Miguel Cervantes

Neuroprotective effects of progesterone on damage elicited by acute global cerebral ischemia in neurons of the caudate nucleus.

BACKGROUND In addition to the hippocampus, the dorsolateral caudate nucleus (CN) and the pars reticularis of the substantia nigra (SNr) are among the most vulnerable brain areas to ischemia. A possible association of the neuronal injury in these two subcortical nuclei has been proposed, the primary damage affecting the CN GABAergic neurons innervating the SNr, and secondarily the SNr neurons as a result of an imbalance of GABAergic and glutamatergic input to the SNr. Progesterone (P(4)) exerts a GABAergic action on the central nervous system (CNS) and is known to protect neurons in the cat hippocampus from the damaging effect of acute global cerebral ischemia (AGCI). The effects of AGCI on the neuronal populations of the CN and SNr, in addition to the possible neuroprotective effects of P(4), were assessed in cats in the present study. METHODS Ovariectomized adult cats were treated subcutaneously (s.c.) with either P(4) (10 mg/kg/day) or corn oil during the 7 days before and 7 days after being subjected to a period of AGCI by 15 min of cardiorespiratory arrest followed by 4 min of reanimation. After 14 days of survival, animals were sacrificed and their brains perfused in situ with phosphate-buffered 10% formaldehyde for histologic examination. RESULTS ACGI resulted in an intense glial reaction in the CN and a significant loss (43%) of medium-sized neurons of the CN, but no difference was found in the densities of SNr neurons between controls and ischemic oil- and P(4)-treated cats. Progesterone treatment completely prevented CN neuronal loss. CONCLUSIONS The overall results point to the higher vulnerability of CN neurons to ischemia as compared to neurons in the SNr and show the protective effects of P(4) upon CN neuronal damage after ischemia.

Post-ischemic administration of progesterone in rats exerts neuroprotective effects on the hippocampus.

Progesterone is neuroprotective in models of focal or global ischemia when treatment starts either before the insult or at the onset of reperfusion. In these cases the steroid may act during the occurrence of the early pathophysiological events triggered by ischemia or reperfusion. As opposed to this condition, the aim of the present study was to assess the effect of delayed, post-injury administration of progesterone on the preservation of pyramidal neurons of the hippocampus of rats 21 days after been exposed to global ischemia by the four vessel occlusion model. Progesterone (8 mg/kg, i.v.) or its vehicle, were administered at 20 min, 2, 6, and 24h after the end of ischemia. At histological examination, brains of the ischemic vehicle-treated rats showed a severe reduction of the population of pyramidal neurons in the CA1 and CA2 subfields (12% and 29% remaining neurons, respectively), and a less severe neuronal loss in the CA3 and CA4 subfields of the hippocampus (68% and 63% remaining neurons, respectively), as compared to rats exposed to sham procedures. They also showed a two-fold enlargement of the lateral ventricles and 33% shrinkage of the cerebral cortex as compared to the sham group. Progesterone treatment resulted in a significant preservation of pyramidal neurons in CA1 and CA2 (40% and 62% remaining neurons), with no ventricular dilation and only a mild (12%) cortical shrinkage. Results suggest that progesterone is able to interfere with some late pathophysiological mechanisms leading both to selective neuronal damage in the hippocampal CA1 and CA2 subfields, and to shrinkage of the cerebral cortex.

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.

Neuroprotective Effect of Melatonin on Brain Damage Induced by Acute Global Cerebral Ischemia in Cats

BACKGROUND Melatonin has been proposed as a neuroprotective agent on the basis of its ability to function as a free radical scavenger, provided that lipoperoxidation and other free radical damage induced by reactive oxygen species resulting from cerebral ischemia are relevant pathophysiologic processes of ischemic neuronal damage. METHODS The effects of melatonin or vehicle on neurologic deficit scores (Todd scale; maximal deficit score = 100, daily during 7 days after the ischemic episode) and neuronal population of hippocampal CA1-CA4 fields (cresyl violet stain, at the eighth day after the ischemic episode) were evaluated in adult male cats subjected to a 15-min period of acute global cerebral ischemia induced by cardiorespiratory arrest, and in cats subjected to a sham procedure. Continuous intravenous (iv) administration of either melatonin 10 mg/kg/h in 10% ethanol in saline or the vehicle alone (3 mL/kg/h) for 6 h starting 30 min after the end of the period of global cerebral ischemia was used as treatment. RESULTS Global cerebral ischemia resulted in a severe loss of neurons in hippocampal CA1-CA4 fields (9, 13, 30 and 28% remaining neurons, respectively) of ischemic vehicle-treated cats in comparison with sham cats (100%). By contrast, remaining neurons in these regions were between 81 and 100% in the ischemic melatonin-treated cats, values that are nonsignificantly different as compared with sham cats. Values of remaining neurons in CA1-CA4 fields in ischemic melatonin-treated cats were significantly higher than those in ischemic vehicle-treated cats. Neurologic deficit scores in ischemic vehicle-treated cats (42-77 at day 1, 6-39 at day 7) were significantly higher than those in ischemic melatonin-treated cats (16-38 at day 1, 0-6 at day 7) on the days after the ischemic episode. CONCLUSIONS Overall, the results support the neuroprotective effect of melatonin against the neuronal cerebral damage induced by acute global cerebral ischemia.

Neuroprotective effects of progesterone and allopregnanolone on long-term cognitive outcome after global cerebral ischemia

PURPOSE To assess the longterm neuroprotective effects of progesterone (P₄) and allopregnanolone (ALLO) on functional and morphological parameters of the integrity of the hippocampus, after global cerebral ischemia. METHODS Adult male Sprague-Dawley rats were subjected to a transient severe (20 min) forebrain ischemia (Isch) episode and treated with P₄ or ALLO (8 mg/kg i.v.) or its vehicle, at 20 min, 2, 6, 24, 48 and 72 h after ischemia. Rats subjected to Sham procedures, and intact rats were included as nonischemic controls. Three months after ischemia, both the functional (spatial learning and memory, and reference and working memory), and the morphological integrity (dimensions of the hippocampal formation, thickness of the CA1 subfield, and pyramidal neuron population) of the hippocampus and the medial prefrontal cortex(mPFC) were determined. RESULTS Treatment with P₄ or ALLO significantly reduced the impairment in spatial learning and memory, as well as in reference and working memory, and prevented the narrowing of the hippocampus, otherwise induced by ischemia. This better performance of P₄ and ALLO treated rats than vehicle (Veh) treated rats, occurred in spite of a loss of pyramidal neurons in the CA1, CA2,CA3 and hilus subfields of the Ammons horn (remaining neurons: Isch+Veh: 21.0, 35.6, 44.1, and 40.3%; Isch+P₄: 19.9, 32.2,41.1, and 32.5%; Isch+ALLO: 25.5, 62.0, 73.7, and 56.7%), and nonsignificant changes in the mPFC, as compared to the Intact group (100%). CONCLUSIONS Performance of P₄ or ALLO treated rats in learning and memory tests suggests that these steroids promoted neural conditions accounting for adequate functioning long after ischemia, in spite of the loss of hippocampal pyramidal neurons.

Characteristic frequency bands of the cortico-frontal EEG during the sexual interaction of the male rat as a result of factorial analysis

The electrocorticogram (ECoG) from the prefrontal cortex was simultaneously recorded with the accelerometric signals of pelvic thrusting performed by male rats during sexual behavior. The changes in the prefrontal ECoG were precisely correlated in time with well defined elements of male rat copulation. Principal component analysis allowed to identify three distinct bands of frequencies in the frontal ECoG: the absolute power (AP) of the 4-16 Hz band was increased in the 500-ms periods before, during, and after the execution of pelvic thrusting in mount, intromission and ejaculation responses; the AP of the 18-24 Hz band was selectively increased during the execution of pelvic thrusting at the three copulatory responses, whereas the AP of the 26-32 Hz band was increased only during the pelvic moments of mount and intromission responses. These results show that the electroencephalographic activity of the prefrontal cortex of the male rat is related to the performance of sexual behavior, supporting the concept that this cortical area is involved in the organization of sequential behaviors, as sexual behavior.

Subcortical multiple unit activity changes during rat male sexual behavior

Multiple unit activity (MUA) was recorded from the ventral tegmental area (VTA) and mesencephalic locomotor region (MLR) during copulatory behavior of freely moving male rats. Simultaneous accelerometric recordings of the copulatory pelvic thrusting performed by the male rat were taken to precisely correlate in time the changes in MUA with well defined elements of copulation. The baseline MUA firing rates recorded in the quiet-alert condition in the VTA and in the MLR were significantly increased during pursuit of the female by the male; significantly higher MUA firing rates were found in the VTA at the 500 ms periods before and during the execution of pelvic thrusting in mount, intromission, and ejaculation responses as compared to the baseline, and returned to this value when these responses ended. The maximum MUA firing rate in the MLR was obtained during the execution of pelvic thrusting in each copulatory response, and it remained significantly elevated, as compared to the baseline, after thrusting and at the postintromission and postejaculatory genital grooming, then decreasing to basal values at the initial part of the postejaculatory interval. The fact that the highest changes in MUA were related to pelvic thrusting suggests a major participation of both structures in the execution of motor copulatory responses.

Analysis of various factors involved in EEG synchronization during milk drinking in the cat

Electroencephalographic (EEG) and multineuronal activity (MUA) from several brain structures were recorded in 11 adult cats during milk drinking. Milk drinking elicited parietal and hippocampal EEG synchronization and inhibited multiunit discharge in several brain regions. The proportion of EEG synchronization varied considerably between animals, but remained more or less constant for each cat after several testing days. Neither modification of milk concentration nor fasting influenced the proportions of EEG synchronization during milk drinking. Adaptation of the animals to the experimental environment facilitated the appearance of EEG synchronization during milk drinking. Chlorpromazine (3 mg/kg) significantly increased the proportion of EEG synchronization observed during milk drinking. By contrast, displacement of the head, neck or postural body adjustments during milk drinking blocked EEG synchronization. Amphetamine also had a suppressive effect on the EEG parietal and hippocampal synchronic activity observed during milk drinking. Our results suggest that development of parietal EEG synchronization in response to pleasant stimulation is a complex phenomenon requiring a relaxed condition of the animal, including its adaptation to the experimental environment and relative immobility with the concomitant diminution of afferent inflow from some of the muscles involved in locomotion.