Androgenic anabolic steroid exposure during adolescence: Ramifications for brain development and behavior

This article is part of a Special Issue “Puberty and Adolescence”.     

Face and voice attractiveness judgments change during adolescence

Attractiveness judgments are thought to underpin adaptive mate choice decisions. We investigated how these judgments change during adolescence when mate choice is becoming relevant. Adolescents aged 11–15 evaluated faces and voices manipulated along dimensions that affect adults' judgments of attractiveness and that are thought to cue mate value. Facial stimuli consisted of pairs of faces that were more or less average, more or less feminine, or more or less symmetric. The adolescents selected the more average, symmetric, and feminine faces as more attractive more often than chance, but judgments of some facial traits differed significantly with rater age and sex, indicating a role of development in judgments of facial cues. Vocal stimuli consisted of pairs of voices manipulated to raise or lower perceived pitch. The older but not younger girls selected the lower-pitched male voices as more attractive at rates above chance, while the younger but not older boys selected the higher-pitched female voices as more attractive. Controlling for rater age, increased pubertal development was associated with increased selection of lower-pitched boys' voices by girls and decreased selection of feminized male faces by boys. Our results are the first to demonstrate that adolescents show somewhat similar attractiveness judgments to adults in age-matched stimuli and that age, sex, and pubertal development have measurable effects on adolescents' attractiveness judgments. They suggest that attractiveness judgments in humans, at least for some traits, are facultatively calibrated to the individual's life stage, only reaching adult values upon sexual maturity when mate choice decisions become relevant.     

Gonadal hormones provide the biological basis for sex differences in behavioral responses to cocaine

Both clinical and rodent studies show sexually dimorphic patterns in the behavioral response to cocaine in all phases of the addiction process (induction, maintenance, and relapse). Clinical and rodent studies also indicate that hormonal fluctuations during the menstrual/estrous cycle modulate cocaine-induced subjective effects in women and locomotor activity in female rats. Evidence suggests that gonadal hormones underlie these observed differences and could be the biological basis of sex-specific differences in cocaine addiction. To study the effects of gonadal hormones on cocaine-induced activity, two approaches have been used. First, studies have examined the role of endogenous hormones through gonadectomy (GDX) and side-by-side comparisons with intact rats. Second, the individual contributions of testosterone, progesterone, and estrogen have been determined by hormone replacement in GDX rats. In this review, we discuss gonadal hormones as the biological basis for the behavioral responses to cocaine, and the clinical implications of these findings.     

Androgens stimulate sex change in protogynous grouper,Epinephelus coioides: spawning performance in sex-changed males

We examined the efficacy of androgens (1.0 mg/kg body mass), testosterone (T), 11-ketotestosterone (11-KT), 17alpha-methyltestosterone (MT), testosterone propionate (TP) or androgen mixture (T, MT and TP in an equal ratio), for induction of sex change in protogynous orange-spotted grouper, Epinephelus coioides. The spawning performance in sex-changed males was also investigated. MT and androgen mixture at a dose of 1.0 mg/kg BW induced a sex transition and completion of spermatogenesis up to the functional male phase. The androgen mixture was most effective. Significantly, higher plasma T levels were found in MT and androgen mixture groups compared to control and other androgen implantation (T, TP or 11-KT) groups. We found that plasma levels of estradiol-17beta (E2) or 11-KT were not different among treated groups. Sex-changed males could successfully fertilize mature eggs. Fertilization and hatching rates were of 23.5-70.4% and 8.4-44.6%, respectively. The data demonstrated that induction of sex change by exogenous androgens in groups could apply to the aquaculture field for seed production.     

Roles of testosterone and amygdaloid LTP induction in determining sex differences in fear memory magnitude

Women are thought to form fear memory more robust than men do and testosterone is suspected to play a role in determining such a sex difference. Mouse cued fear freezing was used to study the sex-related susceptibility and the role of testosterone in fear memory in humans. A 75-dB tone was found to provoke weak freezing, while 0.15-mA and 0.20-mA footshock caused strong freezing responses. No sex differences were noticed in the tone- or footshock-induced (naïve fear) freezing. Following the conditionings, female mice exhibited greater tone (cued fear)-induced freezing than did male mice. Nonetheless, female mice demonstrated indistinctive cued fear freezing across the estrous phases and ovariectomy did not affect such freezing in female mice. Orchidectomy enhanced the cued fear freezing in male mice. Systemic testosterone administrations and an intra-lateral nucleus of amygdala (LA) testosterone infusion diminished the cued fear freezing in orchidectomized male mice, while pretreatment with flutamide (Flu) eradicated these effects. Long-term potentiation (LTP) magnitude in LA has been known to correlate with the strength of the cued fear conditioning. We found that LA LTP magnitude was indeed greater in female than male mice. Orchidectomy enhanced LTP magnitude in males' LA, while ovariectomy decreased LTP magnitude in females' LA. Testosterone decreased LTP magnitude in orchidectomized males' LA and estradiol enhanced LTP magnitude in ovariectomized females' LA. Finally, male mice had lower LA GluR1 expression than female mice and orchidectomy enhanced the GluR1 expression in male mice. These findings, taken together, suggest that testosterone plays a critical role in rendering the sex differences in the cued fear freezing and LA LTP. Testosterone is negatively associated with LA LTP and the cued fear memory in male mice. However, ovarian hormones and LA LTP are loosely associated with the cued fear memory in female mice.     

2d:4d, sex steroid hormones and human psychological sex differences

Studies on 2d:4d, the ratio between the second and the fourth digit, as a possible indicator of prenatal androgen exposure, have failed to produce consistent results. This paper analyzes the relation between 2d:4d, sex steroids and well-documented sex differences in characteristics such as depression, dominance, and aggressive (ART) and non-aggressive adolescent risk-taking (NART) in a comparatively large sample of adolescent boys (N = 301, mean age: 14.4 years) and girls (N = 298, mean age: 14.3 years). Boys had on average a lower 2d:4d than girls (F = 42.15; p < 0.001). With respect to boys, controlling for age and pubertal development (PD), a small but marginally significant positive association was found between 2d:4d and total testosterone (TT) (r = 0.11; p < 0.05). In girls a significant association was found between 2d:4d and SHBG (r = 0.18; p < 0.01). However, relationships between 2d:4d and hormones depended on the phase of the menstrual cycle, with 2d:4d being negatively associated with FT (B = − 0.013; p < 0.05) once a positive association between 2d:4d and FT for girls in the mid-cycle group (B = 0.019; p < 0.01) is taken into account. With respect to sex differences in characteristics, we found evidence of a relationship between 2d:4d and depression in boys (r = − 0.14; p < 0.05) but not between 2d:4d and dominance, ART or NART. No relationships were found between 2d:4d and any of these variables in girls.     

Individual differences in subjective and objective alertness during sleep deprivation are stable and unrelated

This study examines the individual reproducibility of alterations of subjective, objective, and EEG measures of alertness during 27 h of continuous wakefulness and analyzes their interrelationships. Eight subjects were studied twice under similar constant-routine conditions. Scales and performance tasks were administered at hourly intervals to define temporal changes in subjective and objective alertness. The wake EEG was recorded every 2 h, 2 min with eyes open and 2 min with eyes closed. Plasma glucose and melatonin levels were measured to estimate brain glucose utilization and individual circadian phase, respectively. Decrements of subjective alertness and performance deficits were found to be highly reproducible for a given individual. Remarkably, there was no relationship between the impairments of subjective and objective alertness. With increased duration of wakefulness, EEG activity with eyes closed increased in the delta range and decreased in the alpha range, but the magnitudes of these changes were also unrelated. These findings indicate that sleep deprivation has highly reproducible, but independent, effects on brain mechanisms controlling subjective and objective alertness.     

Visual sex discrimination in goldfish: seasonal, sexual, and androgenic influences

The olfactory signals used by goldfish for sexual and aggressive communication have been studied extensively, but little work has addressed the role of other sensory modalities in social communication in this species. We therefore investigated the role that visual stimuli play in sex discrimination and the ability of androgens, which masculinize courtship behavior, to affect behavioral responses toward female visual stimuli. We found that males selectively orient toward female visual stimuli during the breeding season but not outside it, whereas prostaglandin F2-alpha (PGF2alpha)-injected females do not differentially approach male and female visual stimuli, even during the breeding season. Implanting adult females with testosterone (T) and 11-ketotestosterone (KT), however, induced orientation responses toward female visual stimuli similar to those observed in males. These results indicate that visual sexual stimuli are likely important for reproductive signaling in goldfish, potentially helping males identify ovulating females from a distance in a shoal of fish, and that androgens can influence mechanisms associated with orientation responses toward such stimuli.     

Testicular hormone exposure during adolescence organizes flank-marking behavior and vasopressin receptor binding in the lateral septum

Adolescence is a period during which many social behaviors emerge. One such behavior, flank marking, is a testosterone-modulated scent marking behavior that communicates dominance status between adult male Syrian hamsters. Testosterone modulates flank-marking behavior by altering neural transmission of vasopressin within a forebrain circuit. This study tested whether testicular hormones secreted during adolescence play purely a transient activational role in the display of flank-marking behavior, or whether adolescent steroid hormone secretions also cause long-term organizational changes in vasopressin binding within brain regions underlying flank-marking behavior. We tested this hypothesis by manipulating whether testicular secretions were present during adolescent development and then tested for flank-marking behavior and vasopressin receptor binding within the flank-marking neural circuit in young adulthood. Specifically, males were gonadectomized immediately before or after adolescence, replaced with testosterone 6 weeks following gonadectomy in young adulthood, and behavior tested 1 week later. Adult testosterone treatment activated flank-marking behavior only in males that were exposed to testicular hormones during adolescence. In addition, males exposed to testicular hormones during adolescence exhibited significantly less vasopressin receptor binding within the lateral septum than males deprived of adolescent hormones, suggesting that hormone-dependent remodeling of synapses normally occurs in the lateral septum during adolescence. These data highlight the importance of gonadal steroid hormone exposure during adolescence for the organization of neural circuits and social behavior.     

Mechanisms by which neonatal testosterone exposure mediates sex differences in impulsivity in prepubertal rats

Neonatal testosterone, either acting directly or through its conversion to estradiol, can exert organizational effects on the brain and behavior. The goal of the current study was to examine sex differences and determine the role of neonatal testosterone on prefrontal cortex-dependent impulsive choice behavior in prepubertal rats. Male and female prepubertal rats were tested on the delay-based impulsive choice task. Impulsive choice was defined as choosing an immediate small food reward over a delayed large reward. In a first experiment to examine sex differences, males made significantly more impulsive choices than did females. In a second experiment to examine the organizational effects of testosterone, females treated with neonatal testosterone made significantly more impulsive choices than did control females and their performance was indistinguishable from that of control males. In a third experiment to determine if the effect of testosterone on performance is due to the actions of androgens or estrogens through its conversion to estradiol, males treated neonatally with the aromatase inhibitor formestane, which blocks the conversion of testosterone to estradiol, females treated neonatally with the non-aromatizable androgen dihydrotestosterone, and females treated neonatally with estradiol made significantly more impulsive choices than did control females and their performance was indistinguishable from that of control males. Results indicate that male pubertal rats display increased impulsive choice behavior as compared to females, that this sex difference results from organizing actions of testosterone during the neonatal period, and that this effect can result from both androgenic and estrogenic actions.     

Age and sex differences in kidney microRNA expression during the life span of F344 rats

Background

Growing evidence suggests that epigenetic mechanisms of gene regulation may play a role in susceptibilities to specific toxicities and adverse drug reactions. MiRNAs in particular have been shown to be important regulators in cancer and other diseases and show promise as predictive biomarkers for diagnosis and prognosis. In this study, we characterized the global kidney miRNA expression profile in untreated male and female F344 rats throughout the life span. These findings were correlated with sex-specific susceptibilities to adverse renal events, such as male-biased renal fibrosis and inflammation in old age.

Methods

Kidney miRNA expression was examined in F344 rats at 2, 5, 6, 8, 15, 21, 78, and 104 weeks of age in both sexes using Agilent miRNA microarrays. Differential expression was determined using filtering criteria of ≥1.5 fold change and ANOVA or pairwise t-test (FDR <5%) to determine significant age and sex effects, respectively. Pathway analysis software was used to investigate the possible roles of these target genes in age- and sex-specific differences.

Results

Three hundred eleven miRNAs were found to be expressed in at least one age and sex. Filtering criteria revealed 174 differentially expressed miRNAs in the kidney; 173 and 34 miRNAs exhibiting age and sex effects, respectively. Principal component analysis revealed age effects predominated over sex effects, with 2-week miRNA expression being much different from other ages. No significant sexually dimorphic miRNA expression was observed from 5 to 8 weeks, while the most differential expression (13 miRNAs) was observed at 21 weeks. Potential target genes of these differentially expressed miRNAs were identified.

Conclusions

The expression of 56% of detected renal miRNAs was found to vary significantly with age and/or sex during the life span of F344 rats. Pathway analysis suggested that 2-week-expressed miRNAs may be related to organ and cellular development and proliferation pathways. Male-biased miRNA expression at older ages correlated with male-biased renal fibrosis and mononuclear cell infiltration. These miRNAs showed high representation in renal inflammation and nephritis pathways, and included miR-214, miR-130b, miR-150, miR-223, miR-142-5p, miR-185, and miR-296*. Analysis of kidney miRNA expression throughout the rat life span will improve the use of current and future renal biomarkers and inform our assessments of kidney injury and disease.

Electronic supplementary material

The online version of this article (doi:10.1186/s13293-014-0019-1) contains supplementary material, which is available to authorized users.     

How laboratory-raised Japanese monkeys (Macaca fuscata) perceive rotated photographs of monkeys: Evidence for an inversion effect in face perception

Five laboratory-raised Japanese monkeys (Macaca fuscata) were presented various types of photographs of Japanese and rhesus monkeys (Macaca mulatta) in upright, horizontal, and inverted orientations in a sensory-reinforcement experiment. The ratio of the duration of potential viewing time for the photographs which the subjects controlled to the interval between subject-controlled presentations of them (the D/I score) was used as a measure of preference for the photographs. When inverted photographs were presented, the D/I scores were lower than for upright photographs. The difference in D/I scores between photographs of the two species, which indicated discriminability between them, also diminished when the photographs were inverted. The results obtained suggest an inversion effect in face perception in macaque monkeys.     

Blocks and bodies: sex differences in a novel version of the Mental Rotations Test

A novel version of the Mental Rotations Test (MRT) that alternated the standard block figures with three-dimensional human figures was administered to 99 men and 129 women. Women and men differed predictably in their retrospective reports of childhood play and digit ratios, a putative measure of prenatal androgen action. Compared to the block figure items, human figure items on the modified MRT were associated with an improvement in performance in both sexes. However, consistent with the study hypothesis, the enhancing effect of the human figure condition on performance as measured by conventional scores was smaller in men compared to women and not at all evident in men when performance was measured by ratio scores. A closer inspection of the human figures effects on test scores showed performance in women improved for both male and female figure items. In contrast, relative to scores on block figure items, performance in men improved when stimuli were male figures but did not improve when stimuli were female figures. These results add to the evidence that the magnitude of sex differences in scores on the MRT may vary according to the test content and item properties. The findings suggest that online measures of cognitive processing in response to different classes of test stimuli (e.g., animate vs. inanimate objects, self-relevant vs. neutral stimuli) may prove useful in research aimed at understanding the hormonal and social factors contributing to the sex difference in performance on the MRT.     

Developmental differences between singletons and twins in distributions of dental diameter asymmetries

Craniofacial development and behavioral development differs between human twins and singletons in several ways which are related to symmetry development and detectable in adults. In most of those ways, twin zygosity groups do not differ. Here we use distributions of dental diameters, as a model subsystem of craniofacial development, to show that twins, of both zygosities and both sexes, are substantially more symmetrical than singletons. The observed differences are consistent with previous related observations, none of which can readily be explained by any consequence of twin gestation. They seem instead to represent peculiarities of developmental biology familially associated with twinning.     

Chronic stress effects on memory: sex differences in performance and monoaminergic activity

Increasing evidence suggests that the time course of advantageous versus deleterious effects of stress on physiologic function is also apparent in some brain functions, including learning and memory. This article reviews the effects of chronic stress on behavioral performance and, more importantly, shows that sex of the subject, as well as duration and intensity of stress, is an important determinant of the functional/behavioral, neurochemical, and anatomical consequences of the stress. Following chronic stress (7-28 days of restraint, 6 h/day), male and female rats were tested on a visual memory task (object recognition) and two spatial memory tasks (object placement and radial arm maze). At 21 days, stress impaired males on all tasks while females were either enhanced (spatial memory tasks) or not impaired (nonspatial memory tasks). Additionally, the influence of the hypothalamic-pituitary-adrenocortical axis in mediating the sex-specific responses to stress is considered. Behavioral and neurochemical assessments following chronic stress in ovariectomized females, with and without estradiol, suggest that estrogen exerts both organizational and activational influences on the observed sex differences in response to stress. Furthermore, stress differentially affected central transmitter levels in the frontal cortex, hippocampus, and amygdala depending on sex. The possible role of these sex-specific changes in neurotransmitter levels in mediating behavioral differences in response to stress is discussed. While these results are thus far limited to a few studies and require both further investigation and verification, chronic stress appears to be associated with distinct, sex-differentiated behavioral/cognitive and neurochemical responses. We conclude that sex differences must be taken into account when investigating or describing stress and associated sequalae.     

Magnitude of sex differences in dichotomous ossification sequences of the hand and wrist

Among the dichotomous (present/absent:absent/present) ossinication sequences individually ascertained in 3059 boys and girls with at least one but not more than 27 ossification centers of the hand and wrist, 54 such sequences exhibit statistically-significant sex differences in frequency, 32 of them at the 1% confidence level or better. Analyzed by regions (rows and rays, epiphyses and round bones), ten centers, primarily distals and those of the first digit, account for the majority of the significant sex differences.     

Dominant-subordinate relationships in hamsters: sex differences in reactions to familiar opponents

In the majority of mammalian species, males are dominant over and more aggressive than females. In contrast, some reports suggest that female golden hamsters are more aggressive than males but systematic comparisons using the same methods for both sexes are rare. We observed same-sexed pairs of hamsters over repeated trials to assess whether sex differences existed in the level of agonistic behavior and in the development and maintenance of dominant-subordinate relationships with familiar partners. There were no sex differences in measures of agonistic behavior or fear responses (fleeing) during the initial series of three trials on the first day of testing. Following a four-day interval, males that had lost in session 1 showed fearful responses to a familiar dominant male and were not likely to engage in a fight with him. In contrast, females that lost the initial fights were not fearful and fought vigorously with the familiar winner in subsequent encounters. Although the amount of agonistic behavior engaged in by females did decrease over the course of the three sessions, females that lost did not demonstrate an increase in fear, as measured by the latency to flee. Males that lost fights did show increased fear during later trials and sessions. These results suggest that female hamsters are less affected by losing fights than males are and thus that females are less likely than males to develop highly polarized dominant-subordinate relationships. Further work is needed to understand the mechanisms underlying these sex differences.     

The spinal nucleus of the bulbocavernosus: firsts in androgen-dependent neural sex differences

Cell number in the spinal nucleus of the bulbocavernosus (SNB) of rats was the first neural sex difference shown to differentiate under the control of androgens, acting via classical intracellular androgen receptors. SNB motoneurons reside in the lumbar spinal cord and innervate striated muscles involved in copulation, including the bulbocavernosus (BC) and levator ani (LA). SNB cells are much larger and more numerous in males than in females, and the BC/LA target muscles are reduced or absent in females. The relative simplicity of this neuromuscular system has allowed for considerable progress in pinpointing sites of hormone action, and identifying the cellular bases for androgenic effects. It is now clear that androgens act at virtually every level of the SNB system, in development and throughout adult life. In this review we focus on effects of androgens on developmental cell death of SNB motoneurons and BC/LA muscles; the establishment and maintenance of SNB motoneuron soma size and dendritic length; BC/LA muscle morphology and physiology; and behaviors controlled by the SNB system. We also describe new data on neurotherapeutic effects of androgens on SNB motoneurons after injury in adulthood.     

Cognitive performance in rhesus monkeys varies by sex and prenatal androgen exposure

Men and women differ on performance and strategy on several spatial tasks. Rodents display similar sex differences, and manipulations of early hormone exposure alter the direction of these differences. However, most cognitive testing of nonhuman primates has utilized sample sizes too small to investigate sexually differentiated behaviors. This study presents an investigation of sex differences and the effects of prenatal androgen on spatial memory and strategy use in rhesus monkeys. Monkeys prenatally exposed to vehicle, testosterone, or the androgen receptor blocker flutamide performed a search task in which 5 of 12 goal boxes contained food rewards. Spatial consistency and the presence of local landmarks were varied. Performance when both spatial and marker cues were available did not differ by sex or prenatal treatment. Contrary to predictions, females easily solved the task when local markers were removed, and their performance outscored males. Although eliminating spatial consistency and requiring subjects to use local markers impaired performance by all monkeys, females continued to locate correct goal boxes at higher than chance levels and scored better than males. Blocking prenatal androgen exposure in males improved use of local markers. These findings suggest that the tendency to attend to landmarks and to use them in solving spatial problems is typical of females across many species, including rodents, humans, and rhesus monkeys. In rhesus monkeys and rodents, developmental androgen eliminates this specialization. However, these results are the only known example of better performance of females than males when salient markers are removed.