Sex differences in hormonal responses of vasopressin pathways in the rat brain

Vasopressin (AVP) immunoreactivity in cells and projections of the bed nucleus of the stria terminalis (BST) and medial amygdaloid nucleus (MA) depends on gonadal steroids. In addition, the AVP projections from the BST show denser fiber staining in males than in females. To study whether these differences depend on different hormone levels in adulthood, male and female rats were gonadectomized and similarly treated with testosterone for 4 weeks prior to sacrifice. Immunocytochemistry showed that males had significantly more AVP-immunoreactive (AVP-IR) cells in the BST and significantly denser AVP-IR projections from this nucleus to the lateral septum, lateral habenular nucleus, and periaqueductal central gray than did females. The number of AVP-IR cells in the MA nucleus was not statistically different, but denser AVP-IR fiber networks were found in the MA and ventral hippocampus, which receives its input from the MA. No differences were found in the anteroventral portion of the periventricular nucleus and the dorsomedial nucleus of the hypothalamus that receive their AVP innervation from the suprachiasmatic nucleus. These results indicate that the sex difference in the steroid-sensitive AVP pathways depends on other factors besides circulating hormone levels in adulthood.     

Sex differences in neuropeptide distribution in the rat brain

We have investigated possible sex differences in the regional concentrations of neuropeptides in the rat brain. Immunoreactive neurotensin (NT), neurokinin A (NKA), galanin (GAL), calcitonin gene-related peptide (CGRP), substance P (SP) and neuropeptide Y (NPY) were measured by radioimmunoassay in frontal cortex, occipital cortex, hippocampus, striatum, hypothalamus and pituitary in male and female pre- and postpubertal rats. Sex differences were found for NPY (p < 0.001), NT (p < 0.01) and GAL (p < 0.05), in particular in hippocampus, striatum, hypothalamus and pituitary, but not for CGRP, SP and NKA. Results from analysis of neuropeptides in one sex may not be entirely applicable to the other.     

Sex and restraint stress differences in rat metallothionein and Zn levels

Sex differences in serum and liver metallothionein (MT) levels were studied in adult male and female rats. Whereas it was found that female rats had higher hepatic MT levels than male rats in basal, unstressed conditions, no significant differences were found in serum MT levels. Restraint stress increased both serum and liver MT in both sexes. The increase in serum MT was greater in male than in female rats, whereas no significant differences between sexes were found in liver MT content after restraint stress. It is suggested that MT regulation might be sex-dependent and that MT might play some extrahepatic function during stress.     

Sex differences in microglial colonization of the developing rat brain

Microglia are the resident immune cells within the brain and their production of immune molecules such as cytokines and chemokines is critical for the processes of normal brain development including neurogenesis, axonal migration, synapse formation, and programmed cell death. Notably, sex differences exist in many of these processes throughout brain development; however, it is unknown whether a sex difference concurrently exists in the colonization, number, or morphology of microglia within the developing brain. We demonstrate for the first time that the number and morphology of microglia throughout development is dependent upon the sex and age of the individual, as well as the brain region of interest. Males have overall more microglia early in postnatal development [postnatal day (P) 4], whereas females have more microglia with an activated/amoeboid morphology later in development, as juveniles and adults (P30-60). Finally, gene expression of a large number of cytokines, chemokines and their receptors shifts dramatically over development, and is highly dependent upon sex. Taken together, these data warrant further research into the role that sex-dependent mechanisms may play in microglial colonization, number, and function, and their potential contribution to neural development, function, or potential dysfunction.     

Sex differences,laterality, and hormonal regulation of androgen receptor immunoreactivity in rat hippocampus

Sex differences, laterality, and hormonal regulation of androgen receptor (AR) immunoreactivity in rat hippocampal CA1 pyramidal cells were examined using the PG21 antibody. Adult male rats were either castrated or sham-operated at least 2 weeks prior to sacrifice. Gonadally intact females were sacrificed on the day of proestrus. Animals received an injection of either testosterone propionate (TP) or vehicle 2 h prior to sacrifice. Within CA1, both the intensity of staining and the number of AR+ cells were assessed. AR immunostaining was detected in all the groups with marked variation among them. The overall ranking of staining intensity was: gonadally intact males > females given TP > castrated males given TP > females > castrated males given vehicle. The number of AR+cells within subregions of CA1 showed the same basic pattern: among control-treated animals, gonadally intact males have more than females, but castrated males have the least, and acute TP treatment increases the number in both sexes. The increased level of AR immunoreactivity in CA1 of castrated males following acute TP treatment suggests that testicular androgens in adulthood normally increase AR immunoreactivity there, producing a sex difference favoring males in gonadally intact animals. We also found a higher number of AR+ CA1 cells on the left than on the right, but only in gonadally intact males and in females given TP. These results suggest that a laterality of AR distribution in the rat hippocampus may lead to lateralities in hippocampal structure and function.     

Sex differences in long-term stress-induced colonic, behavioural and hormonal disturbances

Functional bowel disorders are more prevalent in women than in men, but the reason for this is unclear. Stressful experiences can increase the risk for or precipitate intestinal dysfunction. Using a model for long-term stress-induced sensitisation in rats, it was investigated whether male and female rats differ in susceptibility for long-term colonic, behavioural and hormonal disturbances following brief but intense stress. Male and female Wistar rats were fitted with chronic electrodes on proximal colon and given either a 15-minute session of foot shocks or no shocks. Two weeks later, rats were exposed to two different novel stressful challenges in the home cage: an electrified prod (day 14) and an 85 dB noise stressor (day 15). Digitalised colonic myoelectric spike burst activity was quantified automatically. Behaviour during prod and noise exposure was scored blindly from videotape. Resting plasma hormone concentrations at the end of the study were determined by radio-immuno assay. Following prod stress on day 14, both male and female preshocked rats showed a greater increase in colonic spike burst frequency than controls, but similar behaviour, and the dynamics of colonic motility differed between sexes. Following noise stress on day 15, only a small change in burst frequency was seen in all rats, but preshocked rats showed less self-grooming behaviour and there was a tendency for preshocked females to show increased noise-induced immobility. Preshocked rats also had lower levels of plasma free thyroxine. While both male and female rats show long-term stress-induced colonic sensitisation and hormonal changes, females show a different activation pattern of colonic motility, and may be more vulnerable for altered behavioural reactivity, following stress.     

Sex differences in lipid metabolism and metabolic disease risk

The ability of nutrients to regulate specific metabolic pathways is often overshadowed by their role in basic sustenance. Consequently, the mechanisms whereby these nutrients protect against or promote a variety of acquired metabolic syndromes remains poorly understood. Premenopausal women are generally protected from the adverse effects of obesity despite having a greater proportion of body fat than men. Menopause is often associated with a transformation in body fat morphology and a gradual increase in the susceptibility to metabolic complications, eventually reaching the point where women and men are at equal risk. These phenomena are not explained solely by changes in food preference or nutrient intake suggesting an important role for the sex hormones in regulating the metabolic fate of nutrients and protecting against metabolic disease pathophysiology. Here, we discuss how differences in the acquisition, trafficking, and subceullular metabolism of fats and other lipid soluble nutrients in major organ systems can create overt sex-specific phenotypes, modulate metabolic disease risk, and contribute to the rise in obesity in the modern sedentary climate. Identifying the molecular mechanisms underpinning sex differences in fat metabolism requires the unravelling of the interactions among sex chromosome effects, the hormonal milieu, and diet composition. Understanding the mechanisms that give rise to sex differences in metabolism will help to rationalize treatment strategies for the management of sex-specific metabolic disease risk factors.     

Sex differences in androgen-regulated expression of cytochrome P450 aromatase in the rat brain

The basis of functional gender differences in adult responsiveness to testosterone (T) is not yet understood. Conversion of T to estradiol by cytochrome P450 aromatase in the medial preoptic area is required for the full expression of male sexual behavior in rats. High levels of aromatase are found in the medial preoptic nucleus (MPN) and in an interconnected group of sexually dimorphic nuclei which mediate masculine sexual behavior. Within this neural circuit, aromatase is regulated by T, acting through an androgen receptor (AR)-mediated mechanism. This arrangement constitutes a feedforward system because T is both the regulator and the major substrate of aromatase. Preoptic aromatase is thus more active in adult males than in females because of normal sex differences in circulating androgen levels. However, the mechanism of enzyme induction also appears to be sexually dimorphic because equivalent physiological doses of T stimulate aromatase to a greater extent in males than in females. Dose-response studies indicate that the sex difference is apparent over a range of circulating T concentrations and constitute a gender difference in T efficacy, but not potency. Sex differences in aromatase correlate with sex differences in nuclear AR concentrations in most regions of the sexually dimorphic neural circuit, but not in MPN. These results suggest that males may have larger populations of target cells in which aromatase is regulated by androgen, but the lack of a gender difference in AR levels in the MPN suggests that differences in post-receptor mechanisms could also be involved. Measurements of aromatase mRNA in androgen-treated gonadectomized rats demonstrate that sex difference in regulation is exerted pretranslationally. Taken together these results demonstrate a sexually dimorphic mechanism that could potentially limit the action of T in females, and may relate to the enhanced expression of T-stimulated sexual behaviors in males.     

Sex differences in ferric nitrilotriacetate-induced lipid peroxidation and nephrotoxicity in mice

Since male A/J mice are much more susceptible to both acute and subacute nephrotoxicity and the carcinogenic effect of ferric nitrilotriacetate than female mice, sex differences in the lipid peroxidation level after ferric nitrilotriacetate use were examined. The effects of orchiectomy and testosterone were also investigated. Male and female A/J mice were given a single intraperitoneal injection of ferric nitrilotriacetate (3 mg of iron/kg of body weight) and then thiobarbituric acid reactivity was determined in the liver and the kidney. Only male mice showed high thiobarbituric acid reactivity after 30 min, with the kidney showing higher activity than the liver. Castrated male mice showed a reduction in thiobarbituric acid reactivity, whereas testosterone-pretreated castrated male or testosterone-pretreated female mice showed increased thiobarbituric acid reactivity. In addition, daily intraperitoneal injections of ferric nitrilotriacetate resulted in the death of all normal male mice within 6 days, whereas all female and castrated male mice survived 3 months of treatment. Thus, male and female mice showed differences in ferric nitrilotriacetate-induced toxicity as reflected in the degree of lipid peroxidation and mortality.     

Sex differences in the regulation of embryonic brain aromatase

Oestrogen formed from androgen by aromatization plays a critical role in the sexual differentiation of the male brain and behaviour. A question which has still to be answered is what regulates the gender-specific changes in aromatase activity forming oestrogen during sensitive periods of brain growth. Using a primary cell culture technique and sexed embryos, we have shown that in the fetal mouse brain, oestrogen formation in the male is neuronal rather than glial and aromatase activity is regionally localized, being higher in the hypothalamus than in the cortex. The aromatase activity measured from cells in culture has the same enzyme binding affinity (apparent K_m 40 nM) as intact brain samples. Neurones developing in the embryonic male brain (embryonic day (ED) 15) contain higher aromatase activity (V_max, 895 fmol/h/mg protein) than the female (V_max, 604). Although a sex difference exists at early stages of embryonic development (ED 13), the embryonic aromatase system is regulated by steroids later in fetal development. The developing aromatase-containing neuroblasts probably form processes which connect to other aromatase neurones. Immunoreactive staining with an aromatase polyclonal antibody identifies an increase in numbers of aromatase-immunoreactive hypothalamic neuronal cell bodies following testosterone treatment. Testosterone treatment also causes both stimulation of neurite growth and branching as well as functional maturation of aromatase neurones. In particular, there is an increase in aromatase activity per neurone as well as a dramatic increase in the number of neurones expressing the enzyme. Both the functional and morphological changes depend on androgen receptor stimulation for several days in vitro. This conclusion is supported by colocalization studies which reveal a high number of fetal hypothalamic aromatase neurones co-expressing androgen receptor. We conclude that testosterone influences the growth of male hypothalamic neurones containing aromatase at a sensitive period of brain development. Endogenous steroid inhibitors of aromatase, probably formed within the neuroglia, also play a role in the control of oestrogen production. An endogenous 5-reduced metabolite of testosterone, 5-androstanedione, is almost as potent in inhibiting neuronal hypothalamic aromatase activity (K_i = 23 nM) as the synthetic non-steroidal inhibitors such as the imidazole, fadrozole, and the triazoles, arimidex and letrozole. It is clear that the oestrogen-forming capacity of the male hypothalamus has the special characteristics and plasticity of regulation which could affect brain differentiation at specific steroid-sensitive stages in ontogeny.     

Sex hormones and neurotransmitters as mediators for sexual differentiation of the brain

Sexual differentiation of the brain is regarded as a model for environment-dependent brain development mediated by systemic hormones and neurotransmitters. Abnormal concentrations of systemic hormones and/or neurotransmitters, if occurring during a critical period of brain development, can lead to permanent developmental disabilities of fundamental processes of life. Such developmental disabilities appear to be avoidable, at least in part, by improving the external, i.e. psychosocial and natural environment, or by correcting abnormalities in the internal, i.e. metabolic and hormonal environment and, particularly, by correcting abnormal neurotransmitter concentrations (and/or turnover rates) during brain development.     

Sex differences in hormonal responses to social conflict in the monogamous California mouse

Monogamous species are usually considered to be less likely to exhibit sex differences in behavior or brain structure. Most previous studies examining sex differences in stress hormone responses have used relatively sexually dimorphic species such as rats. We examined the stress hormone responses of monogamous California mice (Peromyscus californicus) to resident-intruder tests. We also tested males and females under different photoperiods, because photoperiod has been shown to affect both aggression and stress hormone responses. Females, but not males showed a significant increase in corticosterone levels immediately following a resident-intruder test. Males but not females showed elevated corticosterone levels under short days. Females tested in aggression tests also showed a significant increase in plasma oxytocin levels, but only when housed in long days. This was consistent with our observation that females but not males had more oxytocin positive cells in the paraventricular nucleus (PVN) when housed under long days. Our data show that sex differences in glucocorticoid responses identified in other rodents are present in a monogamous species.     

Sex differences of plasma cholinesterase in the rat.

Plasma cholinesterase activity of adult female HAN-Wistar rats was found to be 5.5-fold higher than that of adult male rats kept under constant specified pathogen-free (SPF) conditions up to their 870th day of life.     

Sex differences in estrogen and androgen receptors in hamster brain.

The present study was conducted to measure the levels of estrogen and androgen receptors (ER and AR, receptively) simultaneously in the anterior pituitary (AP), and various brain regions from adult male and proestrous female hamsters. Medial preoptic area (MPOA), medial basal hypothalamus (MBH), lateral hypothalamus (LH), medial forebrain bundle (MFB), and amygdala (AMG) were identified and removed from 200-microns frozen brain sections by the Palkovits punch-out technique. ER and AR were determined by the in vitro binding assay using [3H]-estradiol and [3H]-methyltrienolone as the binding ligands. In males, high levels of AR were found in the MPOA, MBH, and AP. In females, the MPOA, MBH, LH, and AP contained high levels of ER. The males exhibited significantly higher levels of AR than females in the MPOA, MBH, and LH, whereas the ER levels in these areas were higher in females. In males, ER and AR contents in the AP were higher, but the contents in the AMG were lower as compared to those of females. The calculated ER/AR ratio in MPOA, MBH, and LH were lowest in males. On the contrary, the ratio in these areas were highest in females. These data suggest that sex differences in response to estrogen and androgen may in part be due to sex differences in ER and AR contents in specific brain regions.