Lucía Martínez-Mota

Anxiolytic-like actions of testosterone in the burying behavior test: role of androgen and GABA-benzodiazepine receptors

The first objective of the present study was to explore if several androgens, including testosterone propionate (TP) and its reduced metabolites, 5alpha-androstan-3alpha, 17beta-diol (3alpha-androstanediol) and 5alpha-androstan-3alpha-ol-17-one (androsterone), produce an anxiolytic-like effect in the burying behavior paradigm. Additionally, the possible participation of androgen or GABA(A)-benzodiazepine receptors in the anxiolytic-like effect of TP was analyzed. Orchidectomized male rats were treated with four injections of TP (0.25, 0.50, 1.0 mg/rat), 3alpha-androstanediol or androsterone (0.5 and 1.0 mg/rat), and the cumulative burying (denoting anxiety levels) and burying behavior latency (reflecting reactivity) evaluated. Besides, a single injection (0.5 mg/rat, -30 m) of each androgen was also tested in castrated rats. Repeated treatment with TP (0.5 and 1.0 mg/rat), but not a single injection of this androgen, produced an anxiolytic-like effect without changes on reactivity. Neither repeated- nor single-treatment with 3alpha-androstanediol nor androsterone produced a reduction of anxiety-like behavior. For the experiment studying the TP mechanism of action, this androgen (0.5 mg/rat, four injections) was combined with an antagonist for androgen receptors, flutamide (50 mg/kg, SC, 7 injections), or with a benzodiazepine antagonist, flumazenil (15 mg/kg, IP, -30 m). Flutamide, but not flumazenil, blocked the anxiolytic-like effects produced by TP. Results are discussed on the basis of interaction of these steroids with androgen receptors or GABA(A)-benzodiazepine receptors.

Desipramine restricts estral cycle oscillations in swimming.

1. Desipramine (DMI) is a tricyclic antidepressant which reduces the immobility in rats forced to swim; however, it is unknown whether estral cycle phases impinge on DMI actions on immobility in daily swimming tests during several weeks. 2. In female wistar rats, vaginal smears taken before testing defined four estral phases. Afterwards, the authors assessed the latency for the first period of immobility in five-min forced swim tests practiced on 21-day DMI (DMI group), 21-day washout saline given after a 21-day DMI treatment (washout-saline group), or non-treated rats (control group). 3. We observed a longer latency for the first period of immobility in proestrus-estrus from the control and washout-saline groups. The 21-day treatment with DMI (2.1 mg/kg i.p., once a day) significantly (p < 0.001) increased the latency by about 160% from control regardless of the estral cycle phase. 4. It is concluded that proestrus-estrus relates to increased struggling behavior. DMI enhances struggling behavior independently of hormonal state.

Lateral septal neuronal firing rate increases during proestrus-estrus in the rat

Neuronal activity of the lateral septal nucleus (LSN) is related to motivational and hedonic behavior. Even though some changes in mood and anxiety during proestrus and pregnancy have been reported, the possible changes in the neuronal activity of the LSN through the phases of the estrous cycle are unknown. In the present study we explored the neuronal activity from the LSN using glass micropipettes (NaCl 1 M, and Evans blue 2.5%; 3-8 Mohms in 30 urethane (1 g/kg) anesthetized Wistar rats. Analysis of data included a total of 88 single-unit extracellular recordings taken from the LSN during proestrus (n = 22), estrus (n = 23), diestrus (n = 22), and metestrus (n = 21). The highest values of firing rate were found in proestrus, and the lowest in metestrus, F(3,84) = 3.78, p < 0.01. During estrous cycles, in the phase characterized by high plasma levels of estradiol and progesterone, i.e., proestrus-estrus, the neurons from the dorsal aspect of the LSN fired at significantly (p < 0.05) higher frequencies, shorter first-order intervals and a lower coefficient of variation than those in the phase characterized by lower levels of estradiol and progesterone (metestrus-diestrus). In another group of rats (n = 12), immobility in the forced-swim test was assessed. Consistently, a longer latency (p < 0.05) for the first period of immobility and a nonsignificant trend to a lowered total time in immobility were found in proestrus and estrus. It is concluded that the higher firing rate in neurons from the dorsal aspect of the LSN during proestrus-estrus, may be associated with an increased motivation to escape from a stressful situation.

Interaction of desipramine with steroid hormones on experimental anxiety.

The present study analyzes if estradiol benzoate and/or progesterone interact with desmethylimipramine (DMI) to diminish experimental anxiety. The animal model of anxiety used was the conditioned defensive burying test. Dose response curves for DMI (0.625, 1.25 and 2.5 mg/kg, every 24 h, during 21 days), estradiol benzoate (0.5, 1.0, 2.0 and 4.0 micrograms/rat, 48 h) and progesterone (0.5, 1.0 and 2.0 mg/rat, -4 h) were made in ovariectomized rats. DMI per se decreased dose dependently the cumulative burying time, an effect considered as anxiolytic-like. Progesterone produced a decrease in burying at the highest dose, while estradiol benzoate had no effect on defensive burying. Both, progesterone (0.5 mg/rat) and estradiol benzoate (4.0 micrograms/rat) were able to decrease the cumulative burying behavior when injected with a subthreshold dose of DMI (1.25 mg/kg). In addition, the effect of DMI (1.25 mg/kg) plus the combination of estradiol benzoate and progesterone, sequentially administered (48 h and 4 h before the tests, respectively), also produced a synergistic decrease in burying behavior. In general, the treatments produced no changes in burying behavior latency, neither in spontaneous ambulation or in nociception. It is concluded that DMI synergizes its anxiolytic-like effect when administered with estradiol alone or in combination with progesterone. Present data provide experimental evidence suggesting an interaction between hormones and antidepressants. Results are discussed on the basis of the interaction between steroids and serotonergic or GABAergic receptors.

Orchidectomy sensitizes male rats to the action of diazepam on burying behavior latency: role of testosterone

The anxiolytic-like effect of diazepam (0.0, 0.5, 1.0 mg/kg) on the cumulative burying behavior and the burying behavior latency were compared between intact and castrated male rats. In both groups a clear reduction in cumulative burying, denoting an anxiolytic-like response, was observed. However, castrated males were more sensitive to diazepam in the burying behavior latency, a parameter indicating reactivity. Thus, orchidectomized males showed an increase in burying behavior latency after 1.0 mg/kg diazepam treatment. This dose has no effect on burying behavior latency in intact animals. The higher sensitivity to diazepam on reactivity seems to be androgen dependent because it was reversed by chronic treatment with testosterone propionate (TP, 0.0625 mg/rat for 2 weeks). TP (0.0, 0.0625, 0.125, 0.25, 0.5 and 1.0 mg/rat) produced a dose-dependent reduction in burying behavior after chronic treatment (four injections separated 48 h). At no dose did TP affect burying behavior latency. These results indicate that some actions of diazepam vary in males depending upon the endocrine milieu. Results also support the idea of androgens possessing anxiolytic-like actions.

Estrogens participate in the antidepressant-like effect of desipramine and fluoxetine in male rats

In male rats, the antidepressant-like effect of fluoxetine (FLX) and desipramine (DMI) in the forced swimming test (FST) is reduced by orchidectomy and partially restored by testosterone (T). It is unknown if this modulation of T is produced by its estrogenic metabolites. The objectives of this study were to evaluate if the aromatase inhibitor, formestane, interferes with the antidepressant-like effect of DMI and FLX in intact male rats, and to analyze if 17beta-estradiol (E2) modifies the FST and interacts with the antidepressants in orchidectomized (Orx) males. Intact males received DMI (1.25-5.0 mg/kg) and FLX (2.5-10 mg/kg) alone or in combination with formestane (17.5 mg/kg). Orx rats received E2 (5, 10, 20 and 40 microg/rat) or the combination of E2 [at sub-threshold (5 microg/rat) and optimal (10 microg/rat) doses] plus sub-effective doses of DMI (2.5 mg/kg) or FLX (10 mg/kg). Serum testosterone and estradiol levels were measured in intact-control and -formestane treated animals as well as in castrated males replaced with various doses of E2. Formestane in intact males lacked of an action in the FST, but cancelled the antidepressant-like effect of DMI and FLX. E2 at the supra-physiological doses of 10 and 20 microg/rat produced antidepressant-like effects. E2 at 5 microg/rat (that re-established the levels of this hormone to physiological levels) and at 10 microg/rat restored the antidepressant-like action of DMI and FLX in Orx rats. It was concluded that estrogens participate in the antidepressant-like effect of DMI and FLX in the FST.

Testosterone-dependent antidepressant-like effect of noradrenergic but not of serotonergic drugs.

The main objective of this study was to analyze the effect of testosterone on the actions of antidepressant drugs in the forced swimming test (FST), an animal model that predicts antidepressant effects. In addition, the effect of testosterone propionate (TP) supplementation was evaluated in the same animal model using orchidectomized male rats. Initially, different doses (2.5, 5.0, and 10.0 mg/kg sc, three injections before the test) of desipramine (DMI), fluoxetine (FLX), and clomipramine (CMI) were administered to intact male rats to detect the effective dose in the FST. All drugs (at 10 mg/kg) produced an antidepressant effect, reflected as a reduction of immobility behavior. Neither orchidectomy per se nor TP supplementation (0.5 and 1.0 mg/rat sc, for 10 days) modified the behaviors evaluated in the FST. Orchidectomy blocked the antidepressant effect of DMI, FLX, and CMI (10 mg/kg), while TP supplementation (0.5 mg/rat, for 10 days) restored the antidepressant action of DMI, but not that of CMI or FLX. These findings indicate that testosterone participates in the antidepressant actions of DMI, a noradrenaline reuptake inhibitor, while other gonadal hormones might be involved in the antidepressant effects of the serotonin reuptake inhibitors (SSRIs) like FLX and CMI.

Sex and age differences in the impact of the forced swimming test on the levels of steroid hormones

Compared with the adult disorder, depression in children exhibits differences in its neurobiology, particularly in the HPA axis regulation. The bases of such differences can be evaluated in animal models of depression. The objective of the present study was to determine age and sex differences of Wistar rats in the forced swimming test (FST). The influence of sex and age on corticosterone, estrogens and testosterone serum levels was also determined. Prepubertal rats showed immobility, swimming and climbing behaviors during the pre-test and test sessions. In addition, in the prepubertal animals, no sex differences were found during the pre-test and test sessions. Age comparisons indicated no differences in the female groups, however adult males exhibited more immobility and less swimming than young males, in both FST sessions. The young and female rats showed less immobility behavior and increased levels of estrogens after the FST. The present results indicate that the FST is an animal model suitable to evaluate depressive-like behaviors in prepubertal subjects and to explore behavioral changes related to neurodevelopment.

Aging increases the susceptibility to develop anhedonia in male rats

The objective of this study was to establish the effect of aging on the development of anhedonia, a core feature of depression. Young and old male Wistar rats (of around 3-5 and 12-15 months, respectively) were exposed to a chronic variable stress (CVS) schedule for 3 weeks. CVS produced anhedonia, indicated by a reduction in the intake of a sucrose solution (1%), in 8 out of 23 (35%) young rats and in 19 out of 26 (73%) old rats, implying that old animals are more susceptible to stress and develop anhedonia more readily than young animals. Young and old anhedonic rats showed a similar temporal course in the reduction of sucrose consumption, reaching the anhedonic state after 2 weeks of CVS exposure. Compared with young animals, old rats had lower basal serum testosterone and estradiol levels. The systemic levels of corticosterone did not vary between both age groups. No significant pathological condition was detected in old animals. It is suggested that the higher susceptibility to develop anhedonia in male rats could be associated to neuroendocrine changes consequent to aging.

Aging impairs the antidepressant-like response to citalopram in male rats.

It has been suggested that old depressed patients require longer antidepressant treatments than their young counterparts. The objective of this study was to establish if aging impairs the response to an antidepressant by using an animal model. For this purpose, young and middle-aged male Wistar rats (of around 4 and 14months, respectively) were exposed to a chronic mild stress schedule for 3weeks. After this period, the animals that developed anhedonia, reflected as a reduction in sucrose solution (1%) intake, were treated with citalopram (10mg/kg/day) during 21days while still maintained under the chronic mild stress schedule. Non-stressed animals were included as controls. In young rats citalopram reversed the reduction in sucrose consumption induced by chronic mild stress after one week of treatment, while in middle-aged animals a similar reversion occurred after three weeks. Citalopram did not importantly modify simple water intake in stressed animals or sucrose consumption in non-stressed rats of both ages. The results imply that young rats have a lower latency of onset to the antidepressant-like effect of citalopram than middle-aged animals. The lower sensitivity of middle-aged animals to citalopram could be underlied by their lower levels of gonadal hormones.