Unexpected Reduction of a Coordinated Diazapyridinophane Ligand Bound to Chromium(III) Ion Leading to Delocalization of the Unpaired Electron across Two Isolated Pyridine Units.

In the tetraazamacrocyclic ligand N,N'-dimethyl-2,11-diaza-[3.3](2,6)pyridinophane (L-N4 Me2 ), the two pyridine units are separated from each other by sp3 -hybridized triatomic bridges. Such electronically isolated pyridine moieties are considerably less prone to reductions than di- or triimines. A detailed structural, magnetic, and spectroscopic investigation of the complexes [Cr(L-N4 Me2 )(OAc)2 ] and [Cr(L-N4 Me2 )(OAc)2 ](PF6 ), in combination with theoretical calculations, reveals that the reduced complex must be described as a chromium(III) ion coordinated to the anionic radical ligand (L-N4 Me2 )⋅- rather than a low-spin chromium(II) ion bound to closed-shell ligands. Thus, it is, to the best of our knowledge, only the second example of a stable and structurally characterized metal complex containing a reduced isolated pyridine unit. The stability is attributed to the delocalization of the unpaired electron across the two pyridine units, mediated by their interaction to the metal ion.

MicroRNA-20a inhibits stress-induced cardiomyocyte apoptosis involving its novel target Egln3/PHD3.

Excessive stress, e.g. due to biomechanical overload or ischemia/reperfusion is a potent inductor of cardiomyocyte apoptosis, which contributes to maladaptive remodeling. Despite substantial progress in the understanding of the molecular pathophysiology, many components of the signaling pathways underlying remodeling in general and apoptosis in particular still remain unknown. Recent evidence suggests that microRNAs (miRs) play an important role in the heart's response to increased cardiac stress. To identify novel modulators of stress-dependent remodeling, we conducted a genome-wide miR-screen of mechanically stretched neonatal rat cardiomyocytes (NRCM). Out of 351 miRs, eight were significantly regulated by biomechanical stress, including microRNA-20a, which is part of the miR17-92 cluster. Interestingly, further expression analyses also revealed upregulation of microRNA-20a in an in vitro hypoxia/"reperfusion" model. Given the potential apoptosis-modulating properties of the miR17-92 cluster, we subjected NRCM to hypoxia and subsequent reoxygenation. AdmiR-20a significantly inhibited hypoxia-mediated apoptosis in a dose-dependent fashion, while targeted knockdown of miR-20a in NRCM induced cardiomyocyte apoptosis. Mechanistically, the antiapoptotic effect of miR-20a appears to be mediated through direct targeting and subsequent downregulation of the proapoptotic factor Egln3. Thus, miR-20a is upregulated in acute biomechanical stress as well as hypoxia and inhibits apoptosis in cardiomyocytes. These properties reveal miR-20a as a cardioprotective micro-RNA and a potential target for novel therapeutic strategies to prevent cardiac remodeling.

Estrogen receptor-alpha in the bed nucleus of the stria terminalis regulates social affiliation in male prairie voles (Microtus ochrogaster).

Estrogen receptor alpha (ERalpha) typically masculinizes male behavior, while low levels of ERalpha in the medial amygdala (MeA) and the bed nucleus of the stria terminalis (BST) are associated with high levels of male prosocial behavior. In the males of the highly social prairie vole (Microtus ochrogaster), increasing ERalpha in the MeA inhibited the expression of spontaneous alloparental behavior and produced a preference for novel females. To test for the effects of increased ERalpha in the BST, a viral vector was used to enhance ERalpha expression in the BST of adult male prairie voles. Following treatment, adult males were tested for alloparental behavior with 1-3-day-old pups, and for heterosexual social preference and affiliation. Treatment did not affect alloparental behavior as 73% of ERalpha-BST males and 62.5% of control males were alloparental. Increasing ERalpha in the BST affected heterosexual affiliation, with ERalpha-BST males spending significantly less total time in side-by-side contact with females relative to time spent with control males. ERalpha-BST males did not show a preference for either the familiar or novel female. These findings differed significantly from those reported in ERalpha-MeA enhanced males, where ERalpha inhibited alloparental behavior and produced a preference for a novel female. The findings from this study suggest two things: first, that increased ERalpha in the BST decreases social affiliation and second, that altering ERalpha in different regions of the social neural circuit differentially impacts the expression of social behavior.

Sex steroids are necessary in the second postnatal week for the expression of male alloparental behavior in prairie voles (Microtus ochragaster).

Sex steroids play a significant role in organizing male social behavior, which is associated with low levels of pro-social behavior and high levels of aggression. However, the role of steroids in organizing behavior in highly social males is unclear. The authors tested the hypothesis that low levels of sex steroids facilitate the expression of pro-social behavior in male prairie voles (Microtus ochragaster), predicting that inhibition of testosterone and estradiol would reduce spontaneous-alloparental behavior. Treatment with the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD) or the androgen receptor blocker flutamide, days 8-14, significantly reduced the expression of alloparental behavior in 21-day old males. While both treatments reduced alloparental behavior and increased pup-directed aggression, there were differential treatment effects. Flutamide altered initial response, increasing latency to enter the pup cage and the likelihood of retreat from initial contact. ATD-treated males that were alloparental showed increases in sniffing and latencies to lick and huddle. Results indicate that endogenous steroids play a role in the development of male pro-social behavior and the effects of estrogens and androgens differ.

Photoperiod alters central distribution of estrogen receptor alpha in brain regions that regulate aggression.

Testosterone or its metabolite, estrogen, regulates aggression in males of many mammalian species. Because plasma testosterone levels are typically positively correlated with both aggression and reproduction, aggression is expected to be higher when males are in reproductive condition. However, in some photoperiodic species such as Siberian hamsters (Phodopus sungorus), males are significantly more aggressive in short day lengths when the testes are regressed and circulating testosterone concentrations are reduced. These results led to the formation of the hypothesis that aggression is modulated independently of circulating steroids in Siberian hamsters. Thus, recent studies have been designed to characterize the role of other neuroendocrine factors in modulating aggression. However, aggression may be mediated by testosterone or estrogen despite basal concentrations of these steroids by increasing sensitivity to steroids in specific brain regions. Consistent with this hypothesis, we found that males housed under short days have increased expression of estrogen receptor alpha in the bed nucleus of the stria terminalis, medial amygdala, and central amygdala. Neural activation in response to an aggressive encounter was also examined across photoperiod.

The organizational effects of oxytocin on the central expression of estrogen receptor alpha and oxytocin in adulthood.

BACKGROUND: Previous studies have demonstrated that neonatal manipulation of oxytocin (OT) has effects on the expression of estrogen receptor alpha (ER alpha) and the central production of oxytocin observed in juveniles (at weaning, 21 days of age). The goal of this study was to determine whether the effects of neonatal manipulation of OT last into adulthood, and if the effects differ from those observed during the early postnatal period. On the first day of life, prairie voles (Microtus ochrogaster) received one of three doses of OT (High, 3 microg; Med, 0.3 microg; Low, 0.03 microg), an OT antagonist, or isotonic saline. Another group was handled, but not injected. Then as adults, brains were collected, sectioned, and stained for ER alpha or OT using immunocytochemistry. RESULTS: In females, treatment with OT increased the expression of ER alpha immunoreactivity in the ventral lateral septum (0.03 microg) and the ventromedial nucleus of the hypothalamus and central amygdala (0.3 microg). In males, OT antagonist increased ER alpha expression in the bed nucleus of the stria terminalis. There was no apparent effect of OT on the number of cells producing OT in the paraventricular nucleus of the hypothalamus. CONCLUSION: The current results suggest that neonatal manipulation of OT has long-term organizational effects on the expression of ER alpha in both males and females. The lack of effect on OT neurons in the paraventricular nucleus suggests that some developmental effects of OT previously observed in weanlings do not persist into adulthood. Developmental effects of OT on ER alpha patterns were sexually dimorphic, dose-dependent, and site-specific.

Oxytocin: behavioral associations and potential as a salivary biomarker.

Oxytocin (OT) is a neuropeptide that is produced primarily in the hypothalamus and is best known for its role in mammalian birth and lactation. Recent evidence also implicates OT in social behaviors, including parental behavior, the formation of social bonds, and the management of stressful experiences. OT is reactive to stressors, and plays a role in the regulation of both the central and autonomic nervous system, including effects on immune and cardiovascular function. Knowledge of patterns of OT release would be of value in many fields of science and medicine. However, measurements of OT concentration in blood are infrequently performed, and previous attempts to measure OT in saliva have been unsuccessful. Using a sensitive enzyme immunoassay (EIA) and concentrated samples we were able to detect reproducible changes in salivary OT as a function of lactation and massage. These results indicate that measurements of biologically relevant changes in salivary OT are possible. These results confirm the biological relevance of changes in salivary OT with stressors and support saliva as a noninvasive source to monitor central neuroendocrine function.

Early experience affects the traits of monogamy in a sexually dimorphic manner.

The goal of this study was to examine the effects of early life experiences on the subsequent expression of traits characteristic of social monogamy in prairie voles (Microtus ochrogaster). During cage changes parents and their offspring were either transferred between cages in a cup (zero manipulation, MAN0) or with a gloved hand (one manipulation, MAN1). Following weaning the offspring were tested for alloparental behavior. In adulthood they were tested for the capacity to form partner preferences, behavior in an elevated plus-maze (EPM), and corticosterone levels. MAN0 males (but not females) showed lower levels of alloparental behavior than MAN1 males. MAN0 females (but not males) were less likely to form pair bonds than MAN1 females. MAN0 animals of both sexes were less exploratory in the EPM than MAN1 counterparts. These experiments support the hypothesis that behaviors used to characterize monogamy are vulnerable in a sex-specific manner to early experience.

Developmental effects of oxytocin on neural activation and neuropeptide release in response to social stimuli.

Previous studies have revealed that the neuropeptide hormone oxytocin (OT) has developmental effects on subsequent social behavior and on mechanisms underlying social behavior such as OT neurons and estrogen receptor alpha. This suggests that OT might also have developmental effects on neural responses to social stimuli. This was tested in socially monogamous prairie voles (Microtus ochrogaster) by manipulating OT on the first day of life and then assessing the response to a heterosexual pairing in adulthood. The response to cohabitation was assessed by quantifying neural activation in regions of the brain associated with sociosexual behavior and anxiety using c-Fos immunoreactivity. Additionally, immunocytochemistry was used to label OT and vasopressin neurons and plasma was assayed for both neuropeptides. Treatment effects were evident in females, but not in males. Blockade of OT receptors with an OT antagonist on the first day of life resulted in neural activation of the central amygdala in response to a pairing with a novel male in adulthood. The central amygdala does not normally express c-Fos after a heterosexual pairing in reproductively naïve prairie voles. Treatment effects also were observed in vasopressin immunoreactivity in the SON with OT-treated females showing a decrease.

Effects of stress on parental care are sexually dimorphic in prairie voles.

The effects of stress on parental care are poorly understood, especially in biparental species where males also display care. Data from previous studies in prairie voles, as well as parallels with pair-bonding behavior, suggest the hypothesis that a stressful experience might facilitate parental care in males but not in females. In the present study, male and female prairie voles were exposed to either a 3-min swim stressor or no stressor; 45 min later each animal was tested in a parental care paradigm. Following the parental care test, blood samples were collected and assayed for corticosterone (CORT). After the stressor males, but not females, showed significant changes in parental behavior including significantly more time in kyphosis (arched-back huddling), and a tendency to spend more time licking and grooming pups. In males, CORT levels measured following the parental care test were inversely related to licking and grooming but positively correlated with retrievals. These findings support earlier studies suggesting that the neuroendocrine substrates of parental behavior, as well as the effects of stressors, are sexually dimorphic in this species.

Mechanisms underlying epigenetic effects of early social experience: the role of neuropeptides and steroids.

In mammals the neonatal period is a time of significant social interaction. This is true even in solitary species as females spend a significant amount of time nursing and caring for their offspring. In social species interactions may also include the father, older siblings and extended family members. This period is a time of significant development, including organization of the central nervous system, and therefore a time when the degree and type of social interaction influences the development and expression of social behavior in adulthood. The purpose of this review is to examine the possible mechanisms for the epigenetic effects of early social experience on the subsequent expression of social behavior. We propose that social interactions during the neonatal period organize the subsequent expression of behavior by altering sensitivity to neuropeptides and steroids. Both neuropeptides (e.g. oxytocin and arginine vasopressin) and steroids (e.g. estrogen) regulate or influence the expression of behaviors such as affiliation, aggression, sociosexual behavior, parental behavior, and responses to stress. Therefore, changes in sensitivity to these hormones via reorganization of receptors or changes in hormone production and secretion are potentially powerful mechanisms through which early social experience can mold subsequent social behaviors.

Estrogen receptor alpha and vasopressin in the paraventricular nucleus of the hypothalamus in Peromyscus.

The purpose of this study was to determine the presence of estrogen receptor alpha (ERalpha) and the relationship between neurons that express ERalpha and produce vasopressin (AVP) in the paraventricular nucleus of the hypothalamus (PVN) in new world mice of the genus Peromyscus. Brains were collected from male and female Peromyscus californicus, Peromyscus leucopus, Peromyscus maniculatus, and Peromyscus polionotus, and double labeled for the expression of ERalpha and AVP immunoreactivity (IR). The number of cells expressing ERalpha-IR and AVP-IR was determined in the medial and posterior region of the PVN. The results indicate that Peromyscus is the first taxonomic group reported to have ERalpha widely distributed in the PVN, occurring in both medial and posterior regions of the PVN. While estrogen can regulate the production of AVP, AVP and ERalpha were rarely colocalized. There was, however, a significant inverse relationship between the number of cells that expressed ERalpha-IR and the number expressing AVP-IR. There were no sex differences in the expression of ERalpha-IR or AVP-IR.

Early exposure to oxytocin affects the age of vaginal opening and first estrus in female rats.

Neonatal exposure to exogenous oxytocin (OT) can have long-term effects on the subsequent expression of adult behavior and physiology. Here, we test the prediction that early postnatal exposure to OT can affect the timing of sexual maturation in females, as indicated by the age of vaginal opening and the onset of first estrus. To test this hypothesis, female Sprague-Dawley rats received one of four treatments beginning on the day of birth and continuing for the next 6 days. Three groups received an intraperitoneal injection of one of the following: OT (1 mug/g), an OT antagonist (OTA, 0.1 mug/g) or isotonic saline (vehicle control). The fourth group was handled but not injected. Females were then examined to determine the day of vaginal opening and first estrus. The potential effects of OT on body weight were also measured, with females being weighed on postnatal days 1-7, 70, 91 and 136. Treatment with OT significantly delayed the age of vaginal opening and the onset of first estrus. There was no effect on weight. Results indicate that early exposure to OT can affect the timing and development of female sexual maturation.

Developmental effects of oxytocin on stress response: single versus repeated exposure.

Both exogenous and endogenous oxytocin (OT) are associated with an attenuated stress response. Increased levels of OT in the early postnatal period have been shown to affect behavior and physiology in rodents, and these effects last into adulthood, suggesting an organizational role for OT during development. We investigated the effects of neonatal exposure to OT on the development of the stress response in male and female prairie voles (Microtus ochrogaster). OT or an OT antagonist (OTA) was administered either on postnatal day 1 (single, D1) or days 1-7 (repeated, D1-7) and then on day 8 the response to social isolation was assessed by quantifying ultrasonic vocalizations (USVs) and measuring plasma corticosterone (CORT). Treatment effects were observed only in females. A single treatment with OTA was associated with a decrease in vocalizations, while repeated treatment produced an increase in vocalizations. A single treatment with either saline or OTA increased basal CORT. The results suggest that endogenous OT may be involved in the development of the stress response in females.