Sex Differences in Acute Neuroendocrine Responses to Stressors in Rodents and Humans.

Sex differences in the neuroendocrine response to acute stress occur in both animals and humans. In rodents, stressors such as restraint and novelty induce a greater activation of the hypothalamic-pituitary-adrenal axis (HPA) in females compared to males. The nature of this difference arises from steroid actions during development (organizational effects) and adulthood (activational effects). Androgens decrease HPA stress responsivity to acute stress, while estradiol increases it. Androgenic down-regulation of HPA responsiveness is mediated by the binding of testosterone (T) and dihydrotestosterone (DHT) to the androgen receptor, as well as the binding of the DHT metabolite, 3ß-diol, to the ß form of the estrogen receptor (ERß). Estradiol binding to the α form of the estrogen receptor (ERα) increases HPA responsivity. Studies of human sex differences are relatively few and generally employ a psychosocial paradigm to measure stress-related HPA activation. Men consistently show greater HPA reactivity than women when being evaluated for achievement. Some studies have found greater reactivity in women when being evaluated for social performance. The pattern is inconsistent with rodent studies but may involve the differential nature of the stressors employed. Psychosocial stress is nonphysical and invokes a significant degree of top-down processing that is not easily comparable to the types of stressors employed in rodents. Gender identity may also be a factor based on recent work showing that it influences the neural processing of positive and negative emotional stimuli independent of genetic sex. Comparing different types of stressors and how they interact with gender identity and genetic sex will provide a better understanding of sex steroid influences on stress-related HPA reactivity.

Sex differences in implicit processing of allocentric relationships between objects and location in a Simon task.

Simon tasks reveal implicit processing conflicts that arise when the abstract coding of stimulus position is incongruent with coding for location of the output response. Participants were tested with two versions of a Simon task in a counterbalanced order to examine a potential female bias for attending to object characteristics versus object location. Both tasks used a triangle pointing to the left or right. A simple version presented the triangle in an inner or outer position relative to central fixation. A more complex version included a frame surrounding the inner-outer triangle presentation area in order to introduce additional visual elements for left/right visual processing. When the No Frame version was the first presented, there were no sex differences in the Simon effect in either version, which is consistent with results from other studies that did not provide feedback regarding accuracy. When the initial test was the Frame version, we observed a reverse Simon effect for incongruent triangles presented in the left inner position, with females faster than males to identify the incongruent condition versus the congruent (-59 vs -5 msec). In the No Frame condition that followed, females showed a carryover effect from the previous Frame condition, exhibiting positive Simon effects that were two fold larger than males for identifying incongruent stimuli presented in the left and right outer positions. Similar to previous Simon studies, females showed longer overall reaction times than males (~15%). The difference was not related to the Simon effect and is also found in other types of tasks involving early visual processing of objects with location. Based on sex differences in the Simon effect that emerged following initial experience of the triangle adjoining the frame, the present results support a female bias toward broader integration of objects within the context of location.

Sex related biases for attending to object color versus object position are reflected in reaction time and accuracy.

Processing of visual features related to objects and space relations occurs within separate cortical streams that interact with selective attention. Such separation has implications for cognitive development because the perception of 'what' and 'where' provide a neural foundation for the development of aspects of higher cognition. Thus, a small attentional bias in early development for attending to one aspect over the other might influence subsequent higher cognitive processing in tasks involving object recognition and space relations. We examined 134 men and women for evidence of an inherent sex-related bias for attending to basic perceptual features related to object discrimination versus object position. Each stimulus consisted of a circle located in one of 9 positions within a surrounding frame. Circles were one of three shades of blue or red. These stimuli were used in a match-to-sample paradigm where participants were required to match circles on the basis of color or spatial position. The first stimulus appeared in the center of the screen for 400 msec and the matching stimulus subsequently appeared for 400 msec oriented 5 degrees to the right or left of center. The same stimuli were used to test the perception of color and position, with order of testing counterbalanced across participants. Results showed significantly longer reaction times in females compared with males, with better accuracy to discriminate color when that color was tested before position. Males showed better accuracy when object position was tested before color discrimination. A second experiment employed the same procedure, but enhanced selective attention by adding an endogenous cue that predicted the right or left location for the appearance of the matching stimulus. This manipulation greatly attenuated the sex differences in reaction time and accuracy compared to Experiment 1, suggesting that the sex-related attentional biases are strongly coupled to bottom-up processing. Overall, the sex related attentional biases toward processing object characteristics versus object position location suggest a differential manifestation of biased competition between the weighted systems of dorsal and ventral stream processing. Results are discussed with how a developmental bias in the processing objects versus space relations may contribute to adult cognitive sex differences in humans and animals.

Changes in vasoactive intestinal peptide and arginine vasopressin expression in the suprachiasmatic nucleus of the rat brain following footshock stress.

The neuropeptides, arginine vasopressin (AVP) and vasoactive intestinal polypeptide (VIP) are synthesized by neurons of the suprachiasmatic nucleus (SCN) of the hypothalamus and are important regulators of SCN function. Previous studies have demonstrated that acute exposure to stressors can disrupt circadian activity rhythms, suggesting the possibility of stress-related alterations in the expression of these neuropeptides within SCN neurons. In this study, we examined the effect of intermittent footshock stress on AVP mRNA and heterogeneous nuclear RNA (hnRNA) and VIP mRNA expression in neurons of the SCN. Young adult male Sprague/Dawley rats were subjected to 15 s of scrambled intermittent footshock (0.50 mA pulses, 1 pulse/s, 300 ms duration) every 5 min for 30 min. Animals were sacrificed 75 or 135 min after the onset of stress and brains examined for AVP mRNA and hnRNA, and VIP mRNA using in situ hybridization. Footshock stress increased AVP hnRNA levels at the 75 min time point whereas AVP mRNA was elevated at both the 75 and 135 min time points. In contrast, footshock stress decreased the number of cells expressing VIP mRNA in the SCN without changing hybridization level per cell. These data indicate that the disruptive effect of stress on activity rhythms correlate with alterations in the expression of regulatory peptides within the SCN.

Prenatal ethanol exposure alters core body temperature and corticosterone rhythms in adult male rats.

Ethanol's effects on the developing brain include alterations in morphology and biochemistry of the hypothalamus. To examine the potential functional consequences of ethanol's interference with hypothalamic differentiation, we studied the long-term effects of prenatal ethanol exposure on basal circadian rhythms of core body temperature (CBT) and heart rate (HR). We also examined the late afternoon surge in corticosterone (CORT). Core body temperature and HR rhythms were studied in separate groups of animals at 4, 8, and 20 months of age. The normal late afternoon rise in plasma CORT was examined in freely moving male rats at 6 months of age via an indwelling right atrial cannula. Results showed that the CBT circadian rhythm exhibited an earlier rise after the nadir of the rhythm in fetal alcohol-exposed (FAE) males at all ages compared to controls. At 8 months of age, the amplitude of the CBT circadian rhythm in FAE males was significantly reduced to the level observed in controls at 20 months. No significant effects of prenatal ethanol exposure were observed on basal HR rhythm at any age. The diurnal rise in CORT secretion was blunted and prolonged in 6-month-old FAE males compared to controls. Both control groups exhibited a robust surge in CORT secretion around the onset of the dark phase of the light cycle, which peaked at 7:30 p.m. Whereas FAE males exhibited a linear rise beginning in mid afternoon, which peaked at 9:30 p.m. These results indicate that exposure to ethanol during the period of hypothalamic development can alter the long-term regulation of circadian rhythms in specific physiological systems.

Steroid hormones, receptors, and perceptual and cognitive sex differences in the visual system.

The actions of gonadal steroid hormones induce morphological sex differences in many tissues in the body, including brain. These occur either during development to organize tissues in a sex-specific pattern and/or in adulthood to activate specific cellular pathways. Cellular and morphological changes in the brain, induced by androgens and estrogens, underlie behavioral sex differences in both reproductive and non-reproductive behaviors, including visual perception. A growing body of evidence indicates that some sex differences related to visual perception arise as the result of the organizational actions of gonadal steroid hormones on cerebral cortical pathways involved in visual processing of objects and movement. This review addresses the influence of gonadal steroids on structural, biochemical and morphological changes in tissues in the brain and body. These effects are extended to consider how gonadal hormone effects may contribute to cognitive sex differences across species that are related to processing within the dorsal and ventral visual streams for motion and objects, respectively. Lastly, this review considers the question of how cognitive sex differences related to processing of movement and objects in humans may be reflective of two types of cognitive style that are only superficially related to gender.

Estrogen receptor ß and oxytocin interact to modulate anxiety-like behavior and neuroendocrine stress reactivity in adult male and female rats.

The hypothalamic-pituitary-adrenal (HPA) axis is activated in response to stressors and is controlled by neurons residing in the paraventricular nucleus of the hypothalamus (PVN). Although gonadal steroid hormones can influence HPA reactivity to stressors, the exact mechanism of action is not fully understood. It is known, however, that estrogen receptor ß (ERß) inhibits HPA reactivity and decreases anxiety-like behavior in rodents. Since ERß is co-expressed with oxytocin (OT) in neurons of the PVN, an ERß-selective agonist was utilized to test the whether ERß decreases stress-induced HPA reactivity and anxiety-like behaviors via an OTergic pathway. Adult gonadectomized male and female rats were administered diarylpropionitrile, or vehicle, peripherally for 5days. When tested for anxiety-like behavior on the elevated plus maze (EPM), diarylpropionitrile-treated males and females significantly increased time on the open arm of the EPM compared to vehicle controls indicating that ERß reduces anxiety-like behaviors. One week after behavioral evaluation, rats were subjected to a 20minute restraint stress. Treatment with diarylpropionitrile reduced CORT and ACTH responses in both males and females. Subsequently, another group of animals was implanted with cannulae directed at the lateral ventricle. One week later, rats underwent the same protocol as above but with the additional treatment of intracerebroventricular infusion with an OT antagonist (des Gly-NH2 d(CH2)5 [Tyr(Me)(2), Thr(4)] OVT) or VEH, 20min prior to behavioral evaluation. OT antagonist treatment blocked the effects of diarylpropionitrile on the display of anxiety-like behaviors and plasma CORT levels. These data indicate that ERß and OT interact to modulate the HPA reactivity and the display of anxiety-like behaviors.

Central 5-alpha reduction of testosterone is required for testosterone's inhibition of the hypothalamo-pituitary-adrenal axis response to restraint stress in adult male rats.

In rodents, the hypothalamo-pituitary-adrenal (HPA) axis is controlled by a precise regulatory mechanism that is influenced by circulating gonadal and adrenal hormones. In males, gonadectomy increases the adrenocorticotropic hormone (ACTH) and corticosterone (CORT) response to stressors, and androgen replacement returns the response to that of the intact male. Testosterone (T) actions in regulating HPA activity may be through aromatization to estradiol, or by 5α-reduction to the more potent androgen, dihydrotestosterone (DHT). To determine if the latter pathway is involved, we assessed the function of the HPA axis response to restraint stress following hormone treatments, or after peripheral or central treatment with the 5α-reductase inhibitor, finasteride. Initially, we examined the timecourse whereby gonadectomy alters the CORT response to restraint stress. Enhanced CORT responses were evident within 48 h following gonadectomy. Correspondingly, treatment of intact male rats with the 5α-reductase inhibitor, finasteride, for 48 h, enhanced the CORT and ACTH response to restraint stress. Peripheral injections of gonadectomized male rats with DHT or T for 48 h reduced the ACTH and CORT response to restraint stress. The effects of T, but not DHT, could be blocked by the third ventricle administration of finasteride prior to stress application. These data indicate that the actions of T in modulating HPA axis activity involve 5α-reductase within the central nervous system. These results further our understanding of how T acts to modulate the neuroendocrine stress responses and indicate that 5α reduction to DHT is a necessary step for T action.

Men and women exhibit a differential bias for processing movement versus objects.

Sex differences in many spatial and verbal tasks appear to reflect an inherent low-level processing bias for movement in males and objects in females. We explored this potential movement/object bias in men and women using a computer task that measured targeting performance and/or color recognition. The targeting task showed a ball moving vertically towards a horizontal line. Before reaching the line, the ball disappeared behind a masking screen, requiring the participant to imagine the movement vector and identify the intersection point. For the color recognition task, the ball briefly changed color before disappearing beneath the mask and participants were required only to identify the color shade. Results showed that targeting accuracy for slow and fast moving balls was significantly better in males compared to females. No sex difference was observed for color shade recognition. We also studied a third, dual attention task comprised of the first two, where the moving ball briefly changed color randomly just before passing beneath the masking screen. When the ball changed color, participants were required only to identify the color shade. If the ball didn't change color, participants estimated the intersection point. Participants in this dual attention condition were first tested with the targeting and color tasks alone and showed results that were similar to the previous groups tested on a single task. However, under the dual attention condition, male accuracy in targeting, as well as color shade recognition, declined significantly compared to their performance when the tasks were tested alone. No significant changes were found in female performance. Finally, reaction times for targeting and color choices in both sexes correlated highly with ball speed, but not accuracy. Overall, these results provide evidence of a sex-related bias in processing objects versus movement, which may reflect sex differences in bottom up versus top-down analytical strategies.

Exposure to dexamethasone during late gestation causes female-specific decreases in core body temperature and prepro-thyrotropin-releasing hormone expression in the paraventricular nucleus of the hypothalamus in rats.

Synthetic glucocorticoids (GC) have been used to promote lung development in preterm infants, thereby decreasing respiratory distress syndrome and mortality, yet, concern has arisen from reports that such treatment predisposes individuals to disease in adulthood. Given the variety of preclinical studies that show metabolic and behavioral abnormalities in adulthood following fetal exposure to synthetic GC, we examined the effect of in utero exposure to the synthetic GC, dexamethasone (DEX), on hypothalamic expression of thyrotropin-releasing hormone (TRH) a central neuropeptide involved in mediating behavior and metabolic balance. Pregnant Sprague-Dawley rats were administered 0.4mg/kg DEX on gestational days 18-21. As adults (postnatal day (PD) 60), the offspring were fitted with temperature sensing transmitters allowing real-time monitoring of core body temperature (CBT) across the 24h light dark period. This revealed a significant decrease in CBT throughout the day in prenatal DEX-treated females on estrus and diestrus, but not in male offspring. The reduction in CBT by prenatal DEX exposure was accompanied by a significant decrease in the expression of Trh transcript in the paraventricular nucleus of the hypothalamus (PVN) of female rats at PD 60 and this effect was also present on PD7. There was also a female-specific reduction in the number of preproTRH-immunoreactive (ir) neurons in the PVN, with ppTRH-ir nerve fibers decreases that were present in both male and female offspring. No changes in thyroid hormone (triiodothyronine, T3; thyroxine, T4) were observed in adult offspring, but during development, both males and females (PD14) had lower T3 and T4 levels. These data indicate abnormal expression of TRH results from fetal DEX exposure during late gestation, possibly explaining the decreased CBT observed in the female offspring.

Organizational influence of the postnatal testosterone surge on the circadian rhythm of core body temperature of adult male rats.

The suprachiasmatic nucleus (SCN) of the hypothalamus coordinates physiological and behavioral circadian rhythms such as activity, body temperature, and hormone secretion. Circadian rhythms coordinated by the SCN often show sex differences arising from both organizational and activational effects of gonadal hormones. In males, little is known about the organizational role of testosterone on the circadian regulation of core body temperature (CBT) in adulthood. To explore this, we castrated or sham-operated male rats on the day of birth, and at 4 months of age, implanted them with transmitters that measured CBT rhythms under a 12:12 light/dark cycle. This study revealed a significantly earlier rise in CBT during the light phase in neonatally castrated males. Subsequently, we found that treating neonatally castrated males with testosterone propionate (TP) in adulthood did not reverse the effect of neonatal castration, thus indicating an organizational role for testosterone. In contrast, a single injection of TP at the time of neonatal surgery, to mimic the postnatal surge of testosterone, coupled with TP treatment in adulthood, normalized the circadian rise in CBT. In a final study we examined CBT circadian rhythms in intact adult male and female rats and detected no differences in the rise of CBT during the light phase, although there was a greater overall elevation in female CBT. Together, results of these studies reveal an early organizational role of testosterone in males on the timing of the circadian rise of CBT, a difference that does not appear to reflect "defeminization".

Sex differences in stress-induced hyperthermia in rats: restraint versus confinement.

Studies using restraint to induce psychological stress consistently report the expected hyperthermic response in core body temperature (CBT), but many also report a hypothermic response that precedes the hyperthermia. To understand the conditions that produce hypothermia, and to study sex differences in stress-induced hyperthermia, we measured CBT in male and female rats at 70 and 180 days of age in response to two types of stressors: immobilization through restraint (Plexiglas restrainer) and confinement in a small area (circular wire mesh cylinders that allowed free airflow). Restraint early in the light period induced hypothermia only in 180-day-old males, with no hyperthermia observed during the 30-minute restraint period. Increases in humidity and temperature of the microenvironment due to the larger body weight at this age may contribute to the hypothermia. Hyperthermia during restraint in 70-day-old males was significantly attenuated and delayed in onset compared to the rise in females. All females exhibited a CBT rise of approximately 1.3 degrees C occurring 10-15 min after the onset of restraint. Restraint early in the dark period induced no significant change in CBT in males of either age during immobilization, while females exhibited a small rise of approximately 0.5 degrees C. Confinement early in the light period induced a significant rise of approximately 1.5 degrees C in all groups, with no preceding hypothermia. However, the male response was significantly delayed compared to females. Overall, these results indicate that CBT changes during restraint likely involve both anxiogenic and physiological components, while the marked hyperthermia during confinement is primarily psychological in both sexes.

Inhibition of prolactin secretion from the male rat anterior pituitary by cryptic sequences of prothyrotropin releasing hormone, ProTRH178-199 and ProTRH186-199.

Previous studies have shown that intronic peptide sequences in the prohormone for thyrotropin-releasing hormone (TRH) have physiological actions on pituitary hormone secretion. The aim of this investigation was to examine the effect of the cryptic peptides, prothyrotropin- releasing hormone(178-199) (ProTRH(178-199)) and ProTRH(186-199), on prolactin (PRL) release from the anterior pituitary. Perifusion studies were performed with anterior pituitaries obtained from individual adult male Sprague Dawley rats at 70 90 d of age. Perifusate was collected in 5-min fractions for 25 min prior to peptide administration and for 60 min afterward. Pituitaries were perifused with a single 5 min pulse of either 2, 10, or 40 nM concentrations (peak pulse) of each peptide or the vehicle. Sixty minutes after peptide administration, a 200 mM pulse of potassium chloride was delivered to check tissue viability. Prolactin was measured in the perifusate by radioimmunoassay. Results showed that both peptides induced a significant long-term suppression of prolactin secretion that was still evident at 60 min after peptide exposure. ProTRH(186-199) was similar to ProTRH(178-199) in suppressing prolactin release at the 2 and 40 nM dose, suggesting that the amino acid sequence necessary for prolactin inhibition is contained within the smaller peptide fragment. These data indicate that a cryptic sequence within the proTRH peptide can have biological activity at the level of the anterior pituitary gland in regulating prolactin secretion.

Cognitive efficiency on a match to sample task decreases at the onset of puberty in children.

Electrocortical evidence indicates that a wave of synaptic proliferation occurs in the frontal lobes around the age of puberty onset. To study its potential influence on cognition, we examined 246 children (10-17 years) and 49 young adults (18-22 years) using a match-to-sample type of task to measure reaction times to assess emotionally related information. Based upon the instruction set, subjects made a yes/no decision about the emotion expressed in a face, a word, or a face/word combination presented tachistoscopically for 100 ms. The faces were images of a single individual with a happy, angry, sad or neutral expression. The words were 'happy,' 'angry,' 'sad,' or 'neutral,' In the combined stimulus condition, subjects were asked to decide if the face and word matched for the same emotion. Results showed that compared to the previous year, reaction times were significantly slower for making a correct decision at 11 and 12 years of age in girls and boys, the approximate ages of puberty onset. The peripubertal rise in reaction time declined slowly over the following 2-3 years and stabilized by 15 years of age. Analyses of the performance of 15-17 year olds revealed significantly longer reaction times in females to process both faces and words compared to males. However, this sex difference in late puberty appeared to be transient since it was not present in 18-22 year olds. Given the match-to-sample nature of the task employed, the puberty related increases in reaction time may reflect a relative inefficiency in frontal circuitry prior to the pruning of excess synaptic contacts.