Erica do Espirito Santo Hermel

Endurance and Resistance Exercise Training Programs Elicit Specific Effects on Sciatic Nerve Regeneration After Experimental Traumatic Lesion in Rats

Objective. To evaluate the effects of endurance, resistance, and a combination of both types of exercise training on hindlimb motor function recovery and nerve regeneration after experimental sciatic nerve lesion in rats. Methods. Sciatic nerve crush was performed on adult male rats, and after 2 weeks of the nerve lesion, the animals were submitted to endurance, resistance, and a combination of endurance-resistance training programs for 5 weeks. Over the training period, functional recovery was monitored weekly using the Sciatic Functional Index (SFI) and histological and morphometric nerve analyses were used to assess the nerve regeneration at the end of the trainings. Results. The SFI values of the endurance-trained group reached the control values from the first posttraining week and were significantly better than both the resistance-trained group at the first, second, and third posttraining weeks and the concurrent training group at the first posttraining week. At the distal portion of the regenerating sciatic nerve, the endurance-trained group showed a greater degree of the myelinated fiber maturation than the sedentary, resistance-trained, and concurrent training groups. Furthermore, the endurance-trained group showed a smaller percentage area of endoneurial connective tissue and a greater percentage area of myelinated fibers than the sedentary group. Conclusion . These data provide evidence that endurance training improves sciatic nerve regeneration after an experimental traumatic injury and that resistance training or the combination of 2 strategies may delay functional recovery and do not alter sciatic nerve fiber regeneration.

Influence of sex and estrous cycle, but not laterality, on the neuronal somatic volume of the posterodorsal medial amygdala of rats.

The aim of the present study was to measure the cell body volume of neurons from the posterodorsal subnucleus of the medial amygdala (MePD) of adult male (n=5) and diestrus, proestrus and estrus female (n=4-5 in each group) rats to reveal a possible sexual dimorphism, estrous cycle variations and laterality in this morphological parameter. The brains of adult Wistar rats were sectioned (1 microm), stained with 1% toluidine blue and the stereological estimation of neuronal soma volume of both sides of MePD was realized using the Cavalieri method and the technique of point counting. Data were compared by a two-way ANOVA for repeated measures and the least significance difference post hoc test. In the MePD, mean neuronal somatic volume showed a statistical difference among groups (p=0.005), but neither an effect of laterality (p=0.33) nor interactions between groups and laterality (p=0.78) were found. Post hoc test showed that males (mean+/-S.E.M., 2075.67+/-135.79 microm(3)) have larger mean neuronal somatic volume compared to females in proestrus (1503.30+/-44.46 microm(3)) and in estrus (1616.69+/-71.49 microm(3), p<0.05 in both cases), but not in diestrus (1940.78+/-129.68 microm(3), p>0.05). Moreover, diestrus females displayed larger mean neuronal somatic volume than proestrus female rats (p<0.05). It is suggested that neuronal somatic volume is another sexually dimorphic finding in the MePD, for which it is relevant to set apart the different phases of the estrous cycle to reveal the presence of gonadal hormones effects in the rat MePD neurons.

Gonadal hormone regulation of glial fibrillary acidic protein immunoreactivity in the medial amygdala subnuclei across the estrous cycle and in castrated and treated female rats

The medial amygdala (MeA) is a sexually dimorphic area that modulates neuroendocrine and behavioral activities and where gonadal hormones play an important role in neuron-glial and synaptic plasticity. Immunohistochemistry was used to identify the astrocytic marker glial fibrillary acidic protein (GFAP) in the different MeA subnuclei--anterodorsal (MeAD), posterodorsal (MePD) and posteroventral (MePV)--of intact female rats in the different phases of the estrous cycle and in ovariectomized females treated with hormonal substitutive therapy. Data semi-quantified by optical densitometry showed that, in the proestrus phase, the GFAP immunoreactivity (GFAP-ir) was higher when compared to the other phases of the estrous cycle (P < 0.02). GFAP-ir was also higher in the MePD than in the MeAD or in the MePV (P < 0. 02). In ovariectomized females, injections of estradiol alone or estradiol plus progesterone increased GFAP-ir in the MePD and in the MePV (P < 0.001), but not in the MeAD (P > 0.3), when compared to control data. These findings suggest that astrocytic GFAP in the MeA subnuclei can be affected either by physiological levels or by hormonal manipulation of the ovarian steroids, which may contribute to the plasticity of local and integrated functional activities of these brain areas in female rats.

Neuronal somatic volume of posteroventral medial amygdala cells from males and across the estrous cycle of female rats.

The posteroventral medial amygdala (MePV) is a brain area where gonadal hormones have neurotrophic effects in rats. The aim of the present study was to estimate the MePV neuronal somatic volume from males and diestrus, proestrus and estrus female Wistar rats (n=5 in each group) in an attempt to identify a possible sexual dimorphism in this parameter. The effect of laterality was also evaluated. The brains of adult animals were sectioned (1 microm), stained with 1% toluidine blue and serial-section reconstructions of each neuronal cell body were obtained. Images from both left and right MePV were studied and the somatic volume was estimated using the Cavalieri method in combination with the point counting technique. Results were compared according to sex and phase of the estrous cycle using a two-way ANOVA for repeated measures followed by the least significance difference test. Mean neuronal somatic volume showed a statistical difference among groups and the post hoc comparisons revealed that males present higher values than females in proestrus and estrus (p<0.05). On the other hand, neither a laterality effect (p=0.6) nor an interaction between groups and laterality (p=0.4) were found. Our results indicate that cell body volume in the MePV is distinct when comparing males to females in the different phases of the estrous cycle. Through dynamic changes modulated by sex steroids, it is likely that this morphological plasticity within the MePV may be affecting the functioning of local neurons and their integrated roles in neural circuits relevant for neuroendocrine control and reproductive behaviors.

Neonatal handling increases fear and aggression in lactating rats

Neonatal handling reduces fear in male and cycling female rats, but increases maternal aggressive behavior against intruders to the nest area. Present study aimed to analyze the effects of neonatal handling on the maternal aggressive behavior and the activity in the open field with a predator of lactating rats on the 8th and the 18th postpartum days (periods of high and low aggressiveness). As pups, animals were divided into two groups: nonhandled (no neonatal manipulation) and handled (handling for 1 min during the first 10 days after delivery). As adults, females of both groups were impregnated and tested against a male intruder for aggressive behavior and in the open field with a cat inside a wire-meshed cage. Results showed that on the 8th day frequency of aggressive behaviors of handled females was higher than that of the nonhandled ones, but on the 18th day, no significant difference was detected. Surprisingly, in the open field test, handled females showed decreased locomotion and increased freezing on the 8th day compared to the nonhandled ones. The opposite relationship between increased aggressiveness with reduced fear is observed in the nonhandled control females in early and late lactation periods. However, neonatal handling abolishes this relationship. Apparently, the increased aggressiveness in neonatal handled lactating females does not depend on a decrease in fear. Our findings support the hypothesis that long lasting effects of early life stimulation is a dynamic function depending on the behavioral system and the period of life analyzed. Moreover, they caution the relationship between aggressive behavior and fear.

Influence of substitutive ovarian steroids in the nuclear and cell body volumes of neurons in the posterodorsal medial amygdala of adult ovariectomized female rats.

The volumes of the neuronal nucleus and the cell body in the left posterodorsal medial amygdala (MePD) of adult ovariectomized (OVX) female rats submitted to different hormonal therapies were studied here, aiming to reveal possible influence of substitutive sex steroids in these morphological parameters. One week following ovariectomy and at the end of treatments, brains were cut to semi-thin sections (1 microm) and stained with 1% toluidine blue for stereological estimations, carried out using the Cavalieri method and the technique of point counting. Both the volume of the neuronal nucleus and the soma showed a statistically significant difference when comparing the data among OVX females treated with vehicle (V), estradiol (EB) alone, EB plus progesterone (EB+P) or P alone [n=5 rats in each group; one-way ANOVA test, P<0.01 in both cases]. The Tukey test showed that OVX and EB+P treated females had higher mean neuronal nucleus and somatic volumes when compared to V (P<0.01) or EB alone (P<0.01). Also, OVX females treated with P alone showed larger mean neuronal nucleus and somatic volumes when compared to V (P<0.05). These results suggest that the neuronal nucleus and the somatic volumes can be modulated by substitutive ovarian hormones administered to OVX females, for which P can lead to higher results. These findings reveal additional epigenetic actions of the sex steroids in the MePD and new neuronal morphological features in adult female rats.

Neonatal handling and the expression of immunoreactivity to tyrosine hydroxylase in the hypothalamus of adult male rats

Neonatal handling has long-lasting effects on behavior and stress reactivity. The purpose of the present study was to investigate the effect of neonatal handling on the number of dopaminergic neurons in the hypothalamic nuclei of adult male rats as part of a series of studies that could explain the long-lasting effects of neonatal stimulation. Two groups of Wistar rats were studied: nonhandled (pups were left undisturbed, control) and handled (pups were handled for 1 min once a day during the first 10 days of life). At 75-80 days, the males were anesthetized and the brains were processed for immunohistochemistry. An anti-tyrosine hydroxylase antibody and the avidin-biotin-peroxidase method were used. Tyrosine hydroxylase-immunoreactive (TH-IR) neurons were counted bilaterally in the arcuate, paraventricular and periventricular nuclei of the hypothalamus in 30-microm sections at 120-microm intervals. Neonatal handling did not change the number of TH-IR neurons in the arcuate (1021 +/- 206, N = 6; 1020 +/- 150, N = 6; nonhandled and handled, respectively), paraventricular (584 +/- 85, N = 8; 682 +/- 62, N = 9) or periventricular (743 +/- 118, N = 7; 990 +/- 158, N = 7) nuclei of the hypothalamus. The absence of an effect on the number of dopaminergic cells in the hypothalamus indicates that the reduction in the amount of neurons induced by neonatal handling, as shown by other studies, is not a general phenomenon in the brain.

Effect of skilled and unskilled training on nerve regeneration and functional recovery

The most disabling aspect of human peripheral nerve injuries, the majority of which affect the upper limbs, is the loss of skilled hand movements. Activity-induced morphological and electrophysiological remodeling of the neuromuscular junction has been shown to influence nerve repair and functional recovery. In the current study, we determined the effects of two different treatments on the functional and morphological recovery after median and ulnar nerve injury. Adult Wistar male rats weighing 280 to 330 g at the time of surgery (N = 8-10 animals/group) were submitted to nerve crush and 1 week later began a 3-week course of motor rehabilitation involving either “skilled” (reaching for small food pellets) or “unskilled” (walking on a motorized treadmill) training. During this period, functional recovery was monitored weekly using staircase and cylinder tests. Histological and morphometric nerve analyses were used to assess nerve regeneration at the end of treatment. The functional evaluation demonstrated benefits of both tasks, but found no difference between them (P > 0.05). The unskilled training, however, induced a greater degree of nerve regeneration as evidenced by histological measurement (P < 0.05). These data provide evidence that both of the forelimb training tasks used in this study can accelerate functional recovery following brachial plexus injury.