M.I. González

The effect of perinatal exposure to estrogens on the sexually dimorphic response to novelty

In this study we investigated the sexually dimorphic anxiety response to a novel environment in the absence of estrogens neonatally or in adulthood. There was a sexual dimorphism in the plus-maze test after the open-field test, females being more active and less anxious. In the absence of estrogens neonatally but not in the adulthood, the activity levels were similar to those shown by females, while the anxiety level was similar to males. These results suggest the need of a normal estrogen environment during the critical period of development for the normal differentiation of female anxiety responses to a novel environment.

Distribution of indoleamines and [3H]paroxetine binding in rat brain regions following acute or perinatal Δ9-tetrahydrocannabinol treatments

The effects of Δ9-tetrahydrocannabinol (Δ9-tetrahydrocannabinol-THC) administration on the central serotoninergic system were evaluated by biochemical assays of tissue levels of indoleamines; a measure of the serotonin (5-HT) innervation was obtained by using [3H]paroxetine as a maker of 5-HT uptake sites. Two different Δ9-THC treatments were chosen, i.e: acute and chronic perinatal maternal exposure. Following acute treatment (5mg/kg), the 5-HT content increased in dorsal hippocampus (+35%), Substantia nigra (+61%) and neostriatum (+62%) but remained unchanged in cingulate cortex, Raphe nuclei, Locus coeruleus and anterior hypothalamus. Endogenous 5-hydroxyindole-3-acetic acid (5-HIAA) decreased in anterior hypothalamus (−23%) and Raphe nuclei (−21%). Following maternal exposure to Δ9-THC (5 mg/kg per day; from gestational day 13 to postnatal day 7), levels of 5-HT were increased in the neostriatum (+22%) but decreased in anterior hypothalamus (−25%), Raphe nuclei (−29%) and Locus coeruleus (−20%) of the litters. Tissue 5-HIAA was increased in anterior hypothalamus (+23%) and Substantia nigra (+48%). There were no changes in 5-HT uptake site density, determined by [3H]paroxetine binding, except for an increase (+50%) in the cingulate cortex of perinatal-treated rats when compared to acutely-treated animals. The present results show that acute and maternal exposure to Δ9-THC produced different effects on the central 5-HT system of the offspring, with a clear regional especifity, but with no changes in the densities of 5-HT uptake sites.

Effect of Δ9-tetrahydrocannabinol on short-term memory in the rat

Abstract We have reported that marihuana and its principal psycoactive compound, Δ 9 -tetrahydrocannabinol ( Δ 9 -THC) produce alterations in several cerebral areas after acute treatment. Based on the involvement of 5-hydroxytryptamine (5-HT) on memory and learning and the reported effects of Δ 9 -THC on short-term memory, we designed an experiment to evaluate the memory performance and its possible relationship with serotonergic alterations after Δ 9 -THC administration. Male Wistar rats received an acute oral dose of THC (5 mg/kg). Short-Term memory was tested on a radial 8-arm maze with a 5 s delay, after 35 days of trainning. The animals were food deprived and adjusted for growth. 5-HT and its metabolite, 5-HIAA, levels were measured in cerebral cortex, dorsal hippocampus, ventral hippocampus, rostral neoestriatum and amygdala basal nucleus, by HPLC-ED. The experiment indicates an impairment of short-term memory in the radial maze test after Δ 9 -THC administration. The control group performed the test without errors, while the treated group made a significant number of errors (Z = 0.019, Mann-Whitney test). This behavioral effect did not seem to be related to serotonergic alterations, as the 5-HT turnover rate was not different between treated and control animals.

Interactions between 5-hydroxytryptamine (5-HT) and testosterone in the control of sexual and nonsexual behaviour in male and female rats

Two 5-hydroxytryptamine2 (5-HT2) agents, ritanserin and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl (DOI) (both at 0.25 mg/kg IP), were administered to castrated males bearing graded testosterone implants (empty, 2.5-, 5-, and 10-mm length) and to normal and neonatally androgenized ovariectomized females bearing 10-mm testosterone implants. The results indicate that testosterone stimulates male sexual behaviour and appears to have a dose-related anxiolytic effect, but no effect on other nonsexual activities. 5-HT and testosterone had opposite effects on male sexual behavior, with ritanserin (5-HT antagonist) enhancing activity in both sexes and DOI (5-HT agonist) inhibiting behaviour in males, the latter being testosterone-dependent. Independent of testosterone, ritanserin reduced locomotion and exploration and increased anxiety in males, while DOI increased locomotion and exploration in both sexes. Ritanserin had a gender-specific effect on anxiety which was independent of testosterone, since in castrated males it was anxiogenic whether they bore testosterone implants or not, while in females it was anxiolytic whether the female were neonatally androgenized (250 micrograms/pup testosterone proprionate [TP] on day 1) or not. These results show that 5-HT and testosterone have opposite effects on male sexual behaviour and these may be interrelated. In adulthood, their effects on nonsexual activities are not inversely related and are independent of each other in contrast to the relationship seen in the neonatal period.

Role of monoamines in the male differentiation of the brain induced by androgen aromatization

Cerebral androgen aromatization has been described as a mechanism responsible for masculinization of the brain, and monoamines seem to be involved in sexual differentiation of the brain. The aim of this study was to investigate the possible implication of monoamines in the masculinization of the brain induced by cerebral androgen aromatization not only in the classic hypothalamic areas but also in some extrahypothalamic ones. For this purpose, 1-day-old male Wistar rats were injected intraventricularly with 5 mg/kg of a suspension of an aromatase inhibitor, LY43578. Saline was administered to male and female control groups. At adulthood, open-field, heterotypical, and homotypical sexual behavior tests were performed and cerebral amines were determined by HPLC-ED. Behavioral tests revealed feminine-like exploratory activity and defecation rate in the treated group, as well as an 89% lordotic response and decreased number of mounts plus intromissions. Testosterone levels were not affected by the treatment. Striatal and limbic serotonergic metabolism showed a sexual dimorphism, higher in males than females, that disappeared in the treated group. From these results, we suggest a possible role of extrahypothalamic serotonin in the mediation of the estrogen-induced mechanisms of behavioral sexual differentiation.

Effect of maternal adrenal deprivation on the content of catecholamines in fetal brain

Previous studies performed in our laboratory showed the importance of the effects that the absence of maternal adrenal hormones have on fetal brain. In the present study we investigated the effect of adrenal deprivation during gestation on the fetal catecholamines development in several cerebral areas. Fetuses from both control and adrenalectomized mothers from the first day of gestation were removed on the 20th embryonary day. Plasma corticosterone levels were significantly lower in the maternal serum of adrenalectomized rats, while the contents were non significantly higher in the adrenalectomized-mothers group of fetuses. Catecholamine contents in diencephalon, metencephalon, mesencephalon and telencephalon were measured by HPLC-ED. The results obtained showed that when the development of the catecholaminergic systems was previous enough to the fetal adrenal function, and under maternal adrenal deprivation conditions, the lack of corticosterone promotes an increase in the level of the catecholamines, as observed in the diencephalic NA, the earlier in maturational process. In those areas where the maturation starts at the same time than the fetal adrenal hypersecretion, no changes were observed. In the cortex, where both DA and NA develop later, the corticosterone produces an inhibition in the proliferation of the catecholaminergic neurons, showing decreased telencephalic levels of both catecholamines.

Extrahypothalamic serotonergic modification after masculinization induced by neonatal gonadal hormones

Exposure to gonadal steroids during the critical period exerts an organizational effect on the CNS. This hormonal effect could be mediated, at least in part, by neurotransmitters. Traditionally, the main place involved in the aminergic sexual differentiation has been the hypothalamus. The aim of this work was to examine the possible long-term effect of cerebral administration of testosterone or estradiol on sexual behavior and hypothalamic/extrahypothalamic monoaminergic systems in the adult rat. For this purpose, female Wistar rats were intraventricularly injected during the first 24 h of life with testosterone (T) or estradiol benzoate (EB) (200 micrograms/kg) (male and female control groups were vehicle treated) and sexual behavior and monoaminergic mediobasal hypothalamic, striatal, and limbic metabolism in adult rats were studied. Receptive behavior was not affected, whereas a masculinizing effect (% mounts) was observed in the animals treated with both gonadal hormones. Only testosterone-treated females showed a male-like serotonergic ratio in corpus striatum and limbic system. A possible extrahypothalamic serotonergic role could be suggested in the mechanisms of sexual differentiation.

Neonatal catecholaminergic influence on behaviour and sexual hormones

There is evidence of a sexually dimorphic effect of serotonin administration during the critical period of sexual differentiation on gonadal hormone secretion in adulthood. To investigate the possible involvement of catecholamines on these mechanisms, we have injected dopamine or noradrenaline intraventricularly into neonatal male and female rats to examine the influence, during the critical period, of this single treatment on the adulthood. Gonadal sex hormone contents, sexually dimorphic behaviours, and catecholaminergic distribution in hypothalamic and extrahypothalamic areas were studied. Both catecholaminergic treatments in females resulted in a reduced striatal dopaminergic activity and an increase in the hypothalamic noradrenergic ratio, while a reduction in the open field activity occurred in the same groups. These results suggest the possible involvement of striatal dopamine and hypothalamic noradrenaline in the differentiation of exploratory activity in females. A reduction in copulatory behaviour was shown in adults of both sexes after neonatal dopaminergic administration, but gonadal hormone levels were not affected in the same way. This indicates the existence of different facets of sexual differentiation, with striatal dopamine and hypothalamic noradrenaline playing important roles in neurobehavioural differentiation.

Influence of amygdala catecholamines on ovarian and adrenal medullary secretion

The amygdaloid complex participates in the modulation of endocrine functions, and contains measurable amounts of noradrenaline (NA) and dopamine (DA). This study examined the contribution of the amygdaloid catecholaminergic systems to the regulation of the adrenal medulla and the ovary. To accomplish this the neurotoxin 6-hydroxydopamine (6-OHDA) was bilaterally injected into the basolateral nucleus of the amygdala (ABL) in cycling rats. The contents of NA and DA in right and left amygdala decreased significantly in lesioned animals with respect to sham lesioned animals, but hypothalamic levels were not different between groups. Administration of 6-OHDA to rats increased the NA, DA and adrenaline (A) contents of the adrenals compared to vehicle treated rats. In addition, lesioned animals showed a significant increase of NA and DA contents in the ovary, although A levels did not differ between groups. Serum oestradiol (O) concentrations were significantly lower in lesioned animals than in controls. These data suggest that the amygdaloid catecholaminergic systems exert an inhibitory effect on catecholamine content of the adrenals and the ovary, and influence the ovarian oestradiol secretion mechanism.

Sexual differences in the neonatal serotonin effect on hormone secretion in rats.

A number of authors in the literature have reported facilitatory or inhibitory effects of serotonin (5-HT) on gonadotropin secretion, sexual hormones content or sexual behavior, but little information has been reported about the possible role of serotonin administered during the critical period of sexual differentiation. To test this possibility, we have injected a 5-HT intraventricularly to neonate male and female rats in order to examine the influence during the critical period of this single treatment on the adult sexual hormone content and sexual behavior. Neonatal administration of 5-HT in the brain decreases significantly estradiol content of adult females, without affecting testosterone level in males. Neither male nor female sexual behavior was affected by 5-HT injection on day 1 of life. These data evidence a sexual difference of serotonin administration during the critical period on gonadal hormones secretion in adulthood.