Sexual differentiation of copulatory behaviour in the male chick requires gonadal steroids.

1. Embryonic injections of 0.3 mg/egg of tamoxifen (TAM), 0.2 mg/egg CI-628 (both antioestrogens), 0.5 mg/egg (ATD (aromatisation inhibitor), or antibodies to oestradiol (E), all suppressed male copulatory activity (MCA) in young male chicks. 2. Embryonic injections with either flutamide (F, androgen antagonist) or high dose of antibodies to testosterone (T) only slightly suppressed MCA. 3. TAM had no effect on embryonic plasma LH levels, 24 and 48 h after injection. 4. It seems that at the embryonic stage oestradiol is required for the normal differentiation of MCA.

The effects of embryonic treatments with gonadal hormones on sexually dimorphic behavior of chicks.

In order to study the role of sex steroids in the differentiation of chick behavior, two groups of experiments were carried out. The first part of the study documented sexual dimorphisms in three behavioral measures in chicks: open-field activity, flocking response, and masculine sexual behavior activated by testosterone (crowing, waltzing, and mating attempts). In the second part, possible organizing influences on these sexually dimorphic behaviors were examined. Male and female embryos were injected with estradiol benzoate (EB) or testosterone propionate (TP). Treatment of males with EB or TP demasculinized all three behaviors. None of the steroid treatments had any effect on the behavior of the females. Plasma testosterone levels of the chicks were not affected by any of these treatments, either before or after testosterone activation. Comb weight was reduced by treatment of male embryos with EB and increased by TP in female embryos, which suggests different mechanism for the development of somatic and behavioral characteristics. The results suggest that exogenous T or E given embryonically can exert similar effects on both sexual behavior and nonreproductive activity of chicks.

Embryonic sex steroids affect mating behavior and plasma LH in adult chickens.

Chicken embryos of both sexes were treated with either antiestrogen (tamoxifen = T), antiandrogen (flutamide = F), aromatization inhibitor (ATD = A), estradiol (E), or oil (control = C). Before puberty, some males of each group were castrated. At puberty, birds were tested under the following regimes: castrated males injected daily with testosterone propionate (CAS + TP) or estradiol benzoate (CAS + EB), intact males (M-INT), intact females (F-INT), and females injected daily with TP (F-TP). In the M-INT and CAS + TP males, E treatment suppressed masculine mating behavior. The embryonic treatments with T, F, and A demasculinized only the frequency of copulations. None of the antihormone treatments caused any masculinization of the sexual activity in the F-TP birds. Untreated males had higher plasma LH than females. The embryonic treatment with E reduced (feminized) the LH levels in CAS + EB birds. This effect was less pronounced in M-INT birds. The results suggest that in chickens, estradiol plays a role in the masculinization of copulatory behavior potential in the developing male embryo. High embryonic estradiol reduces the potential for displaying male sexual behavior at puberty. Feminization of LH secretion requires a high level of estradiol in both embryonic and adult life.

Effect of embryonic treatment with estradiol benzoate on the aggressive behavior of cockerels.

The purpose of this work was to study the effects of early embryonic treatment with estradiol on the aggressive behavior of cockerels, following castration and administration of testosterone or estradiol in adult life. Embryonal estradiol eliminated adult aggressive behavior. However estradiol activation after castration resulted in typical male cock fighting.

Prenatal caffeine causes long lasting behavioral and neurochemical changes.

The effects of prenatal exposure to caffeine were studied on later physical development, behavior and brain neurochemistry. Daily doses (150, 300 or 450 mg/L) of caffeine were given to rat dams during the last week of pregnancy. Prenatal caffeine exposure resulted in a number of behavioral and neurochemical changes in the offspring which were long lasting and dose related. The low dose (150 mg/L) of prenatal caffeine caused hyperactivity in an open-field. The high dose of caffeine caused learning disabilities in complex visual and auditory discrimination learning paradigms while simple motor learning or a spatial orientation task were not affected. Both male and female offspring showed some behavioral effects of caffeine exposure. The medium and high doses of caffeine resulted in weight gain that was observable as early as 35 days of age and increased progressively with age. This weight gain was associated with increased food intake. The neurochemical studies carried out at 2-3 months of age revealed an increase in choline uptake in hippocampus, mainly in the animals treated with the lower doses of caffeine and higher protein concentration (microgram/mg wet tissue) in the cortex or hippocampus of offspring exposed to the higher doses of caffeine. At 15 months of age, choline uptake in the frontal cortex was significantly reduced in the animals prenatally exposed to the 300 and 450 mg/L dose.

The effects of mild maternal stress during pregnancy on the behavior of rat pups.

Mild maternal stress in the form of chronic daily subcutaneous injections of saline or the vehicle for diazepam to pregnant rats was shown to result in some long term, subtle but reliable, changes in the behavior of the offspring. The same vehicle given for the same period of time in the dam's drinking water, without injection had no effect on the development of later behavior of rat pups. Chronic prenatal injections of saline or vehicle for diazepam, used in many experiments as controls for the evaluation of drug effects were shown to have some long lasting behavioral effects in the offspring of the treated dams. The series of experiments reported here compared the offspring of saline or vehicle injected dams to those of uninjected dams on a variety of developmental measurements, an open field behaviour and on learning performance in a complex brightness discrimination maze.

Prenatal monosodium glutamate causes long-lasting cholinergic and adrenergic changes in various brain regions.

Prenatal monosodium glutamate (MSG) given through the mother's diet was found previously to cause behavioral changes in the offspring, including learning disabilities. In the present study, neurochemical parameters were measured in the brains of prenatally exposed rats at various ages throughout development up to adulthood. At 15 days of age, choline uptake and choline acetyltransferase (ChAT) activity in the frontal cortex were significantly reduced (by 80 and 25%, respectively) in MSG-exposed animals, whereas the same cholinergic parameters in hippocampus were not changed. During later development, choline uptake gradually increased, until in adulthood it became significantly higher in MSG-exposed animals than in the controls. This enhancement was found in both males and females. Our previous study showed that only the male offspring were learning disabled. Choline uptake and ChAT activity were enhanced in the hippocampus of adult male animals. Norepinephrine (NE) uptake was reduced (by 25%) in the frontal cortex of males only. There was no change in NE uptake in the hypothalamus.

Prenatal and early postnatal exposure to zimelidine: behavioral, neurochemical and histological findings in rats.

Zimelidine (5 mg/kg/day s.c.) was administered to pregnant rats from day 10 to day 20 of gestation. The development and later open field and learning capacities of their offspring were compared to those of saline injected and untreated dams. The development and behavior of prenatally zimelidine exposed offspring resembled those of the untreated rather than the saline injected group. Pups that were nursed by zimelidine treated mothers, however, showed behavioral deficits compared to those that were nursed by saline injected dams. Prenatal or early postnatal exposure to the 5HT uptake inhibitor zimelidine did not affect 5HT uptake and release measured at 3 months of age. Histological examination of major organs of prenatally zimelidine exposed animals showed no pathological changes.

Effect of teratogenic exposure on the developing brain: research strategies and possible mechanisms.

Some of the problems associated with human behavioral teratology are reviewed and research strategies necessary for the elucidation of the possible prenatal bases for the later occurrence of Learning Disabilities (LD) are suggested. An animal model for LD was created by exposure of female rats during pregnancy or during lactation to diazepam, monosodium glutamate, caffeine, or maternal stress. The gestation period most sensitive to these prenatal insults was established, and some of the neurochemical and histological correlates of the resulting discrimination learning deficits were investigated.

The gonadotropic-axis involvement in the course of the filial following response in the domestic fowl chick.

The detachment process of the domestic chick from its mother, or any other imprinting object occurs between the sixth and tenth week after hatching. The present study (Experiment I), examines whether the detachment process parallels endocrine events that precede prepuberty. Immediately upon hatching, groups of heavy strain chicks were imprinted to a colored foam rubber ball for 72 hours. The bond between these chicks and the imprinting object was then tested, and plasma LH and testosterone were assayed once a week until the chicks were 10 weeks of age; the sexual development of chicks of the same strain was studied at the same time. At the outset of the detachment period (5-7 weeks) an increase in plasma testosterone and a decrease in plasma LH was found. In addition, the comb and testes showed a definite weight increase while the bursa of Fabricius showed a significant decline in weight. In Experiment II, the beginning of the detachment process was induced by injecting 3 to 4 week old chicks with testosterone-propionate, estradiol-benzoate and 5 alpha-dihydrotestosterone. Our evidence therefore appears to demonstrate that testosterone and its metabolites induce the detachment process by the same mechanism used to stimulate sexual behavior in juvenile chicks.

Differential vulnerability of male and female rats to the timing of various perinatal insults.

The results of five experiments showed that exposure to diazepam, hypoxia and monosodium glutamate during the prenatal or early postnatal period of rapid brain development may result in different behavioral consequences depending on the timing of the exposure rather than the nature of the agent. Moreover, male and female offspring may be affected differently by the same agent at different periods of development. Prenatal insults of various kinds impair the later performance of males but not the females in a complex learning task, while postnatal insults seem to affect detrimentally this same behavior in both males and females. The effects of perinatal insults on maze learning and open field activity do not lend themselves to explanation by "feminization" or "masculinization" of behavior caused by interference with prenatal gonadal hormones.

Some long-lasting neurochemical effects of prenatal or early postnatal exposure to diazepam.

Changes in the uptake of various neurotransmitters were measured in the frontal cortex and hippocampus of male and female rats that were exposed to diazepam through the placenta or through the mother's milk during the prenatal or early postnatal period of rapid brain development. Earlier studies from our laboratory showed that early diazepam exposure has long-lasting behavioral consequences. The present results show that prenatally diazepam-exposed rat pups show significant reduction in choline uptake in the frontal cortex at 10 days of age. At 60 days of age, both pre- and postnatally exposed males, but not females, show significant differences from controls in terms of choline uptake, whereas postnatally exposed females whose behavior was shown previously to be profoundly affected by the diazepam exposure showed significant increase in gamma-aminobutyric acid (GABA) uptake in the hippocampus and reduction of 5-hydroxytryptamine (5HT) uptake in the cortex at 60 days of age.

The effect of exposure to diazepam through the placenta or through the mother's milk. Histological findings in slices of rat brain.

Exposure to diazepam during the prenatal or early postnatal developmental period has been reported to result in later behavioural deficits. In the present study morphological changes in the brains of rats that were exposed to diazepam (DZP) prenatally or through the mother's milk postnatally were investigated. The results showed that prolonged prenatal exposure (16 days) to diazepam (10 mg/kg) resulted in characteristic and extensive pathological changes, i.e. gliosis and perivascular cuffing in the brains of the rats. These changes could be observed under the light microscope a long time after exposure to the drug had been terminated. Limiting the prenatal exposure to a single trimester of 7 days reduced somewhat the number of lesions but did not prevent their occurrence. Rats exposed to diazepam postnatally through the mothers' milk showed very few lesions.

Prenatal monosodium glutamate (MSG) treatment given through the mother's diet causes behavioral deficits in rat offspring.

The present study reports various developmental and behavioral changes in the offspring of rat dams that received monosodium glutamate (MSG) in the drinking water all through the second and third trimesters of pregnancy. Three main effects were observed in the MSG exposed offspring: (1) juvenile obesity; (2) reduced general activity levels; (3) a specific type of learning disability in discrimination learning involving choice between simultaneously present positive and negative stimuli.

An ethological analysis of the influence of perinatally-administered diazepam on murine behaviour.

A number of studies have found that the perinatal exposure of rodents to various tranquilizing agents alters their adult behaviour. Given the known anti-aggressive influence of acute doses of diazepam it was hypothesized that, when administered during pregnancy, this drug would change the adult social behaviour. The social interactions of adult male mice whose mothers were treated with diazepam or appropriate controls during pregnancy were video-taped and subjected to an ethological analysis that involved counting the incidences of 43 distinct postures. Prenatal but not postnatal diazepam treatment was associated with a large increase in the incidence of the sideways offensive posture. No significant differences resulted, however, in the case of other postures; in general exposure to prenatal diazepam produced few changes in adult social behaviour.

The effects of exposure to diazepam during various stages of gestation or during lactation on the development and behavior of rat pups.

In the present study we have investigated the effects of diazepam (DZP) (10 mg/kg) treatment of rat dams during different periods of gestation or during lactation on the development and behavior of their offspring. The results show that DZP exposure during different phases of early development has differing effects on later behavior. Exposure during mid-gestation resulted in early and transient hyperactivity, but no learning or memory deficits at 2 months of age were observed. However, both late prenatal and early postnatal exposure to DZP resulted in significant behavioral changes. Late prenatal treatment caused no hyperactivity but resulted in poor performance on the learning and retention of a choice discrimination task, while early postnatal exposure resulted in consistent and lasting hyperactivity and in substantial discrimination learning and retention deficits at 2 months of age.

Feeding and satiation observed in the runway: the effects of d-amphetamine and d-fenfluramine compared.

A paradigm involving feeding to satiety over the course of repeated trials in the runway was used to examine the effects of d-amphetamine (1.0, 1.5 mg/kg) and d-fenfluramine (2.0, 3.0 mg/kg). 1.0 mg/kg d-amphetamine was found to have no significant effect on running performance or feeding in the runway. 1.5 mg/kg d-amphetamine significantly reduced the total food intake during the test but had little impact during the first three trials. In contrast, d-fenfluramine, even at the lower dose and during the initial trials, significantly reduced running performance and feeding to levels normally associated with satiation in the non-drugged animals. The results are discussed in relation to the contrasting modes of action of amphetamine and fenfluramine on food intake.

The effects of postnatal anoxia on behaviour and on the muscarinic and beta-adrenergic receptors in the hippocampus of the developing rat.

Exposure of rats to 25 min anoxia within 24 h following birth caused behavioural as well as biochemical changes during their development and maturity. Following postnatal anoxia, a significant increase in the concentration of the cholinergic muscarinic receptors in the hippocampus was noted at the early stages of development, between 6 and 20 days of age, but reached normal values at 40 days of age. However, at this age, significant increase in the concentration of beta-adrenergic receptors in the hippocampus was found, which remained significantly high during maturity and adulthood, as compared to controls. Rats submitted postnatally to anoxia exhibited hyperactivity in the open field which was maximal at 20-25 days of age and declined towards normal values at 40 days of age. At maturity, between 60 and 80 days of age, these rats showed poor performance in a complex 6-choice discrimination learning but not in simple differential conditioning. Possible correlations between the behavioural and biochemical findings are discussed.

The effects of chronic diazepam treatment on body weight and food intake in rats.

A series of experiments are reported in all of which the effects of chronic (14-18 days) diazepam treatment on body weight were evaluated under various conditions, and one experiment in which weight gain and daily food intake were monitored simultaneously. The effect of diazepam treatment on body weight was compared to that of vehicle treatment in the following groups: (1) young adult females, (2) pregnant females, (3) shock treated young males (4) nonshocked young males. The effect of diazepam treatment on prenatally diazepam exposed and nonexposed males and females was investigated at seven months of age. Food intake was also measured during chronic diazepam and vehicle treatment. The findings show consistent and reliable differences between diazepam and vehicle treatment. Diazepam treated animals eat less in the long run and gain less weight than vehicle treated controls. Prenatal exposure to diazepam enhances the effect of diazepam treatment in adulthood.