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Neuroprotective and neurotoxic effects of estrogens

The ovarian hormone 17beta-estradiol (E2) is neuroprotective in animal models of neurodegenerative diseases. Some studies suggest that the neuroprotective effects of 17beta-estradiol are a consequence of its antioxidant activity that depend on the hydroxyl group in the C3 position of the A ring. As in other tissues, 17beta-estradiol is metabolized in the brain to 2-hydroxyestradiol (2OHE2) and 2-methoxyestradiol (2MEOHE2). These two molecules present the hydroxyl group in the A ring and have a higher antioxidant activity than 17beta-estradiol. To test the hypothesis that conversion to 2-hydroxyestradiol and 2-methoxyestradiol may mediate neuroprotective actions of 17beta-estradiol in vivo, we have assessed whether these molecules protect hilar hippocampal neurons from kainic acid toxicity. Ovariectomized Wistar rats received an i.p. injection of 1, 10 or 100 microg 17beta-estradiol, 2-hydroxyestradiol or 2-methoxyestradiol followed by an i.p. injection of kainic acid (7 mg/kg) or vehicle. Treatment with kainic acid resulted in a significant loss of hilar neurons. Only the highest dose tested of 17beta-estradiol (100 microg/rat) prevented kainic acid-induced neuronal loss. 2-Hydroxyestradiol and 2-methoxyestradiol did not protect hilar neurons from kainic acid, suggesting that the mechanism of neuroprotection by 17beta-estradiol in vivo is not mediated by its metabolism to catecholestrogens or methoxycatecholestrogens. Furthermore, 2-methoxyestradiol (100 microg/rat), by itself, resulted in a significant neuronal loss in the hilus that was detected 96 h after the treatment with the steroid. This finding suggests that endogenous metabolism of 17beta-estradiol to 2-methoxyestradiol may counterbalance the neuroprotective effects of the hormone.

Inhibitory effect of buspirone and diazepam, but not of 8-OH-DPAT, on maternal behavior and aggression

The action of diazepam (0.0, 1.0, and 2.0 mg/kg) and the serotonergic compounds buspirone (0.0, 2.5, and 5.0 mg/kg) and 8-OH-DPAT (0.0, 0.1, and 1.0 mg/kg) on maternal behavior and aggression were studied. An activity test was made after these treatments to control for unspecific actions due to motor impairment. Diazepam and buspirone dose-dependently inhibited the expression of maternal aggression and the active components of maternal behavior such as retrieving and nest building. 8-OH-DPAT did not affect these behaviors. 8-OH-DPAT (1.0 mg/kg) provoked the serotonergic syndrome and hypothermia; however, ovariectomized animals showed more signs of the syndrome and a decrease in body temperature after 8-OH-DPAT than lactating rats. Buspirone, but not the other anxiolytics, reduced motor activity. The role of drugs acting at the serotonergic, dopaminergic, and GABA-benzodiazepine systems in the control of maternal behavior and aggression is discussed.

Alterations in GABAergic function following forced swimming stress.

Forced swimming induces alterations in the GABA brain concentration and could change the sensitivity of the GABA/benzodiazepine receptor-chloride ionophore complex to benzodiazepines. This change in sensitivity could be explained by the allopregnanolone release that takes place during stress. The current study was carried out to determine whether forced swimming is able to modify the anti-anxiety effect of diazepam and to explore the possible relation of this change to allopregnanolone, the GABA concentration or/and the GABA/benzodiazepine receptor density. Unstressed and stressed mice, injected with the vehicle or diazepam, were evaluated in the exploratory behavior test. Diazepam induced clear anxiolytic actions at all doses in unstressed animals, but such an effect was not observed in stressed animals. The injection of allopregnanolone 24 h before the anxiety test blocked the effect of this benzodiazepine. Forced swimming decreased GABA concentrations in the hippocampus and the thalamus-hypothalamus region, besides decreasing the [(3)H]flunitrazepam labeling in both the hypothalamus and amygdala. These results show that forced swimming abolishes the anti-anxiety effect of diazepam.

Influence of forced swimming stress on 5-HT1A receptors and serotonin levels in mouse brain

Several stressful factors are able to modify 5-HT1A receptors; for example, different schemes of forced swimming-induced stress (FS) produce a variety of changes in synthesis as well as in 5-HT1A binding in the brain. In addition, it is known that the concentration of 5-HT in the brain is modified as a consequence of acute stressing. The main purpose of this study was to characterize the influence of 15 min of FS on 5-HT levels and on 5-HT1A receptor density in specific brain areas. Mice stressed 24 h before were sacrificed and their brains processed by means of a quantitative autoradiography technique. The following areas were studied: dorsal raphe nucleus (DRN); median raphe nucleus (MRN); thalamus; hypothalamus; amygdala, and hippocampus. 5-HT and 5-hydroxyindolacetic acid (5-HIAA) concentrations in the brainstem, thalamus-hypothalamus, and hippocampus of stressed (ST) mice were analyzed 24 h after stressing by high performance liquid chromatography (HPLC) with fluorometric detection. All data were compared with corresponding unstressed (UST) controls. A significant decrease in 5-HT1A receptor density in DRN, MRN, and hippocampus, accompanied by an increase in labeling of 5-HT1A receptor in thalamus, hypothalamus, and amygdala was observed in ST animals. FS induced a decrease in the 5-HT concentration in the thalamus-hypothalamus, accompanied by an increase in hippocampus areas without affecting 5-HT concentration in the brainstem. Additionally, 5-HIAA/5-HT ratio in the thalamus-hypothalamus area was increased. This study showed that stress alters both 5-HT concentration and 5-HT1A receptors in a region-specific manner.

Long-term ovariectomy modulates the antidepressant-like action of estrogens, but not of antidepressants

Controversial results related to effectiveness of estrogen replacement therapy (ERT) to alleviate depression are frequently reported. The discrepancies could be related to (a) time when ERT is initiated after the beginning of menopause and/or (b) type of estrogen used. Furthermore, estrogens modulate the antidepressant effect of different compounds; therefore, the effectiveness of antidepressant drugs could also depend on the menopausal status. The aim of the present study was to analyze whether the time after estrogen decline can influence antidepressant-like effects of two estrogens and/or two antidepressants. Thus, the antidepressant-like actions of 17β-estradiol (E2), 17α-ethynyl-estradiol (EE2), fluoxetine (FLX) and desipramine (DMI) were studied at different periods (1, 3 and 12 weeks) after ovariectomy (OVX), using the forced swimming test (FST). Results showed that OVX increased depressive-like behavior only 1 week after OVX. The antidepressant-like actions of E2, but not those of EE2, were cancelled 12 weeks after OVX. Conversely, antidepressant-like actions of FLX and DMI were observed at 1, 3 and 12 weeks after OVX. In conclusion, while the antidepressant-like effects of estrogens depended on the time at which treatment is initiated after OVX as well as on the estrogenic compound used, antidepressant-like effects of FLX and DMI were not blocked by OVX.

Different expression of α and β mitochondrial estrogen receptors in the aging rat brain: Interaction with respiratory complex V

Recent evidence suggests that hormonal effects on mitochondria could be mediated by mitochondrial estrogen receptors (mtERs). These receptors are new candidates for the beneficial estrogenic effects on mitochondria in different physiological conditions. The aim of this investigation was to study mtER expression during brain aging. We analyzed mtERalpha and mtERbeta expression in cortical, hippocampal and hypothalamic mitochondria of young adult (3months) and aged (18 months) female Wistar rats by Western blot. In addition, we explored the interaction of mtERbeta with respiratory complex V by using coimmunoprecipitation assays. The results show that mtERalpha and mtERbeta are present in young and aged brain mitochondria. We also demonstrate that mtERs are expressed as variants and have a brain region specific distribution. The predominant mtER variants detected were of 61 and 55KDa for mtERalpha and of 63 and 52KDa for mtERbeta. However, we did not observe differences in the mtERalpha or beta content between the two age groups studied. Additionally, we show that mtERbeta interacts with complex V. The overall results demonstrate that there is a differential expression of mtERalpha and mtERbeta variants in different brain areas, indicating that they may participate in different functions in the brain during aging.

Differential Expression of Estrogen Receptors in Two Hippocampal Regions During the Estrous Cycle of the Rat

In the hippocampus, estrogens increase dendritic arborization, long‐term potentiation, neuroprotection, and participate in many functions related with learning, memory, and affective behaviors. The presence of both estrogen receptors alpha (ERα) and beta (ERβ) isoforms has been described in the hippocampus where they play different physiological roles. The aim of this study was to investigate, by using both techniques immunohistochemistry and Western Blot, the expression pattern of ERα and ERβ in the hippocampus of the rat along the estrous cycle. Western blot analysis was used to confirm the specificity of the antibodies used against ERα and ERβ and its relative content in the hippocampus. Results from immunohistochemical studies indicate that ERβ expression increased more than the ERα in CA1 and CA3 regions during all phases of the estrous cycle. ERβ immunoreactivity was mainly located in the nucleus and predominantly expressed in CA1 during estrous and metestrus, and in CA3 during diestrus. ERα was more abundant during estrous in comparison to other phases of the cycle in CA1 region, while it was more abundant during metestrus in CA3. Interestingly, the immunolocalization of ERα subtype was both cytoplasmic and nuclear. The overall results indicate that there is a differential expression, cellular localization, and distribution of both ER subtypes in CA1 and CA3 regions, suggesting different roles for these two receptors in the hippocampus along the estrous cycle. Anat Rec, 2011.

Blockade of the anxiolytic action of 8-OH-DPAT in lactating rats.

The anxiolytic action of 8-OH-DPAT (0.125, 0.5, and 0.75 mg/kg, I.P., -15 min) was evaluated in ovariectomized and 7-day lactating mother rats. This serotonergic anxiolytic produces a clear reduction in cumulative burying behavior and in freezing time (parameters denoting a reduction in anxiety levels) in ovariectomized subjects. However, in 7-day lactating mother rats, 8-OH-DPAT lacked its antianxiety effects. Additionally, as an extra parameter of the animals emotionality the number of defecation bolus was registered. A reduction in this parameter in ovariectomized animals after 8-OH-DPAT (0.5 mg/kg) injection was found. This effect was also blocked in 7-day lactating rats. None of the doses of the compound tested altered the burying behavior latency (a parameter inversely representing the animals reactivity). The general ambulatory behavior of ovariectomized subjects was significantly impaired at the highest dose (0.75 mg/kg) of 8-OH-DPAT. Lower doses did not alter this parameter or the number of times lines are crossed (a parameter denoting exploratory activity). Discussion is focused on the relationship between the activity of the serotonergic system and the secretion of prolactin in this period.

Effects of short-term hormonal replacement on learning and on basal forebrain ChAT and TrkA content in ovariectomized rats.

It has been proposed that sex steroid hormones improve performance in some cognitive tasks by regulating the basal forebrain cholinergic function. However, the molecular basis of such influence still remains unknown. Current study analyzed the performance of ovariectomized rats in an autoshaping learning task after a short-term treatment with 17β-estradiol (E2: 4 and 40μg/kg) and/or progesterone (P4: 4mg/kg). These results were correlated with basal forebrain choline acetyltransferase (ChAT) and TrkA protein content. The high dose of E2 enhanced both acquisition in the autoshaping task and the content of ChAT and TrkA. P4 treatment increased ChAT and TrkA content without affecting performance of rats in the autoshaping learning task. Interestingly, the continuous and simultaneous administration of E2 plus P4 did not significantly modify behavioral and biochemical evaluated parameters. These results address the influence of both E2 and P4 on cholinergic and TrkA activity and suggest that the effects of ovarian hormones on cognitive performance involve basal forebrain cholinergic neurons.

A-ring reduced derivatives of two synthetic progestins induce anxiolytic effects in ovariectomized rats.

The putative anxiolytic action of the synthetic progestins, norethisterone (NET), levonorgestrel (LNG) and their respective 5 alpha-reduced metabolites, (5 alpha-NET), (3 alpha,5 alpha-NET), (3 beta,5 alpha-NET), (5 alpha-LNG), (3 alpha,5 alpha-LNG) and (3 beta,5 alpha-LNG), were studied in the burying behavior paradigm. From these compounds only 3 alpha,5 alpha-NET and 3 beta,5 alpha-LNG reduced burying behavior without modifying other parameters. Burying behavior latency was prolonged after administration of the highest dose (1.0 mg/rat) of NET and 5 alpha-NET. As a positive control 3 alpha,5 alpha-pregnanolone (allopregnanolone) was included at the same doses used for synthetic steroids. This steroid produced a clear dose-dependent reduction in burying behavior without affecting latency. No steroid with anxiolytic properties affected the general ambulatory behavior. All synthetic steroids with anxiolytic activity proved to be less potent than allopregnanolone. Results are discussed on the basis of the chemical structure requirements necessary to induce tranquilizing effects.