Elevated Yolk Testosterone Research Articles

Maternal androgens in avian brood parasites and their hosts: Responses to parasitism and competition?

In the coevolutionary dynamic of avian brood parasites and their hosts, maternal (or transgenerational) effects have rarely been investigated. We examined the potential role of elevated yolk testosterone in eggs of the principal brood parasite in North America, the brown-headed cowbird, and three of its frequent host species. Elevated maternal androgens in eggs are a common maternal effect observed in many avian species when breeding conditions are unfavorable. These steroids accelerate embryo development, shorten incubation period, increase nestling growth rate, and enhance begging vigor, all traits that can increase the survival of offspring. We hypothesized that elevated maternal androgens in host eggs are a defense against brood parasitism. Our second hypothesis was that elevated maternal androgens in cowbird eggs are a defense against intra-specific competition. For host species, we found that elevated yolk testosterone was correlated with parasitized nests of small species, those whose nest success is most reduced by cowbird parasitism. For cowbirds, we found that elevated yolk testosterone was correlated with eggs in multiply-parasitized nests, which indicate intra-specific competition for nests due to high cowbird density. We propose experimental work to further examine the use of maternal effects by cowbirds and their hosts.

Side-specific effect of yolk testosterone elevation on second-to-fourth digit ratio in a wild passerine.

Second-to-fourth digit ratio is a widely investigated sexually dimorphic morphological trait in human studies and could reliably indicate the prenatal steroid environment. Conducting manipulative experiments to test this hypothesis comes up against ethical limits in humans. However, oviparous tetrapods may be excellent models to experimentally investigate the effects of prenatal steroids on offspring second-to-fourth digit ratio. In this field study, we injected collared flycatcher (Ficedula albicollis) eggs with physiological doses of testosterone. Fledglings from eggs with elevated yolk testosterone, regardless of their sex, had longer second digits on their left feet than controls, while the fourth digit did not differ between groups. Therefore, second-to-fourth digit ratio was higher in the testosterone-injected group, but only on the left foot. This is the first study which shows experimentally that early testosterone exposure can affect second-to-fourth digit ratio in a wild population of a passerine bird.

Consequences of experimentally elevated yolk testosterone levels for intra- and inter-sexual selection in canaries

Hormones of maternal origin transferred to the eggs of oviparous species have been shown to significantly affect offspring development. Furthermore, there is now increasing evidence that these effects may last into adulthood. This underlines the persistence of yolk hormone-mediated maternal effects as well as their trans-generational potential as these changes may involve fitness-related traits such as mate choice behaviour, reproductive traits and longevity. Here, we tested the potential of yolk testosterone to affect sexual selection by experimentally increasing the yolk testosterone levels via egg injections. We focused on two central axes of sexual selection, male–male competition for access to a female (intra-sexual selection) and female mate choice behaviour (inter-sexual selection), using canaries (Serinus canaria) as a model species. Neither male agonistic behaviour nor access to the opposite sex, as measured in staged male–male encounters in the presence of a female, were affected by experimentally elevating yolk testosterone levels. We did not find any evidence for effects on female mate choice behaviour either, given the lack of significant effects on mate choice activity, consistency in female mate choice or choosiness. In conclusion, our results indicate that the consequences of yolk testosterone for sexual selection through changes in behavioural traits, which are expressed during pair formation or male–male competition, are probably limited.

Maternal dietary carotenoids mitigate detrimental effects of maternal GnRH on offspring immune function in Japanese quail Coturnix japonica

Maternal resources deposited in eggs can aff ect the development of several off spring phenotypic traits and result in tradeoff s among them. For example, maternal androgens in eggs may be benefi cial to off spring growth and competitive ability, but detrimental to immunocompetence and oxidative stress. In contrast, maternal antioxidants in eggs may be benefi cial if they mitigate oxidative stress and immunosuppressive eff ects of androgens. We investigated possible interactive eff ects of maternal steroids and carotenoids on aspects of off spring physiology and phenotype, by simultaneously manipulating levels of androgens (via gonadotropin-releasing hormone, GnRH-challenges) and carotenoids (via diet supplementation) in captive female Japanese quail Coturnix japonica during egg laying. Carotenoid supplementation of hens, which elevates yolk concentrations of carotenoid and vitamins A and E, enhanced egg hatching success, off spring survival to age 15 d, and size of the bursa of Fabricius in off spring. In contrast, repeated maternal GnRH challenges, which elevated yolk testosterone concentrations, enhanced off spring neonatal size, but negatively aff ected bursa size. However, interaction among the treatments suggests that the positive eff ect of maternal carotenoid supplementation on plasma bactericidal capacity was mediated by maternal GnRH challenges. Chicks originating from carotenoid-supplemented hens were less immunosuppressed than those originating from carotenoid-supplemented GnRH-challenged hens, which were less immunosuppressed than chicks from GnRH-challenged females not supplemented with carotenoids. Females availability of carotenoid enriched diets allows them to enhance the development of off spring immune system via carotenoids and vitamins deposited in egg yolks and off set detrimental eff ects of androgens deposited by GnRHchallenged females.

Phenotypic plasticity in response to breeding density in tree swallows: An adaptive maternal effect?

Territorial animals breeding in high-density environments are more likely to engage in aggressive competition with conspecifics for resources necessary for reproduction. In many avian species, increased competition among breeding females results in increased testosterone concentrations in egg yolks. Generally, elevated yolk testosterone increases nestling growth, competitive behaviors, and bold behavioral traits. However, few studies provide an environmental context with which to examine the potential adaptive benefits of these phenotypic changes. In this study, tree swallow (Tachycineta bicolor) breeding density was altered to modify levels of social competition and yolk testosterone. We measured nestling growth, competitive ability, and breathing rate in response to a stressor using a partial cross-foster design. Females breeding at high-density experienced more aggressive, competitive interactions and their eggs had higher testosterone concentrations. Nestlings that hatched in high-density environments grew faster and displayed more competitive behaviors and a higher breathing rate response to a stressor regardless of post-hatching density. Our study demonstrates that phenotypic plasticity occurs in response to yolk testosterone variation resulting from different breeding densities. These findings suggest that naturally-induced maternal effects prepare offspring for competitive environments, supporting the idea that maternal effects are adaptive.

Effects of Prenatal Testosterone Exposure on Antioxidant Status and Bill Color in Adult Zebra Finches

Permanent offspring modification through maternal hormone transfer is thought to be a tool for mothers to influence life-history trajectories of individual offspring. In birds, yolk hormones influence numerous aspects of the offspring's physiology, including antioxidant status, an important physiological measure that is linked to growth, reproductive effort, and survival. While it is evident that yolk hormones can affect antioxidant status of nestlings, it is not known whether their effect extends beyond the nestling stage. In this study, we use the zebra finch (Taeniopygia guttata) to test experimentally whether exposure to elevated yolk testosterone (T) levels can result in long-term effects on antioxidant status and traits likely to be associated with this measure. Our data show a significant but sex-specific effect with respect to a period from fledging to the age of 7 mo; T males had higher antioxidant status than control males, whereas antioxidant levels did not differ among females and were intermediate compared with the two male groups. Bill color, a trait associated with carotenoids (a specific group of antioxidants) and known to be under the control of circulating levels of T, was not affected by our yolk T manipulation. Bill color (alone or in covariation with egg treatment or sex) did not predict immune responsiveness or antioxidant status. Moreover, there was only weak evidence that antioxidant status predicted the strength of different immune responses. Antioxidant status (in covariation with egg treatment and sex) predicted levels of circulating total antibody levels but did not predict the strength of cell-mediated and humoral immune responses. Our results suggest that yolk T affects antioxidant status independently of these other traits.

Pre- and postnatal environmental effects as potential sources of variation in neophobic behaviour in canaries

Neophobia, the fear of new objects or stimuli, has been shown to be affected by both maternal effects and the environmental conditions experienced during development. However, both pathways have so far only been studied in isolation, even though maternal effects are known to have significant effects on early development. Thus, maternal yolk hormones — an important mediator of maternal effects — may affect neophobia both through direct effects on neophobic behaviour and/or by affecting the early development. Both pathways may even act in concert. We measured the neophobic response and habituation to novel food, as well as the response to a novel object in 8-months-old canaries (Serinus canaria). All birds hatched from eggs with elevated yolk testosterone levels or control-treated eggs, and grew up in distinct experimentally-manipulated positions within the sibling hierarchy, which allowed us to test whether and how the effects of prenatally elevated yolk testosterone levels on neophobic behaviour vary with the environmental conditions experienced post-hatch. Neither the experimentally-manipulated yolk testosterone levels nor the position within the sibling hierarchy had a direct effect on neophobic behaviour. The elevation of the yolk testosterone levels modulated the neophobic behaviour and the habituation to novel situations of juvenile canaries only in interplay with the experimentally manipulated position in the sibling hierarchy and offspring sex. The strongest increase of neophobic behaviour was observed in individuals that hatched from eggs with elevated yolk testosterone content while growing up in a superior position in the sibling hierarchy — and in males. However, these effects varied with the focal explanatory variable. Based on our results and a review of the literature, we conclude that neither prenatal maternal nor early environmental effects post-hatching form the main source of variation of neophobia in itself, but that the effects observed as well as the inconsistency thereof can best be explained by the interaction of different partly unknown pathways.

Negative effects of yolk testosterone and ticks on growth in canaries

Maternal yolk hormones in bird eggs are thought to adjust the offspring to the post-hatching environment. This implies that the effects of maternal yolk hormones should vary with the post-hatching environment, but to date such context-dependency has largely been ignored. We experimentally increased yolk testosterone concentrations in canary eggs and simultaneously manipulated the post-hatching context via an experimental tick-infestation of the chicks. This allows us to evaluate the context-dependency of hormone-mediated maternal effects, as it has previously been shown that ectoparasites alter the maternal yolk androgen deposition. The experimental tick infestation reduced growth in chicks from sham-treated eggs, indicating harmful effects of this ectoparasite in canaries. Chicks from testosterone-treated eggs were not affected in their development by ticks, suggesting lower ectoparasite vulnerability. But this may also be due to the fact that experimentally elevated yolk testosterone levels impaired growth even under parasite-free conditions. This contrasts previous studies, but these studies often manipulated first laid eggs, while we used eggs of subsequent laying positions. Later laid eggs are presumably of lower quality and contain higher yolk testosterone concentrations. Thus, the effects of elevated yolk testosterone on growth may be dose-dependent or vary with the egg quality, suggesting prenatal context-dependency.

GnRH - a Missing Link between Testosterone Concentrations in Yolk and Plasma and Its Intergenerational Effects

Despite the strong interest in hormone-mediated maternal effects two key questions concerning their mechanisms are as yet unanswered: First, whether the deposition of hormones in the egg yolk is coupled with the levels of these hormones in the maternal circulation, and second, whether epigenetic changes as induced by embryonic exposure to maternal yolk hormones impinge on yolk hormone deposition at adulthood. We investigated the responsiveness to gonadotropin-releasing hormone (GnRH) in female canaries whose embryonic exposure to yolk testosterone had been manipulated. This enabled us to study to what extent GnRH interlinks testosterone concentrations in female circulation and egg yolk as well as the intergenerational potential of hormone-mediated maternal effects. As expected, canary females responded to GnRH with a rise in plasma testosterone. The GnRH-responsiveness was positively correlated with the yolk testosterone content. Factors stimulating the release of GnRH will, therefore, lead to an increase of testosterone in both plasma and egg, posing a potential constraint on the yolk hormone deposition due to testosterone related trade-offs within the laying female. Exposure to elevated yolk testosterone levels as embryo reduced the GnRH-responsiveness in adulthood, potentially limiting environmental influences on yolk testosterone deposition, but the concentrations of yolk testosterone itself were not affected.

Long-Lasting Consequences of Elevated Yolk Testosterone for Metabolism in the Zebra Finch

Resting metabolic rate is a common way of quantifying the cost of living in endothermic animals. The trait often makes up a substantial part of an animal's energy budget and can also be related to sustainable peak work rate as well as to daily energy expenditure. Studies have shown that metabolic rates are often heritable, but much of the variation seems to be caused by other factors (e.g., environmental and maternal effects). In a previous study, in ovo exposure to increased levels of testosterone induced metabolic costs early in life. It is, however, unknown whether in ovo androgens also have long-term effects on individual metabolic rates. In this study, we show that experimentally increased levels of in ovo testosterone in zebra finches (Taeniopygia guttata) result in a 7% higher resting metabolic rate when they are adults. This shows that maternally transferred hormones can induce long-term effects on metabolic demands and potentially influence variation in life-history strategies among offspring. Variation in maternal hormone transfer may also explain some of the large interindividual variation observed in metabolic rates.

Eggshell characteristics and yolk composition in the common cuckoo Cuculus canorus: are they adapted to brood parasitism?

The developmental rate of cuckoo embryos and their hatching size is greater than that of host species, which may require more nutrient resources in the egg and more intensive gas exchange during development. In the present study, we compared various egg characteristics of a brood parasite, the common cuckoo Cuculus canorus, and its frequent host, the great reed warbler Acrocephalus arundinaceus. As maternally‐derived testosterone is known to enhance growth rate of embryos and hatchlings, cuckoo eggs are expected to contain higher concentration of testosterone than host eggs. In addition, we expected higher concentration of antioxidants in cuckoo eggs to protect embryos from oxidative stress associated with accelerated growth. Our results showed that cuckoo eggs had thicker shells and higher pore density than great reed warbler eggs. Yolk was significantly heavier in cuckoo eggs and contained higher concentrations of carotenoids and vitamin E, however, yolk androgen and immunoglobulin concentrations were lower in cuckoo eggs as compared to great reed warbler eggs. We also examined whether eggshell colour was associated to egg quality, and detected a positive association between blue‐green chroma and yolk antioxidant concentration in both species, suggesting that eggshell colour reflects the antioxidant investment of the female into the eggs. Our results suggest that cuckoo females increase the size, growth rate and competitive ability of their young by providing them with more nutrients and more dietary antioxidants for embryonic development, and not through elevated yolk testosterone or antibody levels. In addition, increased porosity of cuckoo eggshells may allow embryos to develop more rapidly because of a greater capacity of gas exchange.

Maternal yolk testosterone in canary eggs: toward a better understanding of mechanisms and function

Maternal yolk androgens in avian eggs have been shown to affect numerous offspring traits. These changes in offspring phenotype represent examples of maternal effects and are thought to adjust offspring development to the posthatching environment. When studying the functional consequences of yolk hormones it is, therefore, crucial to manipulate the yolk androgen concentrations as well as the environmental conditions under which the study is performed. However, so far context-dependent effects of maternal yolk hormones have not sufficiently been taken into account, which might contribute to the current level of inconsistency in yolk androgen-mediated effects. We experimentally elevated the yolk testosterone concentrations and manipulated the sibling size hierarchy. We focused on the effects of yolk testosterone on growth and monitored begging behavior and parental feeding preferences in search of the underlying mechanisms of changes in growth. Experimental changes in the yolk testosterone concentrations significantly affected offspring phenotype. However, elevated yolk testosterone concentrations only improved the growth of chicks that were at a competitive disadvantage, whereas it benefited all chicks when placed in staged competition for food as juvenile. This emphasizes the compensatory role of yolk androgens in the context of hatching asynchrony and its context dependency. Enhanced growth did not coincide with intensified begging. Neither males nor females preferentially fed chicks hatching from eggs with elevated yolk testosterone concentrations. Enhanced growth rather resulted from yolk testosterone induced changes in physiology, of which the detailed mechanisms are yet unknown.

Long-lasting consequences of elevated yolk testosterone levels on female reproduction

Maternal yolk androgens in bird eggs represent an important pathway along which offspring phenotype is shaped. Most of the hormone-mediated maternal effects are highly important in the context of sibling competition. However, there is also increasing evidence for long-lasting effects far beyond the nestling period, and these effects may have important consequences on the reproductive success of the offspring. Here, we investigated the effects of experimentally elevated yolk testosterone concentrations on growth and reproduction in female canaries. Elevated yolk testosterone concentrations enhanced the post-natal growth rate, but not the asymptotic mass, and reduced the survival probability. The latter may be a consequence of the higher growth rate, which may have rendered females hatching from testosterone-treated eggs (T-females) more vulnerable to harsh environmental conditions. Adult T-females made a larger investment in their clutch by laying more but not heavier eggs than females hatching from control-treated eggs. Our results suggest that the observed long-lasting effect on clutch size relates to changes in the growth trajectory rather than being a direct consequence of testosterone, since studies manipulating early growth conditions obtained similar results. Clearly, further studies are now required in order to investigate the intriguing relationship between yolk testosterone, elevated growth rates, and clutch size.

Yolk testosterone, postnatal growth and song in male canaries

Avian eggs contain substantial amounts of maternal yolk androgens, which have been shown to modulate offspring phenotype. The first studies on the functional consequences of maternal yolk androgens have focused on early life stages and their role in sibling competition. However, recent longitudinal studies reported long-lasting effects of maternal yolk androgens on offspring phenotype, mostly concerning traits that are sensitive to androgens. This suggests that maternal yolk androgens could play an important role in sexual selection, since the expression of many male sexual characters is testosterone-dependent. Using male canaries as a model, we examined the consequences of an experimental elevation of yolk testosterone concentrations on early development as well as long-lasting effects particularly on song, which is one of the most important sexual characters in male songbirds. Elevated yolk testosterone concentrations inhibited male growth, possibly in interaction with an existent ectoparasite exposure. Males hatched from testosterone-treated eggs (T-males) did not have enhanced competitive skills, in contrast to previous studies. The elevation of yolk testosterone concentrations delayed song development but did not affect adult song phenotype. This is intriguing, as yolk testosterone possibly induced developmental stress, which is known to reduce song quality. We hypothesize that yolk testosterone has either no direct effect on adult song phenotype, or that positive effects are merged by the negative effects of developmental stress. Finally, females mated with T-males invested more in their clutch indicating that females either assess T-males as more attractive (differential allocation hypothesis) or compensated for lower offspring viability (compensation hypothesis).

Consequences of maternal yolk testosterone for offspring development and survival: experimental test in a lizard

SummaryHormone‐mediated maternal effects and developmental plasticity are important sources of phenotypic variation, with potential consequences for trait evolution. Yet our understanding of the importance of maternal hormones for offspring fitness in natural populations is very limited, particularly in non‐avian species.We experimentally elevated yolk testosterone by injection of a physiological dose into eggs of the lizardCtenophorus fordiStorr, to investigate its roles in offspring development, growth and survival.Yolk testosterone did not influence incubation period, basic hatchling morphology or survival under natural conditions. However, there was evidence for increased growth in hatchlings from testosterone‐treated eggs, suggesting that maternal hormones have potential fitness consequences in natural populations.The positive effect of prenatal testosterone exposure on postnatal growth could represent a taxonomically widespread developmental mechanism that has evolved into an adaptive maternal effect in some taxa, but remains deleterious or selectively neutral in others.A broader taxonomic perspective should increase our understanding of the role of physiological constraints in the evolution of endocrine maternal effects.

Consequences of prenatal androgen exposure for the reproductive performance of female pheasants ( Phasianus colchicus )

Maternal hormones in vertebrate eggs can mediate important forms of maternal effects. However, the function of hormone transfer to the eggs is still debated, especially because long-term fitness consequences have been little studied. We investigated the effect of prenatal exposure to physiologically elevated yolk testosterone (T) levels on reproduction of female pheasants (Phasianus colchicus) in captivity. We found that females hatching from T-injected eggs (T-females) had a lower egg-laying rate than controls, and their eggs were more frequently infertile than those laid by control females. There were no effects of prenatal maternal treatment on egg size and yolk T concentration, but eggs carrying a female embryo laid by T-females had smaller yolks than eggs with a male embryo, while there was no sex difference in yolk size among the eggs laid by control females. Progeny sex ratio was unaffected by maternal treatment. These findings suggest that the transfer of high androgen levels to the eggs by the mother is constrained by complex trade-offs between direct effects on her daughters' reproduction and by trans-generational differential consequences on male and female descendants.

Effects of elevated yolk testosterone levels on survival, growth and immunity of male and female yellow-legged gull chicks

Androgen hormones of maternal origin contained in the eggs of avian species are considered to have positive effects on offspring characteristics and performance. However, negative consequences have also been reported, suggesting that mothers may experience a trade-off between beneficial and detrimental effects of egg androgens to offspring fitness. We studied the effects of elevated yolk testosterone (T) concentration on survival, development and phenotype of male and female yellow-legged gull (Larus michahellis) chicks by injecting egg yolks with physiological doses of the hormone. Elevated yolk T resulted in a male-biased post-hatching sex ratio, T-treated clutches producing a greater proportion of males compared to control ones at day 4 post-hatching, likely resulting from a reduction of female embryonic survival, whereas no effect of hormone treatment on hatching success or short-term chick survival was observed. In addition, T depressed post-hatching body mass in both sexes but had no effects on the intensity of the cell-mediated immune response or skeletal growth. No sex differences in egg characteristics or chick phenotype were detected. Time to hatching was not affected by T, but females originating from first laid eggs hatched earlier than males of the same laying order, independently of hormone treatment. However, the implications of sex differences in hatching times are unclear in the study species. Taken together, our results suggest that female yellow-legged gulls may be constrained in transferring androgens to their eggs by negative consequences on the viability of female offspring and growth of chicks of the two sexes.

European starling chicks benefit from high yolk testosterone levels during a drought year

Avian egg yolk contains androgenic hormones, such as testosterone, of maternal origin. Experimental elevation of yolk testosterone levels enhances growth of canary chicks. Success in sibling competition, due to increased begging, is presumed to underlie this growth enhancement, because canary hatchlings from testosterone-treated eggs beg longer in response to vibrational stimuli than controls. Furthermore, experimental elevation of both yolk androstenedione and testosterone increased chick growth and begging in black-headed gulls. We measured daily growth of European starling ( Sturnus vulgaris) chicks hatching from testosterone-treated or vehicle-treated (control) eggs until 14 days of age, and measured begging behavior at hatching and at 5 days of age. A temporary drought caused relatively high levels of early brood reduction for this population; 2- and 3-day-old chicks were most likely to starve. We found that chicks from testosterone-treated eggs were less likely to starve than control chicks, and were heavier on the days when most brood reduction occurred. However, chicks from testosterone-treated eggs begged less than control chicks on the day of hatching, and begged similarly at 5 days of age. Thus, while yolk testosterone did increase growth during periods of (presumably) high competition, increased begging does not appear to mediate this effect. Instead, testosterone may induce more efficient energy use, for example, by decreasing ineffective begging. While our results indicate that elevated yolk testosterone enhances survival, and thus offspring and parental fitness, further evidence regarding the fitness consequences of yolk androgens are vital to understanding their role in avian life history.

Regulation of Yolk-Androgen Concentrations by Plasma Prolactin in the American Kestrel

The concentrations of maternally derived androgens in the yolks of avian eggs vary within and among clutches, but a mechanistic basis for this variation has not been elucidated. We investigated in the American kestrel, Falco sparverius, whether changes in plasma-prolactin concentrations induced by changes in photoperiod and food supply affect yolk-androgen concentrations. Over the course of a photoinduced breeding period in the laboratory, we measured concentrations of plasma immunoreactive prolactin (ir-prolactin) in female kestrels with ad libitum food availability (control) or food availability that was reduced during the early breeding period. In a second laboratory study, we administered via osmotic mini-pumps ovine prolactin (o-prolactin) to females beginning on the day they laid their first egg of a clutch (egg-day 1) to determine the effects of high prolactin concentrations on yolk-androgen concentrations. In both this study and one on free-living kestrels, we quantified changes in yolk-androgen concentrations with date of clutch initiation. Concentrations of ir-prolactin in nonlaying females rose with date, irrespective of food treatment. Egg-day 1 ir-prolactin concentrations were higher in control females laying late during the breeding phase than in those laying early. This increase was absent in food-reduced females. Yolk-androgen concentrations in eggs 3 and 4 but not eggs 1 and 2 of the clutch were higher in clutches initiated late than in clutches initiated early in the breeding phase in both the field and laboratory. o-Prolactin treatment elevated yolk-testosterone but not androstenedione concentrations. These findings suggest that, in American kestrels, seasonal and laying-associated increases in plasma-prolactin concentrations elevate yolk-testosterone concentrations. Food availability and other factors may interact with date to regulate the effects of prolactin on yolk-testosterone deposition.