing period (120 days after ischemia). Impairment of place learning, a significant delay in working memory acquisition, and a significant loss of pyramidal neurons in the Ammons horn (CA1: 23%, CA2: 52% CA3: 73%, hilus: 64% remaining neurons), were observed in the Isch + Veh group. By contrast, a similar performance of the Isch + Mel group to that in the Intact and Sham groups and better than that of the Isch + Veh group, besides a significant reduction of pyramidal neuron loss in the CA subfields (CA1: 79%, CA2: 88% CA3: 86%, hilus: 72% remaining neurons), documented that melatonin treatment led to a long‐term preservation of both the neural substrate, and the capability for integration of spatial learning and memory, mainly dependent on a normal hippocampal functioning. Overall the results emphasize the efficacy of melatonin in counteracting the pathophysiological processes induced by ischemia, by exerting its actions during a short but critical period early after the ischemic episode.

Spine-type densities of hippocampal CA1 neurons vary in proestrus and estrus rats

Spines-mediated synaptic activity has been associated to learning ability. Dendritic spines from hippocampal CA1 pyramidal neurons of proestrus rats have been reported to be more numerous than in estrus animals, but some behavioral studies have reported a better performance during the estrus stage of the estrous cycle. Because spine shape has been shown to be strongly related to the post-synaptic processing of information, a quantitative morphological study related to the proportional density of each type of spine, was conducted in Golgi material of hippocampal CA1 pyramidal cells of proestrus and estrus rats. After three regular estrous cycles had been asserted, seven Sprague-Dawley female adult rats in proestrus and eight in estrus, were used. Mushroom-shaped spines from hippocampal pyramidal cells predominated in proestrous rats, being 15.1% more numerous in this stage than in estrus; while thin spines were the predominant type of spine in estrous animals, being 15.5% more abundant in estrus than in proestrus. The predominance of the mushroom-shaped or thin spines in the hippocampal CA1 pyramidal neurons during the respective stages of the rat estrous cycle, could be related to the organization of the hippocampal activity-dependent mnemonic information.

Long-term study of dendritic spines from hippocampal CA1 pyramidal cells, after neuroprotective melatonin treatment following global cerebral ischemia in rats

Melatonin reduces pyramidal neuronal death in the hippocampus and prevents the impairment of place learning and memory in the Morris water maze, otherwise occurring following global cerebral ischemia. The cytoarchitectonic characteristics of the hippocampal CA1 remaining pyramidal neurons in brains of rats submitted 120 days earlier to acute global cerebral ischemia (15-min four vessel occlusion, and melatonin 10mg/(kg h 6h), i.v. or vehicle administration) were compared to those of intact control rats in order to gain information concerning the neural substrate underlying preservation of hippocampal functioning. Hippocampi were processed according to a modification of the Golgi method. Dendritic bifurcations from pyramidal neurons in both the oriens-alveus and the striatum radiatum; as well as spine density and proportions of thin, stubby, mushroom-shaped, wide, ramified, and double spines in a 50 microm length segment of an oblique dendrite branching from the apical dendrite of the hippocampal CA1 remaining pyramidal neurons were evaluated. No impregnated CA1 pyramidal neurons were found in the ischemic-vehicle-treated rats. CA1 pyramidal neurons from ischemic-melatonin-treated rats showed stick-like and less ramified dendrites than those seen in intact control neurons. In addition, lesser density of spines, lower proportional density of thin spines, and higher proportional density of mushroom spines were counted in ischemic-melatonin-treated animals than those in the sinuously branched dendrites of the intact control group. These cytoarchitectural arrangements seem to be compatible with place learning and memory functions long after ischemia and melatonin neuroprotection.

Neonatal exposure to monosodium L-glutamate induces loss of neurons and cytoarchitectural alterations in hippocampal CA1 pyramidal neurons of adult rats

Glutamatergic post-synaptic receptors are closely related to the known excitotoxic effects of high doses of L-glutamate. Several behavioral abnormalities, glial reaction, and an increase of expression of the NMDA receptor sub-units have been observed in the rat hippocampus after early monosodium glutamate exposure. Thus, a quantitative morphological study was carried out to determine the effects of early exposure to monosodium glutamate on post-synaptic structures that mediate glutamate excitatory neurotransmission in the hippocampal CA1 field. Four milligrams per gram body weight of monosodium glutamate was subcutaneously injected into neonatal Wistar rats, at 1, 3, 5, and 7 days. Cell loss and several cytoarchitectonic parameters were evaluated in pyramidal cells from the hippocampal CA1 field in the treated rats at 60 days of age. An untreated group of rats were used as controls. Cell number in the hippocampus of experimental rats was 11.5% less than that in control animals. In addition, both dendritic arborization and dendritic spine density were adversely affected, and thin and mushroom-shaped spines became proportionally more numerous, while the opposite occurred to stubby spines. These results strongly suggest the occurrence of cell death and also show some cytoarchitectural modifications in the surviving neurons. These could lead to functional alterations in the hippocampal integrative activity, due to an early cytoexcitotoxic effect of monosodium glutamate.