Variation In Glucocorticoid Levels Research Articles

Gene expression in the female tree swallow brain is associated with inter- and intra-population variation in glucocorticoid levels

Studies of the evolutionary causes and consequences of variation in circulating glucocorticoids (GCs) have begun to reveal how they are shaped by selection. Yet the extent to which variation in circulating hormones reflects variation in other important regulators of the hypothalamic-pituitary-adrenal (HPA) axis, and whether these relationships vary among populations inhabiting different environments, remain poorly studied. Here, we compare gene expression in the brain of female tree swallows (Tachycineta bicolor) from populations that breed in environments that differ in their unpredictability. We find evidence of inter-population variation in the expression of glucocorticoid and mineralocorticoid receptors in the hypothalamus, with the highest gene expression in a population from an extreme environment, and lower expression in a population from a more consistent environment as well as in birds breeding at an environmentally variable high-altitude site that are part of a population that inhabits a mixture of high and low altitude habitats. Within some populations, variation in circulating GCs predicted differences in gene expression, particularly in the hypothalamus. However, some patterns were present in all populations, whereas others were not. These results are consistent with the idea that some combination of local adaptation and phenotypic plasticity may modify components of the HPA axis affecting stress resilience. Our results also underscore that a comprehensive understanding of the function and evolution of the stress response cannot be gained from measuring circulating hormones alone, and that future studies that apply a more explicitly evolutionary approach to important regulatory traits are likely to provide significant insights.

Personality matters: exploring the relationship between personality and stress physiology in captive African lions

BackgroundConsidering animals as individuals and not as species is becoming increasingly essential to animal welfare management in captive settings. Recent studies on big cat personalities and coping strategies suggest personality can help big cats cope in their surroundings. Yet a large portion of the published literature focuses on understanding either the personality or stress physiology of big cats. Our research shows how integrating an improved understanding of the personality of big cats with stress physiology may enhance welfare, especially for endangered species like African lions. By using a wild cat personality checklist, this study compared the key personality dimensions of 22 African lions with its faecal glucocorticoids and assessed factors influencing their personality and stress physiology.ResultsWe found two reliable personality dimensions for African lions (dominance and agreeableness) and identified key factors (sex, age and location) that may influence their personality. Further, on testing if these factors influenced the stress physiology through variations in glucocorticoid levels, there was no significant difference. However, there was a strong negative association between agreeableness and glucocorticoid levels. These results suggest that the behavioural traits loading positively and higher for agreeableness are associated with lower glucocorticoid stress levels, which may assist a lion to cope with stressors in its surroundings.ConclusionsOur findings highlight this integrated approach of linking personality and stress physiology of big cats can be beneficial for caretakers. For example, during stressful veterinary procedures or in reintroduction programs, recognizing the personality of lions can help in designing or providing them with resources that will alleviate stress. Thus, there is a need for more interdisciplinary approaches that will contribute towards enhancing the individual and overall welfare of big cats.

Infrared thermography is an effective, noninvasive measure of HPA activation

Infrared thermography (IRT) is increasingly applied as a noninvasive technique for measuring surface body temperature changes related to physiological stress. As a basis for validation of IRT as a tool for diagnostic use, we need to assess its potential to measure hypothalamic-pituitary-adrenal (HPA) axis reactivity. We used experimental manipulations of the HPA axis in house sparrows (Passer domesticus), i.e. adrenal tissue responsiveness to exogenous adrenocorticotropin (ACTH) and the efficacy of negative feedback using the synthetic glucocorticoid dexamethasone (DEX), to test whether IRT is an effective tool for measuring HPA reactivity. Experimental birds showed a pronounced decrease in skin temperatures after ACTH injection and an increase in temperature after DEX injection. However, individual variation in glucocorticoid levels were not related to skin temperatures except after ACTH injection in experimental birds. We show that IRT can be used to measure HPA reactivity but that skin-temperature is not a good index for glucocorticoid secretion at baseline levels. These results suggest that while IRT of skin temperatures is a useful, noninvasive measure of HPA axis reactivity under acute activation, this technique might not be suitable for measuring natural variation of circulating glucocorticoid levels.

Effect of food limitation and reproductive activity on fecal glucocorticoid metabolite levels in banded mongooses

BackgroundGlucocorticoids mediate responses to perceived stressors, thereby restoring homeostasis. However, prolonged glucocorticoid elevation may cause homeostatic overload. Using extensive field investigations of banded mongoose (Mungos mungo) groups in northern Botswana, we assessed the influence of reproduction, predation risk, and food limitation on apparent homeostatic overload (n=13 groups, 1542 samples from 268 animals). We experimentally manipulated reproduction and regulated food supply in captive mongooses, and compared their glucocorticoid responses to those obtained from free-living groups.ResultsAt the population level, variation in glucocorticoid levels in free-living mongooses was explained by food limitation: fecal organic matter, recent rainfall, and access to concentrated anthropogenic food resources. Soil macrofauna density and reproductive events explained less and predation risk very little variation in glucocorticoid levels. Reproduction and its associated challenges alone (under regulated feeding conditions) increased glucocorticoid levels 19-fold in a captive group. Among free-living groups, glucocorticoid elevation was seasonal (occurring in late dry season or early wet season when natural food resources were less available), but the timing of peak glucocorticoid production was moderated by access to anthropogenic resources (groups with fewer anthropogenic food sources had peaks earlier in dry seasons). Peak months represented 12- and 16-fold increases in glucocorticoids relative to nadir months with some animals exhibiting 100-fold increases. Relative to the captive group nadir, some free-living groups exhibited 60-fold increases in peak glucocorticoid levels with some animals exhibiting up to 800-fold increases. Most of these animals exhibited 1- to 10-fold increases relative to the captive animal peak.ConclusionsBanded mongooses exhibit seasonal chronic glucocorticoid elevation, associated primarily with food limitation and secondarily with reproduction. Magnitude and duration of this elevation suggests that this may be maladaptive for some animals, with possible fitness consequences. In late dry season, this population may face a convergence of stressors (food limitation, agonistic encounters at concentrated food resources, evictions, estrus, mate competition, parturition, and predation pressure on pups), which may induce homeostatic overload.

On again, off again: Acute stress response and negative feedback together predict resilience to experimental challenges

Abstract Individuals often vary markedly in their ability to cope with stressors, but the drivers of this variation remain poorly understood. Many studies have tested relationships among individual variation in glucocorticoid levels and the response to challenges—often finding inconsistent patterns; however, few have addressed whether variation in the capacity to terminate the stress response through negative feedback is associated with stress resilience. While conceptual models predict that interactions among different components of hypothalamic–pituitary–adrenal (HPA) axis regulation may be important predictors of the phenotypic and fitness effects of stress, we are aware of no previous experimental tests of this hypothesis. We investigate whether individual variation in HPA axis regulation is related to resilience to experimental challenges in free‐living tree swallows (Tachycineta bicolor). We mimicked salient natural challenges by temporarily reducing flight efficiency or increasing perceived predation risk during incubation, and determined whether HPA axis responsiveness prior to treatments predicted resilience. Females that exhibited both a robust HPA axis activation and strong negative feedback were less likely to abandon nests during incubation. Our results suggest that exhibiting a strong HPA axis activation coupled with effective negative feedback may predict stress resilience. Therefore, the ability to turn on and then off the HPA axis efficiently may be important for fitness. Our results also suggest that the interactions between different components of the HPA axis may provide greater insight into differences in stress coping capacity. A plain language summary is available for this article.

Time as tyrant: The minute, hour and day make a difference for corticosterone concentrations in wild nestlings

The hypothalamic-pituitary-adrenal (HPA) axis has been studied extensively in adults, but the HPA axis in early life is not well characterized, and there is an immense amount of unexplained variation in glucocorticoid levels during early life, especially in wild animals. To characterize population-wide natural variation in early-life HPA axis function, we compared plasma corticosterone levels (at baseline and after 30min acute restraint-stress) from seven-day-old nestlings (n=123) from a free-living, marked population of Savannah sparrows (Passerculus sandwichensis). We found a surprising sensitivity of the HPA axis to timing of sample collection across time scales. Even within the accepted 3-min framework to collect baseline samples, time to collect blood had a significant effect on baseline corticosterone concentrations. Daily rhythms also influenced baseline levels, which increased significantly during the relatively short window of sample collection (1100 and 1600). On a broader timeframe, there was a strong effect of hatch date (over a 2month period) on HPA axis responsiveness, where nestlings hatched later in the breeding season had lower stress-induced corticosterone levels than those hatched earlier. The ecophysiological mechanisms and implications of these patterns warrant future investigation; meanwhile this study highlights the critical need to consider, and potentially restrict, time across scales when collecting blood samples from wild birds to assess stress physiology.

Variation in glucocorticoid levels in relation to direct and third-party interactions in a social cichlid fish

In complex animal societies, direct interactions between group members can influence the behavior and glucocorticoid levels of individuals involved. Recently, it has become apparent that third-party group members can influence dyadic interactions, and vice versa. Thus, glucocorticoid levels may vary depending on interactions of other members of the social group. Using the social cichlid fish Neolamprologus pulcher, we examined the relationship between levels of the glucocorticoid hormone cortisol in subordinate females and 1) direct interactions with dominant group members, as well as 2) dyadic interactions between the dominant male and female, in which the subordinate female was not directly involved. Subordinate females that frequently engaged in non-aggressive interactions with dominant females had lower cortisol levels. There was no relationship between subordinate female cortisol and agonistic interactions between the subordinate female and either dominant. Subordinate females had higher cortisol levels when in groups in which the dominant breeding pair behaved agonistically towards each other and performed fewer courtship behaviors. For subordinate females in this species, variation in cortisol levels is associated with their own affiliative behavior, but also can be explained by the broader social context of interactions between dominant members of the group.

Adrenal sensitivity to stress is maintained despite variation in baseline glucocorticoids in moulting seals.

Stressful disturbances activate the hypothalamic-pituitary-adrenal (HPA) axis and result in the release of glucocorticoid (GC) hormones. This characteristic stress response supports immediate energetic demands and subsequent recovery from disturbance. Increased baseline GC concentrations may indicate chronic stress and can impair HPA axis function during exposure to additional stressors. Levels of GCs, however, vary seasonally and with life-history stage, potentially confounding their interpretation. Our objective was to evaluate HPA axis function across variations in baseline GC levels. Northern elephant seals show substantial baseline variation in GC levels during their annual moulting period. We therefore conducted measurements early, in the middle and at the end of moulting; we simulated an acute stressor by administering adrenocorticotrophic hormone and evaluated the changes in circulating hormones and metabolites over the following 2 h. The stress response was characterized by increases in both cortisol and aldosterone (F 7,105 = 153 and 25.3, respectively; P < 0.001). These hormones increased in parallel and the slopes of their relationship varied by study group, suggesting they are regulated in a co--ordinated manner during acute stress in this species. There was no detectable difference in the total release of cortisol or aldosterone among study groups, indicating that the HPA axis remained sensitive to stimulation by adrenocorticotrophic hormone despite varying baseline levels of GCs. Acute stress influenced carbohydrate and fat metabolism in all study groups, but protein catabolism was affected to a far lesser degree. These findings suggest that elephant seals, and potentially other pinniped species, are resilient to moderate variations in baseline GC levels and remain capable of mounting a response to additional stressors.

Primates living outside protected habitats are more stressed: the case of black howler monkeys in the Yucatán Peninsula.

The non-invasive monitoring of glucocorticoid hormones allows for the assessment of the physiological effects of anthropogenic disturbances on wildlife. Variation in glucocorticoid levels of the same species between protected and unprotect areas seldom has been measured, and the available evidence suggests that this relationship may depend on species-specific habitat requirements and biology. In the present study we focused on black howler monkeys (Alouatta pigra), a canopy-dwelling primate species, as a case study to evaluate the physiological consequences of living in unprotected areas, and relate them with intragroup competition and competition with extragroup individuals. From February 2006 to September 2007 we collected 371 fecal samples from 21 adults belonging to five groups (two from protected and three from unprotected areas) in Campeche, Mexico. We recorded agonistic interactions within groups and encounters with other groups (1,200 h of behavioral observations), and determined fecal glucocorticoid metabolite (FGM) concentrations with radioimmunoassays. We used linear mixed models and Akaike's information criterion to choose the best model explaining variation in FGM concentrations between protected and unprotected areas calculated from five categorical variables: habitat type (protected vs. unprotected), participation in agonistic interactions, intergroup encounters, sex and female reproductive state, and season. The best model included habitat type, the interaction between habitat type and agonism, and the interaction between habitat type and season. FGM concentrations were higher in unprotected habitats, particularly when individuals were involved in agonistic interactions; seasonal variation in FGM concentrations was only detected in protected habitats. High FGM concentrations in black howler monkeys living in unprotected habitats are associated with increased within-group food competition and probably associated with exposure to anthropogenic stressors and overall food scarcity. Because persistent high GC levels can be detrimental to health and fitness, populations living in disturbed unprotected areas may not be viable in the long-term.

Brain white matter integrity and cortisol in older men: the Lothian Birth Cohort 1936

Elevated glucocorticoid (GC) levels are hypothesized to be deleterious to some brain regions, including white matter (WM). Older age is accompanied by increased between-participant variation in GC levels, yet relationships between WM integrity and cortisol levels in older humans are underexplored. Moreover, it is unclear whether GC-WM associations might be general or pathway specific. We analyzed relationships between salivary cortisol (diurnal and reactive) and general measures of brain WM hyperintensity (WMH) volume, fractional anisotropy (gFA), and mean diffusivity (gMD) in 90 males, aged 73 years. Significant associations were predominantly found between cortisol measures and WMHs and gMD but not gFA. Higher cortisol at the start of a mild cognitive stressor was associated with higher WMH and gMD. Higher cortisol at the end was associated with greater WMHs. A constant or increasing cortisol level during cognitive testing was associated with lower gMD. Tract-specific bases of these associations implicated anterior thalamic radiation, uncinate, and arcuate and inferior longitudinal fasciculi. The cognitive sequelae of these relationships, above other covariates, are a priority for future study.

Higher 24-h urinary free cortisone, but not cortisol is associated with impaired cortical bone status at the proximal radius in healthy children

Introduction: Impaired bone status is often observed in disease states with overproduction of glucocorticoids. However, whether already moderate variations in glucocorticoid levels may impact on bone is controversially discussed. Since protein intake affects both bone and glucocorticoid status, we herein examined in healthy children and adolescents the association between endogenous glucocorticoids and cortical bone after controlling for protein intake.

Temporal variation in glucocorticoid levels during the resting phase is associated in opposite way with maternal and paternal melanic coloration

Sex-dependent selection can help maintain sexual dimorphism. When the magnitude of selection exerted on a heritable sex trait differs between the sexes, it may prevent each sex to reach its phenotypic optimum. As a consequence, the benefit of expressing a sex trait to a given value may differ between males and females favouring sex-specific adaptations associated with different values of a sex trait. The level of metabolites regulated by genes that are under sex-dependent selection may therefore covary with the degree of ornamentation differently in the two sexes. We investigated this prediction in the barn owl, a species in which females display on average larger black spots on the plumage than males, a heritable ornament. This melanin-based colour trait is strongly selected in females and weakly counter-selected in males indicating sex-dependent selection. In nestling barn owls, we found that daily variation in baseline corticosterone levels, a key hormone that mediates life history trade-offs, covaries with spot diameter displayed by their biological parents. When their mother displayed larger spots, nestlings had lower corticosterone levels in the morning and higher levels in the evening, whereas the opposite pattern was found with the size of paternal spots. Our study suggests a link between daily regulation of glucocorticoids and sex-dependent selection exerted on sexually dimorphic melanin-based ornaments.

Stress, social behaviour, and secondary sexual traits in a male primate

We examined variation in glucocorticoid levels in the mandrill, a brightly coloured primate species, to identify major social influences on stress hormones, and investigate relationships among glucocorticoid levels, testosterone and secondary sexual ornamentation. We collected a total of 317 fecal samples for 16 adult male mandrills over 13 months, including mating and non-mating periods and periods of both dominance rank stability and instability, and compared fecal glucocorticoid levels with dominance rank, rank stability, presence of receptive females, gastro-intestinal parasite infection, fecal testosterone and facial red coloration. Glucocorticoid levels did not vary systematically with dominance rank, but increased when the dominance hierarchy was unstable, and increased in the presence of receptive females. The relationship between dominance rank and glucocorticoid levels changed direction according to the stability of the dominance hierarchy: glucocorticoid levels were higher in subordinate males under stable conditions, but under conditions of instability higher ranking males had higher glucocorticoid levels. The influence of dominance rank also interacted with the presence of receptive females: glucocorticoids were higher in dominant males than in subordinates, but only during mating periods, suggesting that dominant males are more stressed than subordinates during such periods. These findings support previous studies showing that the relationship between glucocorticoids and dominance rank in male baboons is dependent on the social environment. We also found that males with higher glucocorticoids suffered a higher diversity of gastrointestinal parasite infection, in line with evidence that glucocorticoids suppress the immune system in other species. However, we found no support for the stress-mediated immunocompetence handicap hypothesis for the evolution of condition-dependent ornaments: glucocorticoid and testosterone levels were positively related, rather than the negative relationship predicted by the hypothesis, and we found no relationship between red colour and glucocorticoid levels, suggesting that glucocorticoids do not play a role in translating social conditions or physical health into ornament expression in this species.

Infants as costly social tools in male Barbary macaque networks

In mammals, females are the main caretakers of offspring. In a number of species, however, males also care for young, and may spend considerable time with infants. In various nonhuman primates, male interactions with infants appear to benefit mainly the males, who use them to regulate their relationships, particularly with other male group members. We studied whether interactions with infants were related to glucocorticoid output in male Barbary macaques, Macaca sylvanus, at the enclosure ‘La Foret des Singes’, France. We used a combination of behavioural observations, social network analysis and measures of faecal glucocorticoid metabolites as an indicator of physiological stress, and compared the results for the birth season in spring, during a time of intense interactions with infants, with a period in autumn, when males interacted less frequently with infants. Social network analyses revealed that infant carriers had stronger ties with other males than noncarriers. Overall, male relationships were stronger in spring than in autumn. Infant care was unevenly distributed among males and not related to rank, suggesting that the motivation to invest in infant carrying depends on the position in the social network and the current constellation of relationships. Despite the correlation of infant carrying with male interactions, the time spent with infants alone explained most of the variation in glucocorticoid levels, suggesting that access to the male–male network is paid for by an increase in stress hormone levels. Taken together, these results show that Barbary macaque infants can be viewed as costly social tools that facilitate networking among males.

Seasonal and diurnal dynamics of glucocorticoids and behavior in giant pandas

Abstract Because of the suppressive effect of stress on reproduction and health, it is important to evaluate potential stressors that may compromise captive breeding programs for endangered species. However, behavioral and physiological measures of stress are sometimes difficult to interpret, and their relationship to stress can be obscured by factors unrelated to stress, such as seasonal and diurnal patterns. Here we present findings for daily and seasonal variation in glucocorticoid (GC) secretion and behavior from a 6-year study of 2 giant pandas (Ailuropoda melanoleuca). In the American black bear (Ursus americanus), seasonal patterns of corticoid secretion are putatively linked to metabolic demands of hibernation. Although pandas do not hibernate, we have found a similar pattern of GC dynamics. Using radioimmunoassay of urinary GC metabolites, we found seasonal variation in GC levels in an adult female and an adult male panda. As in black bears, winter and spring GC levels were significantly higher t...

GABA receptor modulation of 5-HT neuronal firing: characterization and effect of moderate in vivo variations in glucocorticoid levels

Evidence from electrophysiological studies suggests that 5-HT neuronal firing in the dorsal raphe nucleus (DRN) may be regulated by both GABA A and GABA B receptors. Here, we addressed the question of whether the activity of individual 5-HT neurons is regulated by both GABA A and GABA B receptors. In addition, we examined the concentration-response relationships of GABA A and GABA B receptor activation and determined if GABA receptor regulation of 5-HT neuronal firing is altered by moderate alterations in circulating corticosterone. The activity of 5-HT neurons in the DRN of the rat was examined using in vitro extracellular electrophysiology. The firing of all individual neurons tested was inhibited by both the GABA A receptor agonist 4,5,6,7-tetrahydroisoxazolo-[5,4-c]-pyridin-3-ol hydrochloride (THIP) (25 μM) and the GABA B receptor agonist baclofen (1 μM). Responses to THIP (5, 10, 25 μM) and baclofen (1, 3, 10 μM) were concentration dependent and attenuated by the GABA A and GABA B receptor antagonists, bicuculline (50 μM) and phaclofen (200 μM), respectively. To examine the effects of corticosterone on the sensitivity of 5-HT neurons to GABA receptor activation, experiments were conducted on adrenalectomized animals with corticosterone maintained for two weeks at either a low or moderate level within the normal diurnal range. These changes in corticosterone levels had no significant effects on the 5-HT neuronal response to either GABA A or GABA B recepter activation. The data indicate that the control of 5-HT neuronal activity by GABA is mediated by both GABA A and GABA B receptors and that this control is insensitive to moderate changes in circulating glucocorticoid levels.

Healing of intestinal anastomoses in adrenalectomized rats given corticosterone.

To determine the effect of physiological variations in glucocorticoid levels on wound healing, adrenalectomized rats, implanted with corticosterone pellets of varying concentrations, and sham-operated rats were subjected to ileal transection followed by end-to-end anastomosis. Adrenalectomized animals with plasma corticosterone levels less than 2.5 micrograms/dl suffered 25% mortality. In rats surviving 1 wk, bursting pressure of the anastomotic site was measured as an index of wound healing. At plasma corticosterone levels of 3.9-7.4 micrograms/dl, which approximate normal physiological levels, bursting pressure was not significantly different from that in sham-adrenalectomized animals. In adrenalectomized rats with lower or higher corticosterone levels, bursting pressure was significantly reduced. Thus a narrow range of plasma corticosterone is required for optimal wound healing in this model; higher values tend to impair healing, as do subnormal values, perhaps because of slower protein turnover; low values also lead to high mortality.

Effect of corticosterone administration at varying levels on leucine oxidation and whole body protein synthesis and breakdown in adrenalectomized rats

Adrenalectomized rats were implanted with pellets containing corticosterone in proportions varying from 0% to 100%, plus cholesterol. Stable levels of plasma corticosterone resulted, varying from close to zero (I) to physiologic (II) to supraphysiologic (III). Whole body protein synthesis (S) was measured during fasting by a technique involving injection of [1- 14C]-leucine, analysis of expired air for 14CO 2, and measurement of urinary excretion of urea N plus ammonia N(C). Whole body breakdown (B) during fasting was calculated as the sum of S and C. C rose progressively from I to III, but fractional oxidation of 14C leucine was lowest in II. Both S and B were markedly reduced in I and maximal in II. In III, S fell but B remained high. Thus variations in glucocorticoid levels above and below physiologic exert profound effects on leucine oxidation and protein turnover, involving effects on both synthesis and breakdown.

The role of the adrenal in generating the diurnal variation in circulating levels of corticosteroid-binding globulin in the rat.

Corticosteroid-binding globulin (CBG) levels were measured in serum samples collected sequentially from rats into which indwelling catheters had been inserted. A distinct diurnal variation in CBG levels was found, with the highest levels of binding protein at the beginning of the dark period. CBG levels then decreased until a nadir was reached shortly after the beginning of the light period. To examine the role of glucocorticoid in the generation of this pattern, rats were adrenalectomized 10 days before repeating the experiment. Adrenalectomy abolished both the diurnal variation in CBG levels and the high degree of variation in the levels of binding protein between animals. However, adrenalectomy just before the onset of the dark period did not eliminate the expected decrease in CBG levels. To further explore the role of steroid, animals that had been adrenalectomized for 10 days were given saline containing 25 micrograms/ml corticosterone for a period of 24 h. Despite the attainment of normal plasma corticosterone levels, no decrease in CBG levels was observed. When steroid administration was discontinued, however, CBG levels dropped concurrently with the decreasing steroid. These studies show that a diurnal variation in CBG exists and suggest that it is the result of the diurnal variation in glucocorticoid levels.

Fibrinogen synthesis in serum-free hepatocyte cultures: Stimulation by glucocorticoids

Fibrinogen synthesis was investigated in cultures of chicken embryo hepatocytes initiated and maintained in chemically defined, serum-free medium. 11-Hydroxy glucocorticoids caused a 3-fold stimulation of fibrinogen synthesis. Half-maximal stimulation was achieved with 1 nM corticosterone or hydrocortisone, as compared with only 0.1 nM dexamethasone. Increased fibrinogen production in the presence of these glucocorticoids was characterized by a 4-hr delay in onset, a sensitivity to actinomycin D, and a requirement for the continuous presence of the steroid. Crossed immunoelectrophoresis permitted analysis of the simultaneous effects of glucocorticoids on the synthesis of more than 20 plasma proteins secreted in culture. The absence of an effect on the synthesis of most of these proteins was in sharp contrast to the 3-fold increase in fibrinogen production. Sera from a variety of animals also stimulated an increase in fibrinogen synthesis that was similar in degree but less specific than that due to glucocorticoids and that partially masked the response of the cells to the steroid hormones. The presence of an anticoagulant in the medium was found to be necessary for detection of the fibrinogen secreted in culture. Although insulin was routinely included in the chemically defined medium, the cells synthesized fibrinogen and responded to glucocorticoids in the absence of hormonal supplementation of the medium. These findings are consistent with the thesis that variations in glucocorticoid levels contribute to the regulation of fibrinogen production in the intact animal.