Fetal endocrine axes mRNA expression levels are related to sex and intrauterine position.

BACKGROUND: The hypothalamic-pituitary-adrenal (HPA) and -gonadal (HPG) axes are two major pathways that connect the neural and endocrine systems in vertebrates. Factors such as prenatal stress and maternal exposure to exogenous steroids have been shown to affect these pathways during fetal development. Another less studied factor is the transfer of hormones across fetuses in multifetal pregnancies. This form of transfer has been shown to influence the morphology, anatomy, physiology, and behavior of the offspring in litter-bearing mammals, an influence termed the intrauterine position (IUP) effect. In this study, we sought to delineate how the IUP effects HPA and HPG brain receptors, peptides, and enzymes (hereafter components) in utero and how these influences may differ between males and females. METHODS: We utilized the unconventional model of culled free-ranging nutria (Myocastor coypus), with its large natural variation. We collected brain tissues from nutria fetuses and quantified the expression of key HPA and HPG components in three brain regions: prefrontal cortex, hypothalamus, and striatum. RESULTS: We found an interaction between sex and IUP in the mineralocorticoid receptor (MR), gonadotropin-releasing hormone receptor (GNRHR), androgen receptor (AR), and estrogen receptor alpha (ESR1). IUP was significant in both gonadotropin-releasing hormone (GnRH) and its receptor GNRHR, but in different ways. In the hypothalamus, fetuses adjacent to same-sex neighbors had higher expression of GnRH than fetuses neighboring the opposite sex. Conversely, in the cortex, GNRHR exhibited the inverse pattern, and fetuses that were neighboring the opposite sex had higher expression levels than those neighboring the same sex. Regardless of IUP, in most components that showed significant sex differences, female fetuses had higher mRNA expression levels than male fetuses. We also found that HPA and HPG components were highly related in the early stages of gestation, and that there was an interaction between sex and developmental stage. In the early stages of pregnancy, female component expression levels were more correlated than males', but in the last trimester of pregnancy, male components were more related to each other than female's. CONCLUSIONS: This study suggests that there are sexually different mechanisms to regulate the HPA and HPG axes during fetal development. Higher mRNA expression levels of endocrine axes components may be a mechanism to help females cope with prolonged androgen exposure over a long gestational period. Additionally, these findings suggest different coordination requirements of male and female endocrine axes during stages of fetal development.

Attentional networks during the menstrual cycle.

The menstrual cycle is characterized partially by fluctuations of the ovarian hormones estradiol (E2) and progesterone (P4), which are implicated in the regulation of cognition. Research on attention in the different stages of the menstrual cycle is eclectic with discrepancies in attention definitions, and the three attentional networks (alerting, orienting and executive) and their interaction were not explored during the menstrual cycle. In the current study, we used the ANT-I (attentional network test - interactions) to examine naturally cycling women (NC) and women using oral contraceptives (OC). We tested their performance at two time points that fit, in natural cycles, the follicular phase and the luteal phase. We found no differences in performance between the two time points (day 4 / day 18) for the OC group: the response pattern replicated known ANT-I findings. However, the NC group showed differences between the two time points. In the follicular phase, responses replicated known ANT-I results, but in the luteal phase, alertness did not interact with executive and orienting networks, resulting in a larger congruency effect (executive network) when attention was not oriented to the target in alerting and no alerting conditions. Results-driven exploratory regression analysis of E2 and P4 suggested that change in P4 from the follicular phase/day 4 to the luteal phase/day 18 was a mediator for the alerting effect found between groups. In conclusion, the alerting state, found with or without alertness manipulation, suggests that there is a progesterone-mediated activation of the alerting system during the luteal phase.

Sex differences in testosterone reactivity and sensitivity in a non-model gerbil.

Although testosterone (T) is a key regulator in vertebrate development, physiology, and behaviour in both sexes, studies suggest that its regulation may be sex-specific. We measured circulating T levels in Baluchistan gerbils (Gerbillus nanus) in the field and in the lab all year round and found no significant sex differences. However, we observed sex differences in circulating T levels following gonadotropin-releasing hormone (GnRH) challenge and T implants in this non-model species. Whereas only males elevated T following a GnRH challenge, females had higher serum T concentrations following T implant insertion. These differences may be a result of different points of regulation along the hypothalamic-pituitary-gonadal (HPG) axis. Consequently, we examined sex differences in the mRNA expression of the androgen receptor (AR) in multiple brain regions. We identified AR and ß-actin sequences in assembled genomic sequences of members of the Gerbillinae, which were analogous to rat sequences, and designed primers for them. The distribution of the AR in G. nanus brain regions was similar to documented expression profiles in rodents. We found lower AR mRNA levels in females in the striatum. Additionally, G. nanus that experienced housing in mixed-sex pairs had higher adrenal AR expression than G. nanus that were housed alone. Regulation of the gerbil HPG axis may reflect evolutionary sex differences in life-history strategies, with males ready to reproduce when receptive females are available, while the possible reproductive costs associated with female T direct its regulation upstream.

A method to determine integrated steroid levels in wildlife claws.

Glucocorticoids act throughout life to regulate numerous physiological and behavioral processes. Their levels are therefore highly labile, reacting to varying conditions and stressors. Hence, measuring glucocorticoids (and other steroids) in wildlife is challenging, and devising methods that are unaffected by the stress of capture and handling should be explored. Here we use the tip of free-ranging chameleons' claws that were cut to allow individual identification, and report a steroids extraction and quantification method. Claw steroids present an integrated level representing the period of claw growth. We found that we could measure corticosterone in small amounts of chameleon claw matrix using commercial EIA kits. Using this method, we learned that in wild male chameleons, claw corticosterone levels were associated with body size. We suggest that claw-testing can potentially provide an ideal matrix for wildlife biomonitoring.

Anandamide protects from low serum-induced apoptosis via its degradation to ethanolamine.

Anandamide (AEA) is a lipid molecule belonging to the family of endocannabinoids. Various studies report neuroprotective activity of AEA against toxic insults, such as ischemic conditions and excitotoxicity, whereas some show that AEA has pro-apoptotic effects. Here we have shown that AEA confers a protective activity in N18TG2 murine neuroblastoma cells subjected to low serum-induced apoptosis. We have demonstrated that the protection from apoptosis by AEA is not mediated via the CB1 receptor, the CB2 receptor, or the vanilloid receptor 1. Interestingly, breakdown of AEA by fatty acid amide hydrolase is required for the protective effect of AEA. Furthermore, the ethanolamine (EA) generated in this reaction is the metabolite responsible for the protective response. The elevation in the levels of reactive oxygen species during low serum-induced apoptosis is not affected by AEA or EA. On the other hand, AEA and EA reduce caspase 3/7 activity, and AEA attenuates the cleavage of PARP-1. Taken together, our results demonstrate a role for AEA and EA in the protection against low serum-induced apoptosis.