Social interactions and cortisol treatment increase the production of aggressive electrocommunication signals in male electric fish,Apteronotus leptorhynchus

Brown ghost knife fish, Apteronotus leptorhynchus, continually emit a weakly electric discharge that serves as a communication signal and is sensitive to sex steroids. Males modulate this signal during bouts of aggression by briefly (approximately 15 ms) increasing the discharge frequency in signals termed "chirps." The present study examined the effects of short-term (1-7 days) and long-term (6-35 days) male-male interaction on the continuous electric organ discharge (EOD), chirping behavior, and plasma levels of cortisol and two androgens, 11-ketotestosterone (11KT) and testosterone. Males housed in isolation or in pairs were tested for short-term and long-term changes in their EOD frequency and chirping rate to standardized sinusoidal electrical stimuli. Within 1 week, chirp rate was significantly higher in paired fish than in isolated fish, but EOD frequency was equivalent in these two groups of fish. Plasma cortisol levels were significantly higher in paired fish than in isolated fish, but there was no difference between groups in plasma 11KT levels. Among paired fish, cortisol levels correlated positively with chirp rate. To determine whether elevated cortisol can cause changes in chirping behavior, isolated fish were implanted with cortisol-filled or empty Silastic tubes and tested for short-term and long-term changes in electrocommunication signals and steroid levels. After 2 weeks, fish that received cortisol implants showed higher chirp rates than blank-implanted fish; there were no difference between groups in EOD frequency. Cortisol implants significantly elevated plasma cortisol levels compared to blank implants but had no effect on plasma 11KT levels. These results suggest that male-male interaction increases chirp rate by elevating levels of plasma cortisol, which, in turn, acts to modify neural activity though an 11KT-independent mechanism.     

Effects of progesterone, promegestone and RU 486 on glucocorticoid receptor levels in primary cultures of mouse mammary epithelial cells

Mammary epithelial cells isolated from midpregnant mice and cultured on collagen gels contain glucocorticoid receptors whose levels are modulated by a variety of steroids. In the absence of any added steroid to the cell culture medium, the levels of glucocorticoid receptors in the cells decline during culture, which is counteracted by the addition of a variety of glucocorticoid agonists. The effectiveness of the glucocorticoid in preventing the loss of glucocorticoid receptors is in turn counteracted by the addition of the synthetic progestin promegestone and the synthetic antiglucocorticoid RU 486. Of the two, RU 486 is the most potent in antagonizing the effect of cortisol on the GR levels. Promegestone antagonizes the effect of cortisol, too, although higher concentrations are necessary. Progesterone was without a clear effect either as a glucocorticoid agonist or an antagonist. Progesterone, however, was extensively metabolized by mammary epithelial cells in culture. Based on these observations we conclude that in mammary epithelial cells glucocorticoids positively regulate the metabolism of their own receptors and that antiglucocorticoids, such as RU 486 and progestins, can antagonize that effect.     

Recycling and desensitization of glucocorticoid receptors in v-mos transformed cells depend on the ability of nuclear receptors to modulate gene expression

In v-mos transformed cells, glucocorticoid receptor (GR) proteins that bind hormone agonist are not efficiently retained within nuclei and redistribute to the cytoplasmic compartment. These cytoplasmic desensitized receptors cannot be reutilized and may represent trapped intermediates derived from GR recycling. We have used the glucocorticoid antagonist RU486 to examine whether v-mos effects can be exerted on any ligand-bound GR. In the rat 6m2 cell line that expresses a temperature-sensitive p85gag-mos oncoprotein, RU486 is a complete antagonist and suppresses dexamethasone induction of metallothionein-1 mRNA at equimolar concentrations. Using indirect immunofluorescence, we observe efficient nuclear translocation of GR in response to RU486 treatment in either the presence or absence of v-mos oncoproteins. However, in contrast to the redistribution of agonist-bound nuclear receptors to the cytoplasm of v-mos-transformed cells, RU486-bound GRs are efficiently retained within nuclei. Interestingly, withdrawal of RU486 does not lead to efficient depletion of nuclear GR in either nontransformed or v-mos transformed cells. It is only after the addition of hormone agonist to RU486 withdrawn v-mos-transformed cells that GRs are depleted from nuclei and subsequently redistributed to the cytoplasm. Thus, only nuclear GRs that are agonist-bound and capable of modulating gene activity can be subsequently processed and recycled into the cytoplasm.     

Role of the low-affinity glucocorticoid receptor in the regulation of behavior and energy metabolism in the migratory red knot Calidris canutus islandica

Plasma corticosterone increases in association with migratory flight in the red knot Calidris canutus islandica, suggesting that corticosterone may promote migratory activity and/or energy mobilization in this species. This hypothesis is supported by general effects of glucocorticoids, which include stimulation of locomotion and the mobilization of energy depots. We experimentally examined the role of elevated corticosterone levels in the migratory red knot by comparing foraging behavior, flight frequency, and plasma metabolites between vehicle-injected controls and birds treated with RU486, an antagonist to the genomic low-affinity glucocorticoid receptor (GR). We predicted that RU486 treatment would interfere with energy mobilization. However, we expected no effects on flight activity because recent studies suggest that glucocorticoids affect locomotion through a nongenomic receptor. Finally, because glucocorticoids exert permissive effects on food intake, we postulated that RU486 treatment in the red knot would interfere with feeding. Results were consistent with the latter prediction, suggesting that the GR participates in the promotion of hyperphagia, the intense feeding state that is characteristic of the migratory condition. RU486 treatment did not affect flight frequency, suggesting that corticosterone may support migratory activity through a receptor other than the GR. Energy metabolism (as determined through plasma metabolites) was also unaffected by RU486, possibly because energetic demands experienced by captive birds were low.     

Social novelty enhances brain cell proliferation,cell survival,and chirp production in an electric fish,Apteronotus leptorhynchus

For many animals, enriched environments and social interaction promote adult neurogenesis. However, in some cases, the effect is transient, and long‐term environmental stimuli have little benefit for neurogenesis. In electric fish, Apteronotus leptorhynchus, fish housed in pairs for 7 days show higher density of newborn brain cells (cell addition) than isolated fish, but fish paired for 14 days have rates of cell addition similar to isolated controls. We examined whether introduction of social novelty can sustain elevated levels of cell addition and prevent long‐term habituation to social interaction. We also monitored electrocommunication signals (“chirps”) as a measure of the behavioral response to social novelty. We paired fish for 14 days with one continuous partner (no social novelty), two sequential partners changed after 7 days (low novelty) or seven sequential partners changed every 2 days (high novelty). On Day 11, we injected fish with BrdU, sacrificed fish 3 days later and quantified BrdU labeling in the diencephalic periventricular zone. Fish exposed to no novelty had BrdU labeling similar to isolated fish. Fish with low novelty showed small increases in BrdU labeling and those with high novelty had much greater BrdU labeling. Similarly, chirp rates were greater in fish with low novelty than with no novelty and greatest yet in fish with high novelty. By varying the timing of novelty relative to BrdU injection, we showed that social novelty promoted both proliferation and survival of newborn cells. These results indicated that brain cell proliferation and survival is influenced more by social change than simply the presence of social stimuli. © 2012 Wiley Periodicals, Inc. Develop Neurobiol, 2013     

Cell cycle regulation of glucocorticoid receptor function.

Glucocorticoid receptor (GR) nuclear translocation, transactivation and phosphorylation were examined during the cell cycle in mouse L cell fibroblasts. Glucocorticoid-dependent transactivation of the mouse mammary tumor virus promoter was observed in G0 and S phase synchronized L cells, but not in G2 synchronized cells. G2 effects were selective on the glucocorticoid hormone signal transduction pathway, since glucocorticoid but not heavy metal induction of the endogenous Metallothionein-1 gene was also impaired in G2 synchronized cells. GRs that translocate to the nucleus of G2 synchronized cells in response to dexamethasone treatment were not efficiently retained there and redistributed to the cytoplasmic compartment. In contrast, GRs bound by the glucocorticoid antagonist RU486 were efficiently retained within nuclei of G2 synchronized cells. Inefficient nuclear retention was observed for both dexamethasone- and RU486-bound GRs in L cells that actively progress through G2 following release from an S phase arrest. Finally, site-specific alterations in GR phosphorylation were observed in G2 synchronized cells suggesting that cell cycle regulation of specific protein kinases and phosphatases could influence nuclear retention, recycling and transactivation activity of the GR.     

RU486对早孕猕猴海马GR的影响

目的探讨早孕猕猴给予大剂量RU486 3天后海马糖皮质激素受体(GR)的表达变化.方法 15只早孕猕猴随机分为空白对照组、赋形剂组和RU486组.空白对照组不予任何处理,赋形剂组和RU486组分别鼻饲赋形剂和RU486 3天.应用单克隆抗体链菌素亲生物蛋白过氧化酶(SP)免疫组织化学方法观察海马GR的表达情况,并用电子计算机图象分析技术进行处理.结果 RU486组猕猴妊娠终止,该组的海马GR表达显著下降,与对照组的差异有显著性.空白对照组和赋形剂组猕猴妊娠没有终止,其海马GR表达无差异.结论 RU486可使早孕猕猴海马的GR表达下降,推测这可能是该药终止妊娠的中枢作用机理之一.     

Social interaction and cortisol treatment increase cell addition and radial glia fiber density in the diencephalic periventricular zone of adult electric fish, Apteronotus leptorhynchus

In electric fish, Apteronotus leptorhynchus, both long-term social interaction and cortisol treatment potentiates chirping, an electrocommunication behavior that functions in aggression. Chirping is controlled by the diencephalic prepacemaker nucleus (PPn-C) located just lateral to the ventricle. Cells born in adult proliferative zones such as the periventricular zone (PVZ) can migrate along radial glial fibers to other brain regions, including the PPn-C. We examined whether social interactions or cortisol treatment influenced cell addition and radial glia fiber formation by (1) pairing fish (4 or 7 days) or (2) implanting fish with cortisol (7 or 14 days). Adult fish were injected with bromodeoxyuridine 3 days before sacrifice to mark cells that were recently added. Other fish were sacrificed after 1 or 7 days of treatment to examine vimentin immunoreactivity (IR), a measure of radial glial fiber density. Paired fish had more cell addition than isolated fish at 7 days, coinciding temporally with the onset of socially induced increase in chirping behavior. Paired fish also had higher vimentin IR at 1 and 7 days. For both cell addition and vimentin IR, the effect was regionally specific, increasing in the PVZ adjacent to the PPn-C, but not in surrounding regions. Cortisol increased cell addition at 7 days, correlating with the onset of cortisol-induced changes in chirping, and in a regionally specific manner. Cortisol for 14 days increased cell addition, and cortisol for 7 days increased vimentin IR but in a regionally non-specific manner. The correlation between treatment-induced changes in chirping and regionally specific increases in cell addition, and radial glial fiber formation suggests a causal relationship between such behavioral and brain plasticity in adults, but this hypothesis will require further testing.     

In vivo effects of the steroid analogue RU486 on some aspects of intermediary and thyroid metabolism of brook charr, Salvelinus fontinalis.

The glucocorticoid analogue RU486 was administered by intraperitoneal injection to brook charr (Salvelinus fontinalis) to further explore the role of cortisol on aspects of intermediary and thyroid hormone metabolism of the species. RU486 significantly elevated the hepatosomatic index, hepatic G3PDH activity, and hepatic glycogen content, but was without effect on hepatic protein content, hepatic FPBase activity, or plasma glucose concentration. However, the stressor-related increase in plasma glucose concentration that was evident in brook charr 24 h following handling and injection was suppressed in RU486-treated groups. The distribution volume, turnover rates, and metabolic clearance rates of [3H]cortisol were similar in RU486- and vehicle-treated groups. Plasma T3 and T4 concentrations were similar in RU486- and vehicle-treated groups, but hepatic T3 production and hepatic T3 content were lower in RU486-treated fish; TSH had no effect on hepatic T3 content of vehicle-treated brook charr but significantly increased T3 content in the RU486-treated group. These observations support the concept of a role of cortisol in the control of peripheral monodeiodination of T4 in salmonid fish and suggest that RU486 may be a useful drug for evaluating the role of cortisol in fish in vivo.     

The effect of chronic cortisol elevation on urea metabolism and excretion in the rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

The objective of this study was to investigate the possible involvement of cortisol in controlling urea metabolism and excretion in the ammoniotelic rainbow trout (Oncorhynchus mykiss). Trout fitted with dorsal aortic and internal urinary catheters received either no implant (control), or were implanted with coconut oil (sham), cortisol in coconut oil, RU486, a glucocorticoid receptor blocker, in coconut oil, or cortisol+RU486 in coconut oil, and monitored over 72 h. Rainbow trout treated with cortisol (±RU486) had similarly elevated plasma cortisol concentrations that were six fold greater than in control and sham fish. Elevated circulating cortisol concentrations caused a three-fold rise in plasma and urine urea concentrations, which was blocked by RU486. Similarly, a positive correlation between plasma cortisol and plasma urea concentrations was observed in fish treated with cortisol alone but not in fish treated with cortisol+RU486. Cortisol treatment caused an elevation in branchial (two fold higher) and urinary (three fold higher) excretion rates of urea compared to sham-implanted fish, which was prevented by treatment with RU486. However, as branchial and renal clearance were unaffected, there appears to be no stimulation or inhibition of urea excretion mechanisms in the gill or kidney separate from effects due to changes in plasma urea concentrations. Thus, cortisol and glucocorticoid receptors appear to be involved in the regulation of endogenous urea production but not in the control of urea excretory mechanisms in the ammoniotelic trout.Abbreviations GFR glomerular filtration rate - GS glutamine synthetase - O-UC ornithine urea cycle - PEG polyethylene glycol - UFR urine flow rate Communicated by: G. Heldmaier