Yolk Hormones Research Articles

Population divergence in maternal investment and embryo energy use and allocation suggests adaptive responses to cool climates.

The thermal sensitivity of early life stages can play a fundamental role in constraining species distributions. For egg-laying ectotherms, cool temperatures often extend development time and exacerbate developmental energy cost. Despite these costs, egg laying is still observed at high latitudes and altitudes. How embryos overcome the developmental constraints posed by cool climates is crucial knowledge for explaining the persistence of oviparous species in such environments and for understanding thermal adaptation more broadly. Here, we studied maternal investment and embryo energy use and allocation in wall lizards spanning altitudinal regions, as potential mechanisms that enable successful development to hatching in cool climates. Specifically, we compared population-level differences in (1) investment from mothers (egg mass, embryo retention and thyroid yolk hormone concentration), (2) embryo energy expenditure during development, and (3) embryo energy allocation from yolk towards tissue. We found evidence that energy expenditure was greater under cool compared with warm incubation temperatures. Females from relatively cool regions did not compensate for this energetic cost of development by producing larger eggs or increasing thyroid hormone concentration in yolk. Instead, embryos from the high-altitude region used less energy to complete development, that is, they developed faster without a concomitant increase in metabolic rate, compared with those from the low-altitude region. Embryos from high altitudes also allocated relatively more energy towards tissue production, hatching with lower residual yolk: tissue ratios than low-altitude region embryos. These results are consistent with local adaptation to cool climate and suggest that this is underpinned by mechanisms that regulate embryonic utilisation of yolk reserves and its allocation towards tissue, rather than shifts in maternal investment of yolk content or composition.

Female red-winged blackbirds (Agelaius phoeniceus) do not alter nest site selection, maternal programming, or hormone-mediated maternal effects in response to perceived nest predation or brood parasitism risk

Predation or brood parasitism risks can change the behaviors and reproductive decisions in many parental animals. For oviparous species, mothers can mitigate their reproductive success in at least three ways: (1) by avoiding nest sites with high predation or parasitism risks, (2) through hormonal maternal effects that developmentally prime offspring for survival in risky environments, or (3) by investing less in reproduction when predation or parasitism risks are high. Here, we tested if perceived predation and parasitism risks can induce any of these behavioral or physiological responses by exposing female red-winged blackbirds (Agelaius phoeniceus) to playbacks of two major nest threats, a predator (Cooper’s hawk, Accipiter cooperii) and an obligate brood parasite (brown-headed cowbird; Molothrus ater), as well as two controls (harmless Eastern meadowlark, Sturnella magna; and silence). We found that female blackbirds did not avoid nesting at sites treated with predator or brood parasite playbacks, nor were females more likely to abandon nesting attempts at these sites. Egg size and yolk hormone profiles, which are common proxies for maternal investment in oviparous species, were statistically similar across treatment sites. Instead, we found intraclutch variation in yolk steroid hormone profiles: concentrations of three progestogens (pregnanedione, 17α-hydroxypregnenolone, and deoxycorticosterone) and two androgens (testosterone and androstenedione) were higher in third-laid than first-laid eggs. Our study largely confirms previous findings of consistent intraclutch yolk hormone variation in this species, in birds in general, and in other oviparous lineages, but uniquely reports on several yolk steroid hormones largely overlooked in the literature on hormone-mediated maternal effects.

Does contaminant exposure disrupt maternal hormones deposition? A study on per- and polyfluoroalkyl substances in an Arctic seabird

Maternal effects are thought to be essential tools for females to modulate offspring development. The selective deposition of avian maternal hormones could therefore allow females to strategically adjust the phenotype of their offspring to the environmental situation encountered. However, at the time of egg formation, several contaminants are also transferred to the egg, including per- and polyfluoroalkyl substances (PFAS) which are ubiquitous organic contaminants with endocrine disrupting properties. It is, however, unknown if they can disrupt maternal hormone deposition. In this study we explored relationships between female PFAS burden and maternal deposition in the eggs of steroids (dihydrotestosterone, androstenedione and testosterone), glucocorticoids (corticosterone) and thyroid hormones (triiodothyronine and thyroxine) in a population of the Arctic-breeding black-legged kittiwake (Rissa tridactyla). Egg yolk hormone levels were unrelated to female hormone plasma levels. Second-laid eggs had significantly lower concentrations of androstenedione than first-laid eggs. Triiodothyronine yolk levels were decreasing with increasing egg mass but increasing with increasing females' body condition. Testosterone was the only transferred yolk hormone correlated to maternal PFAS burden: specifically, we found a positive correlation between testosterone in yolks and circulating maternal perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDcA) and perfluoroundecanoic acid (PFUnA) in first-laid eggs. This correlative study provides a first insight into the potential of some long-chain perfluoroalkyl acids to disrupt maternal hormones deposition in eggs and raises the question about the consequences of increased testosterone deposition on the developing embryo.

Yolk corticosterone and progesterone levels in Greater Rhea (Rhea americana) eggs vary in a changing social environment.

Maternal hormones in avian egg yolks may signal and prepare offspring for the prevailing conditions. However, this adjustment requires some degree of flexibility in regulating yolk hormone deposition. The Greater Rhea (Rhea americana) has a particular mating system that combines mixed polygyny and polyandry, communal nesting, and exclusive paternal care of chicks. In this species, we previously found that yolk hormone deposition varies among eggs of different captive populations and could influence chicks' physiology and behavior. However, it is still unknown whether females can modify yolk hormone deposition in a changing social environment. Using a captive population of Greater Rheas, in this study, we quantified yolk hormone levels before and after a reduction in the number of females present in the population. We found that females deposited on average higher yolk corticosterone and lower yolk progesterone after the change in their social environment. Since corticosterone deposited into the yolk comes exclusively from the female's plasma, our results suggest that females had, on average, higher plasma corticosterone levels. The change in the number of females may increase the events of male-male competitions, courtships, and matings, leading to an increase of corticosterone in the females' plasma and then into their eggs. Since we previously found that higher yolk corticosterone and lower yolk progesterone were associated with the production of chicks that have an attenuated stress response, the present study results suggest that yolk hormone deposition is mediated by flexible mechanisms that could adjust development to the prevailing conditions.

Seasonal changes in yolk hormone concentrations carry-over to offspring traits

Yolk hormones are substances which transmit non-genetic factors from the mother to the next generation. The systematic changes of yolk hormone concentrations within asynchronously hatching clutches have been interpreted as a means to adaptively shape the offspring’s phenotype. However, in synchronously hatching clutches the role of yolk hormones is less understood. We investigated whether seasonal changes between eggs in the yolk hormones testosterone (Testo), progesterone (Prog) and corticosterone (Cort) also occur in the grey partridge, a synchronously hatching precocial species without direct food competition between siblings. Specifically we asked whether yolk hormone concentrations systematically vary with season and whether they affect the offspring’s hatching mass, mass gain, circulating baseline and stress-induced Cort. Additionally, we investigated the effect of genetic background and food availability on yolk hormone concentrations by subjecting grey partridge hens of two strains (wild and domesticated) to two different feeding regimes (predictable vs. unpredictable feeding) during egg laying. We hypothesized that egg hormone concentrations change over the season, but breeding in captivity over many generations and ad libitum food access could have resulted in domestication effects which abolished potential seasonal effects. Results showed that progressing season had a strong positive effect on yolk Prog and yolk Testo, but not on yolk Cort. Feeding regimes and strain had no effect on yolk hormones. Offspring mass and mass gain increased and baseline Cort decreased with progressing season. In addition, yolk Testo correlated positively with offspring mass gain and negatively with baseline Cort, while yolk Prog had a positive correlation with baseline Cort. Strain and feeding regimes of the mother had no effect on offspring traits. In conclusion, grey partridge chicks hatching late in the season might benefit from the increased concentrations of the growth-stimulating yolk Testo and by this catch-up in development. Hence, yolk hormone concentration could adaptively shape the offspring phenotype in a precocial species.

The relationship between male social status, ejaculate and circulating testosterone concentration and female yolk androgen transfer in red junglefowl (Gallus gallus)

Several studies show that avian females prefer males based on their secondary sexual ornaments and dominance status. We tested in red junglefowl (Gallus gallus) how comb size affected the result of fighting and how the dominance status related to testosterone concentrations in their circulation and ejaculates. We subsequently tested how social status was related to female reproductive investment, including yolk hormone transfer. We found that after a fight 1) winners increased plasma T and decreased ejaculates T whereas losers' T remained unchanged, and 2) plasma T of winners was higher but ejaculates T was lower than those of losers. We argued those are consistent with the different reproductive strategies of dominant and subordinate males. Furthermore, in line with offspring sex-dependent growth patterns females transferred significantly more androstenedione to female than male embryos when mated with winners, while doing the opposite when mated with losers. We concluded therefore that female reproductive investment was affected by both partner quality and embryo sex. The results indicate that male quality influences sex-specific maternal investment, which could be mediated by ejaculate testosterone concentration.

Maternal investment in last-laid eggs does not compensate for hatching asynchrony in a seabird.

Maternal and environmental effects can have profound effects on offspring performance by generating variation in offspring phenotypes, independent of genetic effects. Within avian broods, differential maternal investment of resources across the laying sequence is thought to be an adaptive strategy to modulate competitive hierarchies induced by hatching asynchrony. In this study, we evaluated the relative importance of maternally derived within-clutch variation and the asymmetric post-hatching environment for growth and survival of common tern (Sterna hirundo) siblings. We experimentally manipulated hatching order, resulting in chicks from last-laid eggs hatching first and vice versa. Although both initial age and size asymmetries were larger within experimental than control broods, the early survival of last-hatched chicks was similar between groups. Initial positive effects of egg size disappeared as siblings approached fledging. Ultimately at fledging, both within-brood growth and cumulative survival patterns were similar between experimental and control broods, suggesting that the effects of systematic variation of egg constituents (e.g., maternally derived yolk hormones) and egg size are too subtle and largely overwhelmed by the effects of hatching asynchrony. Therefore, we conclude that variation in offspring phenotypes is pre-dominantly determined by the social environment experienced post-hatching. Maternal effects may further fine-tune phenotypic variation in response to varying environmental conditions, but this needs to be tested through empirical studies in which multiple maternal effects are measured simultaneously under different environmental conditions.

Human behaviour at the origin of maternal effects on offspring behaviour in laying hens (Gallus gallus domesticus)

Regular visual presence of humans is known to reduce chickens' human-generated stress responses. Here we questioned whether, more than mere visual presence, human behaviour affects laying hen behaviour and subsequently their offspring's behaviour. We hypothesized that human behaviour triggers maternal effects via variations in yolk hormone levels. For five consecutive weeks, two groups of hens were exposed to the same durations of human presence (30 min twice a day, five days a week) but the behaviour of the human differed between groups. The first group (H+) was exposed to predictable arrival of the experimenter, slow movements combined with static presence, stroking during handling and human voice. Whereas the second group of hens (H-) was exposed to unpredictable arrival of the experimenter which remained silent, in motion, and did not provide stroking during handling. At the end of the treatment, we evaluated egg quality and offspring behaviour. We found that avoidance of the experimenter by H+ hens but not by H– hens decreased significantly. Fertility rates and concentrations of yolk progesterone and estradiol in H+ hens' eggs were higher than in H– hens' eggs. Fear of humans, neophobia or the capacity to solve a detour task did not differ significantly between H+ and H– chicks. Social discrimination tests showed that H+ chicks but not H– chicks typically preferred a familiar conspecific to a stranger. These results show that, with the same duration in the presence of the birds, humans through their behaviour engender variations in fertility rates, yolk hormone levels and transgenerational effects on social skills. Rarely explored, our data suggest that maternal effects influence filial imprinting. These data have broad implications for laboratory, commercial systems and conservatory programs where the inevitable presence of humans could trigger maternal effects on offspring phenotype.

Avian yolk androgens are metabolized rather than taken up by the embryo during the first days of incubation.

Several studies show effects of yolk androgens in avian eggs on the phenotype of the offspring. Yolk hormone concentrations decline strongly within the first few days of incubation. Although early embryonic uptake of yolk androgens is suggested by the presence of radioactivity in the embryo when eggs are injected with radiolabelled androgens, these studies do not verify the chemical identity of radioactive compound(s), although it is known that these androgens can be metabolized substantially. By using stable isotope-labelled testosterone and androstenedione in combination with mass spectrometry, enabling verification of the exact molecular identity of labelled compounds in the embryo, we found that after 5 days of incubation the androgens were not taken up by the embryo. However, their concentrations in the entire yolk albumen homogenates declined strongly, even when corrected for dilution by albumen and water. Our results indicate metabolism of maternal androgens, very likely to 5β-androstane-3α,17β-diol, etiocholanolone and their conjugated forms. The results imply that the effects of increased exposure of the embryo to maternal androgens take place either before this early conversion or are mediated by these metabolites with an as yet unknown function, opening new avenues for understanding hormone-mediated maternal effects in vertebrates.

Effect of Yolk Hormones and Egg Weight on the Sex of Hatch Chicks, Hatchability and Embryonic Mortality of Japanese Quail

Effect of Yolk Hormones and Egg Weight on the Sex of Hatch Chicks, Hatchability and Embryonic Mortality of Japanese Quail

Maternal stress alters the phenotype of the mother, her eggs and her offspring in a wild-caught lizard.

While biomedical researchers have long appreciated the influence of maternally derived glucocorticoids (GCs) on offspring phenotype, ecologists have only recently begun exploring its impact in wild animals. Interpreting biomedical findings within an ecological context has posited that maternal stress, mediated by elevations of maternal GCs, may play an adaptive role preparing offspring for a stressful or rigorous environment. Yet, the influence of maternal stress on offspring phenotype has been little studied in wild animals. We experimentally elevated GCs to ecologically relevant levels (mimicking increases in maternal stress hormones following a nonlethal predator encounter, a heat challenge, or a chasing or confinement stressor) in female eastern fence lizards Sceloporus undulatus during gestation. We tested the hypothesis that maternally derived stress hormones themselves are sufficient to alter offspring phenotype. Specifically, we examined the effects of experimentally elevated maternal GCs on fitness-relevant traits of the mother, her eggs and her subsequent offspring. We found that daily maternal GC elevation: (a) increased maternal antipredator behaviours and postlaying glucose levels; (b) had no effect on egg morphology or caloric value, but altered yolk hormone (elevated GC) and nutrient content; and (c) altered offspring phenotype including stress-relevant physiology, morphology and behaviour. These findings reveal that maternally derived GCs alone can alter offspring phenotype in a wild animal, changes that may be mediated via maternal behaviour, and egg hormone and nutrient content. Understanding the ecological consequences of these effects under different environmental conditions will be critical for determining the adaptive significance of elevated maternal GCs for offspring.

Correction: Social environment during egg laying: Changes in plasma hormones with no consequences for yolk hormones or fecundity in female Japanese quail, Coturnix japonica.

[This corrects the article DOI: 10.1371/journal.pone.0176146.].

Maternal egg hormones in the mating context: The effect of pair personality

AbstractAnimal personality traits emerge developmentally from the interaction of genetic and early environmental factors. Maternal hormones, such as androgens (testosterone, T and androstenedione, A4), transferred to embryos and egg yolks may simultaneously organize multiple behavioural and physiological traits. Although previous studies demonstrated an association between the mother's personality and yolk androgen levels, the independent effects of the male partner's personality and pair combination remains unknown.We test this association using an ecological model species for personality research, the great tit (Parus major) using multiple approaches: (1) a wild population, (2) a randomly mated captive population and (3) an experimental study with (dis)assortatively mated pairs from lines selected for fast exploration/boldness or slow exploration/shyness.Egg androgen concentrations were associated with variation in female personality traits, and the experimental data suggested that this is independent of male personality: Experimental females from the slow‐shy line tended to have higher egg T concentrations than females from the fast‐bold line, with no effect of male personality. Shy females from the wild population had higher egg A4 concentration than bold females. However, in the correlative data yolk hormones were linked with male personality, as well as the interaction between female and male traits: Male handling responsiveness correlated negatively with egg A4 concentration in wild birds. In randomly mated birds, pairs that were mated assortatively for personality had lower egg T concentrations than disassortatively mated pairs.Given that egg androgens are known mediators of avian personality, our results suggest that maternal hormones might contribute to the heritability of personality, may be sensitive to the social context of mating, and act as key drivers of individual differences.Aplain language summaryis available for this article.

Social environment during egg laying: Changes in plasma hormones with no consequences for yolk hormones or fecundity in female Japanese quail, Coturnix japonica.

The social environment can have profound effects on an individual’s physiology and behaviour and on the transfer of resources to the next generation, with potential consequences for fecundity and reproduction. However, few studies investigate all of these aspects at once. The present study housed female Japanese quail (Coturnix japonica) in pairs or groups to examine the effects on hormone concentrations in plasma and yolk and on reproductive performance. Circulating levels of androgens (testosterone and 5-α-dihydrotestosterone) and corticosterone were measured in baseline samples and after standardised challenges to assess the responsiveness of the females’ endocrine axes. Effects of the social environment on female fecundity were analysed by measuring egg production, egg mass, fertilization rates, and number of hatched offspring. Counter to expectation, females housed in pairs had higher plasma androgen concentrations and slightly higher corticosterone concentrations than females housed in groups, although the latter was not statistically significant. Pair vs. group housing did not affect the females’ hormonal response to standardised challenges or yolk testosterone levels. In contrast to previous studies, the females’ androgen response to a gonadotropin-releasing hormone challenge was not related to yolk testosterone levels. Non-significant trends emerged for pair-housed females to have higher egg-laying rates and higher fertility, but no differences arose in egg weight or in the number, weight or size of hatchlings. We propose that our unexpected findings are due to differences in the adult sex ratio in our social treatments. In pairs, the male may stimulate female circulating hormone levels more strongly than in groups where effects are diluted due to the presence of several females. Future studies should vary both group size and sex composition to disentangle the significance of sexual, competitive and affiliative social interactions for circulating and yolk hormone levels, and their consequences for subsequent generations.

Lipids in maternal diet influence yolk hormone levels and post-hatch neophobia in the domestic chick.

We assessed whether the ratio of dietary n-6/n-3 polyunsaturated fatty acids (PUFA) during egg formation engenders transgenerational maternal effects in domestic chicks. We analyzed yolk lipid and hormone concentrations, and HPA-axis activity in hens fed a control diet (high n-6/n-3 ratio) or a diet enriched in n-3 PUFAs (low n-6/n-3 ratio) for 6 consecutive weeks. Their chicks were tested for neophobia during the first week of life. We found higher corticosterone metabolites in droppings of hens fed the diet enriched in n-3 and significantly higher concentrations of yolk progesterone, androstenedione, and estradiol in their eggs compared to controls. Chicks of hens fed the n-3 enriched diet showed a lower body mass at hatch than controls and expressed higher neophobia when exposed to a novel object. These results add support to the hypothesis that the nutritional state of female birds produces variation in yolk hormone levels and engender maternal effects.

Distribution and concentration of maternal progesterone in the yolk of Greater Rhea eggs (Rhea americana).

Progesterone is the most concentrated maternal yolk steroid characterized to date in birds; however, no information about it is available in ratite eggs. We collected freshly laid eggs from zoo-housed Greater Rhea females (Rhea americana) bred under similar rearing conditions during two breeding seasons to characterize concentration and distribution of maternal yolk progesterone. After high-performance liquid chromatography analysis, yolk hormone was measured using a commercial electrochemiluminescence immunoassay. Progesterone concentrations were found to vary significantly among the yolk layers, supporting a follicular origin for this steroid in Greater Rhea eggs. Additionally, highly similar mean absolute yolk progesterone concentrations were detected between 2013 and 2015 breeding seasons (1,332.98 ± 82.59 and 1,313.59 ± 85.19 ng/g, respectively). These values are also comparable to those found in some domestic carinate species. Findings suggest that at population level, when rearing conditions are similar, mean absolute yolk maternal progesterone concentrations also appear bounded. Future research on the factors and mechanisms that regulate progesterone deposition in Greater Rhea eggs is needed to better understand whether its levels depend on different rearing conditions.

Maternal thyroid hormones enhance hatching success but decrease nestling body mass in the rock pigeon (Columba livia)

Thyroid hormones (THs) – triiodothyronine (T3) and thyroxine (T4) – are essential for embryonic development in vertebrates. All vertebrate embryos are exposed to THs from maternal origin. As maternal TH levels are known to be essential to embryonic development, the natural variation of maternal THs probably represents a pathway of maternal effects that can modify offspring phenotype. However, potential fitness consequences of variation of maternal TH exposure within the normal physiological range and without confounding effects of the mother have never been experimentally investigated. We experimentally manipulated the levels of yolk T3 and T4 within the physiological range in a species in which the embryo develops outside the mother’s body, the Rock Pigeon (Columba livia) eggs. Making use of the natural difference of yolk testosterone between the two eggs of pigeon clutches, we were also able to investigate the potential interaction between THs and testosterone. Elevated yolk TH levels enhanced embryonic development and hatching success, and reduced body mass but not tarsus length between day 14 and fledging. The yolk hormones increased plasma T4 concentrations in females but reduced it in males, in line with the effect on metabolic rate at hatching. Plasma concentrations of T3 and testosterone were not significantly affected. The effects of treatment did not differ between eggs with high or low testosterone levels. Our data indicate that natural variation in maternal yolk TH levels affects offspring phenotype and embryonic survival, potentially influencing maternal and chick fitness.

High Yolk Testosterone Transfer Is Associated with an Increased Female Metabolic Rate.

Yolk androgens of maternal origin are important mediators of prenatal maternal effects. Although in many species short-term benefits of exposure to high yolk androgen concentrations for the offspring have been observed, females differ substantially in the amount of androgens they transfer to their eggs. It suggests that costs for the offspring or the mother constrain the evolution of maternal hormone transfer. However, to date, the nature of these costs remains poorly understood. Unlike most previous work that focused on potential costs for the offspring, we here investigated whether high yolk testosterone transfer is associated with metabolic costs (i.e., a higher metabolic rate) for the mother. We show that Japanese quail (Coturnix japonica) females that deposit higher testosterone concentrations into their eggs have a higher resting metabolic rate. Because a higher metabolic rate is often associated with a shorter life span, this relationship may explain the negative association between yolk testosterone transfer and female longevity observed in the wild. Our results suggest that metabolic costs for the mother can balance the short-term benefits of yolk testosterone exposure for the offspring, thereby contributing to the maintenance of variation in maternal yolk hormone transfer in natural populations.

Temperature-induced variation in yolk androgen and thyroid hormone levels in avian eggs

Global warming has substantially changed the environment, but the mechanisms to cope with these changes in animals, including the role of maternal effects, are poorly understood. Maternal effects via hormones deposited in eggs, have important environment-dependent effects on offspring development and fitness: thus females are expected to adjust these hormones to the environment, such as the ambient temperature. Longer-term temperature variation could function as a cue, predicting chick rearing conditions to which yolk hormone levels are adjusted, while short-term temperature variation during egg formation may causally affect hormone transfer to eggs. We studied the effects of ambient temperature on yolk androgens (testosterone and androstenedione) and thyroid hormones (thyroxine and triiodothyronine) in great tits (Parus major) using data from unmanipulated clutches from a wild population and from aviary birds (ad libitum food) exposed to different experimental temperature treatments during five years. Both in the wild and in captivity, longer-term pre-laying ambient temperature was not associated with clutch mean yolk hormone levels, while the way androstenedione and thyroxine levels varied across the laying sequence did associate with pre-laying temperature in the wild. Yolk testosterone levels were positively correlated with short-term temperature (during yolk formation) changes within clutches in both wild and captivity. We also report, for the first time in a wild bird, that yolk thyroxine levels correlated with a key environmental factor: thyroxine levels were negatively correlated with ambient temperature during egg formation. Thus, yolk hormone levels, especially testosterone, seem to be causally affected by ambient temperature. These short-term effects might reflect physiological changes in females with changes in ambient temperature. The adaptive value of the variation with ambient temperatures pre-laying or during egg formation should be studied with hormone manipulations in different thermal environments.

Elevated yolk progesterone moderates prenatal heart rate and postnatal auditory learning in bobwhite quail (Colinus virginianus).

Previous studies have established that yolk hormones of maternal origin can influence physiology and behavior in birds. However, few studies have examined the effects of maternal gestagens, like progesterone, on chick behavior and physiology. We tested the effects of experimentally elevated egg yolk progesterone on embryonic heart rate and postnatal auditory learning in bobwhite quail hatchlings. Quail chicks were passively exposed to an individual maternal assembly call for 10 min/hr during the 24 hr following hatching. Preference for the familiarized call was tested at 48 hr following hatching in three experimental groups: chicks that received artificially elevated yolk progesterone (P) prior to incubation, vehicle-only controls (V), and non-manipulated controls (C). Resting heart rate of P, V, and C embryos were also measured on prenatal day 17. The resting heart rate of P embryos was significantly higher than both the V and C embryos. Chicks from the P group also showed an enhanced preference for the familiarized bobwhite maternal call when compared to chicks from the C and V groups. Our results indicate that elevated yolk progesterone in pre-incubated bobwhite quail eggs can influence arousal level in bobwhite embryos and postnatal perceptual learning in bobwhite neonates.