Reproductive and sexual behavior changes in male rats exposed perinatally to picrotoxin.

Previous studies in rats suggested that picrotoxin, a GABA(A) receptor antagonist, may cause long-term changes in male reproductive physiology and behavior in rats exposed during prenatal and postnatal periods. The present study has further examined this phenomenon. Wistar rat dams were dosed subcutaneously with 0.75 mg/kg picrotoxin in saline, or vehicle alone, during the perinatal period (day 19 of gestation, immediately after parturition, and once a day during the first 5 days of lactation). Birth weight and sexual maturation of pups were unchanged; however, plasma testosterone levels and sexual behavior was altered in male offspring. Although fertile, these males showed altered mating behavior in terms of a decrease in the mean number of mounts during a 30-min observation period with normal females. Some showed homosexual behavior when castrated and pretreated with exogenous estrogen. These findings suggest that perinatal exposure to picrotoxin alters sexual dimorphism in the developing rat brain, manifesting as altered reproductive performance and sexual behavior of males.

Reproductive aspects in female rats exposed prenatally to hydrocortisone.

We investigated the effects of hydrocortisone during the prenatal period and its later repercussion on reproductive aspects of female rats. Pregnant rats were treated (s.c.) with hydrocortisone acetate, at 1.5 mg/day on the 17th, 18th, and 19th days of pregnancy. Although the present study was not intended to identify mechanisms of toxicity, the treatment with hydrocortisone in the last period of pregnancy presented no signs of toxicity. The efficacy of the hydrocortisone in reducing the adrenal wet mass and plasma corticosterone levels immediately after delivery in both the treated mothers and in respective pups at birth may indicate impairment of the hypothalamus-pituitary-adrenal axis. In addition, the treatment with hydrocortisone did not interfere in the development of the female descendants until puberty. However, it affected the estrous cycle and fertility. Probably, the prenatal exposure to corticosteroids had altered at least partially the hypothalamus-pituitary-gonadal axis, resulting in the damages observed in adult life. These results indicate that the use of the hydrocortisone at a dose that apparently does not endanger the neonate led to undesirable effects in the adult reproductive phase, resulting in later deleterious alteration of the reproductive physiology in female rats.

A simple high-performance liquid chromatography assay for on-line determination of catecholamines in adrenal gland by direct injection on an ISRP column.

In the present research, assays were improved for the determination of catecholamines in adrenal gland. High-performance liquid chromatography with electrochemical detection was employed for quantitative analysis. The method involved direct injection of acid extract on to a serum albumin dimethylocadecyl-silane (HSA-C18) and the utilization of phosphate buffer (pH = 3.0): methanol (97:3 v/v), 0.025 g heptanosulfonic acid and 0.0025 g EDTA as mobile phase. The detection was obtained using an electrochemical detector L-ECD-6A-Shimadzu with a potential of the 85 mV. Identification was based on retention time. Quantification was performed by automatic peak-area determination. The detection limit is equal to 0.5 ng ml(-1). The HPLC method with electrochemical detection proposed here permits good separation of catecholamines in samples of adrenal gland from rats. The method has various advantages: fast, high precision and good selectivity and do not require sample treatment. The immobilization stress during 5 min did not provoke alteration in catecholamines contains in rat adrenal gland, due to the short time of the stress exposure. This study shows that the catecholamines (norepinephrine and epinephrine) adrenal increased significantly after the acute immobilization stress during 30 min as compared to control group. This increase probably is due to the emotional component of the immobilization stress. In conclusion, our studies suggest an effective participation of the adrenal glands to maintain the homeostasis of organism to the stressful conditions.

Changes in norepinephrine and epinephrine concentrations in adrenal gland of the rats submitted to acute immobilization stress.

Catecholamines act as neurotransmitters and hormones. Studies conducted to understand the synthesis and metabolism of these monoamines during stress have been the main concern of many authors. This work proposes to investigate the time course of changes in epinephrine and norepinephrine concentration in adrenal gland obtained from rats submitted to acute immobilization stress. The results of the present study indicate that acute immobilization stress during 5 and 15 min did not provoke changes in epinephrine and norepinephrine concentrations in adrenal gland in relation to the control group. Such results are justified due to the short time of the stress, showing that the stress did not provoke physiological alteration. The epinephrine and norepinephrine concentrations in adrenal gland increased significantly after the immobilization session in stressed groups during 30 and 50 min as compared to control group. This increase probably is due to the emotional component of the immobilization stress. In this way, we suggested that the immobilization stress provoke increase in the biosynthesis of catecholamines in the adrenal gland from rats. However, the results shows that a maximum increase is reached at 30 min of immobilization stress and then a decrement of catecholamines levels starts at 50 min of the experimental design. This decline in catecholamines level may be consequence of adaptation to stress situations, an increase of the activity of the uptake systems and/or metabolization of catecholamines. In conclusion, these results suggest an effective participation of the adrenal glands to maintain the homeostasis of organism to the stressful conditions.

Cholinergic responses of seminal vesicles isolated from rats exposed perinatally to hydrocortisone.

This study was performed in order to investigate the cholinomimetic response of seminal vesicles isolated from rats treated with hydrocortisone acetate during perinatal life. At the adult phase, the body weight and the wet weight of the seminal vesicle of these animals were unchanged. However, these male rats exhibited a significant reduction in plasma testosterone concentration. A significant increase in the sensitivity of the seminal vesicle to acetylcholine was also observed. Despite this, there was a significant reduction in the maximum contractile response of the organ to this transmitter. These results indicate that exposure to hydrocortisone during the critical period of brain sexual differentiation has a long-term effect on testosterone production of male rats. In addition, physiological levels of cortisone in perinatal life are also essential to support the contractile response pattern of the seminal vesicle to acetylcholine in adult life, probably crucial to the reproductive process.

Effects of neonatal clomiphene citrate on fertility and sexual behavior in male rats.

In order to investigate the participation of estrogen during the period of brain sexual differentiation, male rats were treated with clomiphene citrate in the neonatal phase. Fertility and sexual behavior were assessed during adult life. Sexual maturation, body weight, and wet weight of the testes were unchanged. Although the adult male rats treated with clomiphene in the neonatal phase presented a significant reduction in the frequency of mounts, 90% of these rats were able to mate with normal females, which became pregnant. However, these females exhibited a significantly increased number of pre- and post-implantation losses. When these adult male rats were castrated and received estrogen, 60% presented female sexual behavior (receptive behavior and acceptance of mount). Thus, treatment of pups with clomiphene immediately after birth has a long-term effect on the reproductive physiology and sexual behavior of male rats.

Endocrine disruptors and hypothalamic sexual differentiation

The so-called "endocrine disruptors" have been described as compounds which interfere with the estrogen action in their receptors and may exert a crucial role in the development of the reproductive tract and in the brain sexual differentiation. Thus, conducts and/or exposure to these drugs in the perinatal period that apparently do not endanger the neonate may cause side effects. During embrionary development, the gonads, through discharge of a small quantity of reproductive hormones, will guarantee the phenotype of male or female at birth, as well as actuate in specific areas sexual differentiation of the central nervous system. Several experimental models have shown an interference of drugs acting as endocrine disruptors in hypothalamic sexual differentiation. Thus, reproductive function is impaired by exposure to estrogen in the perinatal life of rats and the mechanisms involved in this effect are distinct for males and females. Perinatal exposure to drugs which may be considered endocrine disrupters may induce an incomplete masculinization and defeminization of the central nervous system. Alterations in these processes, if present, generally are perceived only at puberty or adult reproductive life. These later alterations may include anomalies in the process of fertility or in sexual behavior.

Neonatal inhalatory anesthetic exposure: reproductive changes in male rats.

We investigated the effects of an inhalatory anesthetic (ethyl ether) during the neonatal period of brain sexual differentiation on the later fertility and sexual behavior of male rats. Animals were exposed to ethyl ether immediately after birth. At adulthood, body weight, testes wet weight, and plasma testosterone levels were not affected; however, neonatal exposure to ether showed alterations on male fertility: a decrease in the number of spermatids and spermatozoa, an increase in the transit time of cauda epididymal spermatozoa and a decrease in daily sperm production. An alteration of sexual behavior was also observed: decreased male sexual behavior and appearance of homosexual behavior when the male rats were castrated and pretreated with exogenous estrogen. Probably, the ether delayed or reduced the testosterone peak of the sexual differentiation period, altering the processes of masculinization and defeminization of the hypothalamus. Our results indicate that perinatal exposure to ethyl ether during the critical period of male brain sexual differentiation, acting as endocrine disruptors, has a long-term effect on the fertility and sexual behavior of male rats, suggesting endocrine disruption through incomplete masculinization and defeminization of the central nervous system.

Acute swimming stress induces changes in noradrenergic mechanisms in order to maintain the response pattern of the rat vas deferens to norepinephrine.

This study investigated mechanisms involved in the maintenance of the functional response pattern of the postjunctional alpha(1)-adrenoceptor in vas deferens isolated from rats submitted to acute swimming stress. The plasma corticosterone levels increased approximately three times after the swimming stress in the nontreated rats as well as after swimming stress in the rats pretreated with desipramine (DMI), yohimbine (YO), or DMI with YO. No alteration was detected in the sensitivity to norepinephrine (NE) in the vasa deferentia from the stressed rats or stressed rats treated with DMI or DMI with YO, in relation to their respective control. However, when the vasa deferentia were previously incubated with DMI, a reduction in sensitivity to NE in organs from stressed rats was observed. Vasa deferentia excised from rats pretreated with YO before the swimming stress showed an increase in postjunctional alpha(1)-response that was abolished by prazosin (PZ). Thus, the neuronal uptake, the prejunctional alpha(2)-adrenoceptors (mediating prejunctional inhibition), the occupancy and functional response of the postjunctional alpha(1)-adrenoceptors, and the emotional stress component were very important for the determination of the noradrenergic response pattern in vas deferens from rats submitted to acute swimming stress.