Maternal basking behaviour determines offspring sex in a viviparous reptile

Two primary dichotomies within vertebrate life histories involve reproductive mode (oviparity versus viviparity) and sex determination (genotypic sex determination versus environmental sex determination). Although reptiles show multiple evolutionary transitions in both parameters, the co-occurrence of viviparity and environmental-dependent sex determination have heretofore been regarded as incompatible. Our studies on the viviparous lizard Niveoscincus ocellatus show that the extent of basking by a female influences the sex of her offspring. Critically, our data reveal this effect both in the field (via correlations between date of birth and litter sex ratio) and in a laboratory experiment (females with reduced basking opportunities produced more male offspring). Changes in thermoregulatory behaviour thus allow pregnant female lizards to modify the sex of their offspring.     

Follicular fluid steroid levels and subsequent sex of bovine embryos

On the basis of evidence suggesting a maternal involvement in the determination of the sex of the offspring, we took ova at the point of ovulation from crossbred heifers, fertilised them, and established the sex of the embryos. At the same time we took individual-matched samples of follicular fluid from each follicle of origin, and measured the levels of testosterone and oestradiol, blind to the sex of the embryo. We found no effect of oestradiol on sex in either primary or subordinate follicles. But bovine ova from subordinate follicles that had follicular fluid with a high concentration of testosterone (in vivo) were later more likely to be fertilised by a Y-chromosome-bearing spermatozoon (in vitro). These, along with similar results from other researchers, suggest that further study of the relationship between mammalian follicular hormones at the time of conception and subsequent sex of offspring, may help resolve some of the problems associated with theories of adaptive control of the sex ratio in mammals.     

Genetic control of multiple births in low ovulating mammalian species

In mammals, litter size is a highly variable trait. Some species such as humans or cattle are monotocous, with one or sometimes two newborns per birth, whereas others, the polytocous species such as mice or pigs, are highly prolific and often produce a dozen newborns at each farrowing. In monotocous species, however, two or three newborns per birth may sometime be unwanted. In more polytocous species such as sheep or pigs, litter size is studied in order to increase livestock prolificacy. By contrast, twinning rates in humans or cattle may increase birth difficulties and health problems in the newborns. In this context, the aim of our review was to provide a clearer understanding of the genetic and physiological factors that control multiple births in low-ovulating mammalian species, with particular focus on three species: sheep, cattle, and humans, where knowledge of the ovulation rate in one may enlighten findings in the others. This article therefore reviews the phenotypic and genetic variability observed with respect to ovulation and twinning rates. It then presents the QTL and major genes that have been identified in each species. Finally, we draw a picture of the diversity of the physiological mechanisms underlying multiple ovulation. Although several major genes have been discovered in sheep, QTL detection methods in humans or cattle have suggested that the determinism of litter size is complex and probably involves several genes in order to explain variations in the number of ovulations.     

Genetic control of multiple births in low ovulating mammalian species

In mammals, litter size is a highly variable trait. Some species such as humans or cattle are monotocous, with one or sometimes two newborns per birth, whereas others, the polytocous species such as mice or pigs, are highly prolific and often produce a dozen newborns at each farrowing. In monotocous species, however, two or three newborns per birth may sometime be unwanted. In more polytocous species such as sheep or pigs, litter size is studied in order to increase livestock prolificacy. By contrast, twinning rates in humans or cattle may increase birth difficulties and health problems in the newborns. In this context, the aim of our review was to provide a clearer understanding of the genetic and physiological factors that control multiple births in low-ovulating mammalian species, with particular focus on three species: sheep, cattle, and humans, where knowledge of the ovulation rate in one may enlighten findings in the others. This article therefore reviews the phenotypic and genetic variability observed with respect to ovulation and twinning rates. It then presents the QTL and major genes that have been identified in each species. Finally, we draw a picture of the diversity of the physiological mechanisms underlying multiple ovulation. Although several major genes have been discovered in sheep, QTL detection methods in humans or cattle have suggested that the determinism of litter size is complex and probably involves several genes in order to explain variations in the number of ovulations.

     

Conception date affects litter type and foetal sex ratio in female moose in Estonia

1.  The Trivers–Willard model of optimal sex ratios predicts that in polygynous species mothers in better condition should produce more male than female offspring. However, empirical support for this hypothesis in mammals and especially ungulates has been equivocal. This may be because the fitness of mothers has been defined in different ways, reflecting morphological, physiological or behavioural measures of condition. In addition, factors other than maternal condition can influence a mother's fitness. Given that recent studies of wild ungulates have demonstrated the importance of the timing of conception and birth on offspring fitness, litters conceived at different stages of the rut might be expected to exhibit differences in types and embryonic sex ratio.
2.  Based on a 6-year survey of the reproductive tracts of female moose harvested in Estonia, we investigated the effect of conception date on the types of litters produced and on the foetal sex ratio.
3.  There was a clear relationship between conception date and litter characteristics. Overall, earlier conceived litters were more likely than those conceived late to contain multiple embryos and a high proportion of males. However, while foetal sex ratio varied nonlinearly with conception date in yearlings and subadults, no relationship was found in adults.
4.  We conclude that female moose adjust foetal sex ratio and litter type/size depending on their age and the date of conception, and that these adjustments are in accordance with the Trivers–Willard hypothesis if females that conceive earlier are in better condition.     

Relationship between sex ratio and time of insemination according to both time of ovulation and maturational state of oocyte

The aim of this study was to explore how some reproductive methodologies may affect the sex ratio. We first confirmed the association between the maturation stage of bovine oocytes at the time of in vitro fertilisation (IVF) and the sex ratio of in vitro-derived embryos. Secondly, we studied whether the time of insemination, prior to or after ovulation, could alter the sex ratio in sheep. In the first experiment, bovine oocytes were matured in vitro for 16 h; then oocytes were either fertilised in vitro immediately after extrusion of the first polar body or IVF was delayed for 8 h. The proportion of cleaving embryos and their development to the 8-cell stage was enhanced with delayed insemination. Moreover, delaying IVF produced a male-to-female sex ratio of 1.67:1.00, which was significantly different from the expected 1:1 ratio (p < 0.05), whereas more female embryos were produced when oocytes were fertilised in vitro immediately after polar body extrusion (sex ratio of 1.00:0.67; p < 0.05). In the second experiment, 380 ewes were inseminated at different times before or after ovulation, producing 537 lambs. Significant differences in the sex ratio were obtained when we compared the sex of the offspring of ewes inseminated during the 5 h preceding ovulation (more females) with those inseminated during the 5 h after ovulation (more males). Our results suggest that the differential ability of X- or Y-bearing spermatozoa to fertilise oocytes depending either on time of insemination or oocyte maturation state, may be due, at least partially, to 'intrinsic' differences in the physiological activity of X- or Y-bearing spermatozoa before fertilisation.     

Effects of age and parity on litter size and offspring sex ratio in golden hamsters (Mesocricetus auratus)

Golden hamsters that were mated repeatedly from 55 days of age produced 6-12 litters. Litter size at birth rose between the 1st and 2nd litters, peaked on the 3rd, and declined steadily after the 5th litter. Offspring sex ratio (% male) at birth followed a similar pattern: increasing between the 1st and 2nd litters, remaining high through the 3rd, and becoming increasingly female-biased thereafter. Weaning success decreased sharply after the 6th litter and most dams failed to raise any young to weaning after the 9th litter. These sequential effects on litter size, offspring sex ratio and weaning success were also observed in females mated once at different ages, but they occurred considerably later in life, i.e. increasing parity hastened the effects of advanced age. These age- and parity-related changes in litter composition are consistent with the Trivers-Willard hypothesis that physiologically-stressed females would skew offspring sex ratios to favour daughters. However, since the observed changes in sex ratio were probably due to differential prenatal mortality, their adaptive significance is unclear.     

Litter sex ratios in Richardson’s ground squirrels: long-term data support random sex allocation and homeostasis

When costs of producing male versus female offspring differ, parents may vary allocation of resources between sons and daughters. We tested leading sex-allocation theories using an information-theoretic approach and Bayesian hierarchical models to analyse litter sex ratios (proportion males) at weaning for 1,049 litters over 24 years from a population of Richardson’s ground squirrels (Urocitellus richardsonii), a polygynandrous, annually reproducing mammal in which litter size averages from six to seven offspring and sons are significantly heavier than daughters at birth and weaning. The model representing random Mendelian sex-chromosome assortment fit the data best; a homeostatic model received similar support but other models performed poorly. Embryo resorption was rare, and 5 years of litter data in a second population revealed no differences in litter size or litter sex ratio between birth and weaning, suggesting that litter size and sex ratio are determined in early pregnancy. Sex ratio did not vary with litter size at weaning in any of 29 years, and the observed distribution of sex ratios did not differ significantly from the binomial distribution for any litter size. For 1,580 weaned litters in the two populations, average sex ratio deviated from parity in only 3 of 29 years. Heavier females made a greater reproductive investment than lighter females, weaning larger and heavier litters composed of smaller sons and daughters, but litter sex ratio was positively related to maternal mass in only 2 of 29 years. Such occasional significant patterns emphasize the importance of multi-season studies in distinguishing infrequent events from normal patterns.     

Sex-specific embryonic mortality during concurrent pregnancy and lactation in house mice

To evaluate the importance of lactational stress for sex ratio manipulation in postpartum inseminated house mice, 163 sexually inexperienced male and female BALB/c ABom inbred mice were mated and stud males were removed the day following the initial parturition. Randomly chosen dams were allowed to suckle their young for either one or 14 or 21 days. The results showed that two weeks of lactation enhance the incidence of total abortion of the litter conceived postpartum. If lactation continued another week, dams which gave birth to a second litter lost relatively more embryos during uterine development and the percentage of lost embryos was increased by the number of pups suckled. Among these dams, the number of embryos lost in utero correlated positively with viable litter sex ratios (percentage male pups) at second parturition, indicating that lactational stress enhances embryonic mortality and that this additional mortality predominantly affects female embryos.     

根田鼠母体捕食应激效应与Trivers-Willard 模型的关系

本研究了根田鼠母体捕食应激对其子代出生、断乳和成体体重、窝性比及死亡率的作用,检验Trivers—Willard模型的2个前提条件、母体应激激素在母体投资中的作用,以及母体捕食应激效应与该模型的关系。将妊娠根田鼠母体暴露于其捕食艾鼬,母体应激子代的出生和断乳体重均显降低;到成体,雄性体重有此效应,而雌性体重则接近对照。出生窝性比无变化,但成体窝性比向雌性偏斜。不同年龄阶段的死亡率无显变化,但累计死亡率明显增大。处理雄性子代在断乳和成体时的皮质酮含量显增高,而雌性子代则无显变化,从而验证了Trivers-Willard模型的2个前提条件,提出应激母体激素参与母体对子代的投资观点,并认为,母体捕食应激使根田鼠子代成体窝性比向雌性偏斜的生理投资符合进化稳定对策。     

Effect of time of artificial insemination on embryo sex ratio in dairy cattle

The objective of the present study was to examine whether different intervals between insemination and ovulation have an influence on the sex of seven-day-old embryos in dairy cattle. Cows were inseminated once with semen of one of two bulls of proven fertility between 36 h before ovulation and 12 h after ovulation. Time of ovulation was assessed by ultrasound at 4-h intervals. In total, 64 embryos were determined to be male or female. Of these 64 embryos, 51.6% were female. The sex ratio in the various insemination-ovulation intervals (early: between 36 and 20 h before ovulation; intermediate: between 20 and 8 h before ovulation; late: between 8 h before and 12 h after ovulation) did not significantly differ from the expected 1:1 sex ratio (50, 50 and 55% females, respectively). Bull (Bull A and B) and Parity (primiparous and multiparous) had no influence on the expected 1:1 sex ratio either. The number of cell cycles was similar for male and female (P = 0.23) embryos when quality of the embryo (P < 0.0001) was included in the model. The results of this study indicate that, in cattle, the interval between insemination and ovulation does not influence the sex ratio of seven-day-old embryos.     

Sex-specific fitness returns are too weak to select for non-random patterns of sex allocation in a viviparous snake

When environmental conditions exert sex-specific selection on offspring, mothers should benefit from biasing their sex allocation towards the sex with the highest fitness in a given environment. Yet, studies show mixed support for such adaptive strategies in vertebrates, which may be due to mechanistic constraints and/or weak selection on facultative sex allocation. In an attempt to disentangle these alternatives, we quantified sex-specific fitness returns and sex allocation (sex ratio and sex-specific mass at birth) according to maternal factors (body size, age, birth date, and litter size), habitat, and year in a viviparous snake with genotypic sex determination. We used data on 106 litters from 19 years of field survey in two nearby habitats occupied by the meadow viper Vipera ursinii ursinii in south-eastern France. Maternal reproductive investment and habitat quality had no differential effects on the growth and survival of sons and daughters. Sex ratio at birth was balanced despite a slight female-biased mortality before birth. No sexual mass dimorphism between offspring was evident. Sex allocation was almost random apart for a trend towards more male-biased litters as females grew older, which could be explained by an inbreeding avoidance strategy. Thus, a weak selection for facultative sex allocation seems sufficient to explain the almost equal sex allocation in the meadow viper.     

Sex ratio comparisons between nestlings and dead embryos of the Sparrowhawk Accipiter nisus

Sex ratios that differ from unity have been reported for several bird species, but are poorly understood. Skewed sex ratios may originate at ovulation (primary sex ratio) or arise through differential mortality between the sexes (secondary sex ratio). To estimate the primary sex ratio from nestlings is difficult because in some nests not all the offspring can be sexed. Both when including and excluding such nests, there is a risk of overestimating the proportion of the better-surviving sex. Here we sexed dead Sparrowhawk embryos to determine whether unhatched eggs affect primary sex ratio estimates that are based on nestling data. In nests in which embryo mortality occurred, there was up to a 9% discrepancy in the primary sex ratio estimates based on nestlings alone compared to nestlings and dead embryos together. There was no evidence that these differences were based on sex-specific causes of mortality of embryos.     

Viviparity as a constraint on sex-ratio evolution

Abstract In polytocous mammals, the sex ratio during gestation can influence a variety of morphological, physiological, and life-history traits because of steroid leakage between fetuses. Similar phenomena have also recently been described for a viviparous lizard. Some of these effects have important fitness consequences by influencing reproductive success later in life. Thus, biasing the sex ratio toward one sex may lead to a decreased fitness for the other sex, and therefore constrain the evolution of skewed sex ratios. By incorporating effects of sex ratio on offspring fitness in a simple sex-allocation model, I show that, under some circumstances (1) skewed sex ratios are predicted to evolve, and (2) this cost can constrain the evolution of skewed sex ratios.     

Maternal Gestational Cortisol and Testosterone Are Associated with Trade-Offs in Offspring Sex and Number in a Free-Living Rodent (Urocitellus richardsonii)

The adaptive manipulation of offspring sex and number has been of considerable interest to ecologists and evolutionary biologists. The physiological mechanisms that translate maternal condition and environmental cues into adaptive responses in offspring sex and number, however, remain obscure. In mammals, research into the mechanisms responsible for adaptive sex allocation has focused on two major endocrine axes: the hypothalamic pituitary adrenal (HPA) axis and glucocorticoids, and the hypothalamic pituitary gonadal (HPG) axis and sex steroids, particularly testosterone. While stress-induced activation of the HPA axis provides an intuitive model for sex ratio and litter size adjustment, plasma glucocorticoids exist in both bound and free fractions, and may be acting indirectly, for example by affecting plasma glucose levels. Furthermore, in female mammals, activation of the HPA axis stimulates the secretion of adrenal testosterone in addition to glucocorticoids (GCs). To begin to untangle these physiological mechanisms influencing offspring sex and number, we simultaneously examined fecal glucocorticoid metabolites, free and bound plasma cortisol, free testosterone, and plasma glucose concentration during both gestation and lactation in a free-living rodent (Urocitellus richardsonii). We also collected data on offspring sex and litter size from focal females and from a larger study population. Consistent with previous work in this population, we found evidence for a trade-off between offspring sex and number, as well as positive and negative correlations between glucocorticoids and sex ratio and litter size, respectively, during gestation (but not lactation). We also observed a negative relationship between testosterone and litter size during gestation (but not lactation), but no effect of glucose on either sex ratio or litter size. Our findings highlight the importance of binding proteins, cross-talk between endocrine systems, and temporal windows in the regulation of trade-offs in offspring sex and number.     

Food-restricting first generation juvenile female hamsters (Mesocricetus auratus) affects sex ratio and growth of third generation offspring

Female hamsters (Generation 1) were fed ad libitum or were food-restricted to 65-75% of the amount consumed by controls during their first 50 days of life. Subjects were mated at 91 days of age. Their offspring (Generation 2) were fed ad libitum throughout the experiment, and female offspring were also mated at 91 days of age. Generation 3 litters were monitored every fifth day from birth until Day 25 post partum for litter size, sex ratio, and pup weights. Although there were no significant differences in Generation 3 litter sizes at birth, litters descended from food-restricted Generation 1 females (Group R) were significantly smaller on Days 5-25 than litters descended from control Generation 1 females (Group A). Sex ratios remained significantly greater in Group A than in Group R litters from birth to Day 25 but did not vary over time, suggesting similar post-partum mortality rates for both male and female pups. Weights of Generation 3 male and female pups did not vary significantly within treatments at any time. Group A males weighed significantly more than Group R males from birth through Day 25, but weights of Group A and Group R females were always similar. Food restriction early in life may have long-term consequences on sex ratios of subsequent generations in hamsters.     

Sex ratio trajectory in mouse

Skewing of the sex ratio towards males occurs in humans. The possible explanation for excess male births could be a preference for Y-bearing sperm at fertilization and/or selective elimination of female embryos during pregnancy. In this study, we have tested the sex ratio in the preimplantation embryo (2–3 cells stage/closest possible primary sex ratio), the post-implantation embryo (day E7.5), and at birth (secondary sex ratio) on a homogenous (genetic, environmental, and dietary) population of mice to ascertain the biological reason i.e., male preference at fertilization or female elimination during pregnancy or both. Primary sex ratio on early preimplantation embryos (2–3 cells stage) was studied on 598 embryos and secondary sex ratio (at birth) on 721 pups using PCR-based sexing (both X & Y chromosome-specific) besides sex ratio of 80 post-implantation embryos (day E7.5). We have also investigated whether the fat content (high & low) of the diet affects the sex ratio. We observed a skewed sex ratio (more female) in preimplantation embryos (0.436; 95 % CI 0.39, 0.48), and post-implantation embryos (0.462; 95 % CI 0.35, 0.57) but reverse skewing (more male) at birth (0.539; 95 % CI 0.5, 0.58). We also observed that high-fat diet promoted male sex ratio at birth (0.657; 95 % CI 0.57, 0.74) whereas a low-fat diet had the opposite effect (0.46; 95 % CI 0.36, 0.56) but no effect at fertilization (2–3 cells stage embryos). This indicates selective elimination of female embryo and fetus throughout pregnancy in mice, more so with a high-fat diet.     

Litter size and fetal sex ratio adjustment in a highly polytocous species: the wild boar

For species in which reproductive success is more variable inone sex than the other, the Trivers and Willard model (TWM)predicts that females are able to adjust their offspring sexratio. High-quality mothers should provide greater investmentto one sex than the other. Previous tests of the TWM have beeninconsistent, and whether the TWM applies to species with severaloffspring per litter is unclear due to possible trade-offs betweensize, number, and sex of the offspring. Williams' model (WM)accounts for confounding effects of these trade-offs on sexratio variation. Lastly, the "extrinsic modification hypothesis"predicts changes in offspring sex ratio in relation to climaticconditions and population density. Using wild boar as a model,we tested 1) whether the WM fitted observed sex ratio variationand 2) whether sex ratio variations were related to maternalattributes (test of the TWM) and/or to resource availability(test of the extrinsic modification hypothesis). Females adjustedtheir litter size rather than their litter composition, so thatthe WM was not supported. Likewise, changes in resource availabilitydid not influence the fetal sex ratio, so that the extrinsicmodification hypothesis was not supported. The fetal sex ratiowas negatively related to increasing litter size, providingsome support for the TWM. Sex ratio was male biased for littersizes up to 6 and then became female biased in larger litters.Our results provide the first case study showing marked changesin sex ratio in relation to litter size in a large mammal.     

Reproductive Decisions in Female European Ground Squirrels: Factors Affecting Reproductive Output and Maternal Investment

Physiological and behavioural parameters associated with reproductive effort and success were investigated in female European ground squirrels Spermophilus citellus . The proportion of reproductive (lactating) females in the study population was over 90% and was not related to age. Timing of oestrus and ovulation was found to be affected by the female's emergence date and condition. Females with low emergence mass showed delayed oestrus. Differences in ovulation dates were shown to affect reproductive output in terms of litter size and sex ratio. Early litters were larger and male biased. X-ray techniques were used to determine intrauterine litter size in individual females. The results indicated that litter size and sex ratio were fixed prenatally. Lactation costs were reflected in the intensity of mass loss and duration of lactation. Mass loss varied with litter size, in that females with large litters showed a more rapid loss than others. The second parental investment parameter, lactation duration, varied among individual females and was dependent on the timing of reproduction and litter size (except yearlings). Early born litters, which were, in most cases, larger than later ones, were nursed longer. Prolonged lactation periods affected female condition in that they started prehibernation fattening later and entered hibernation with a lower mass than individuals that had shorter lactation periods. Yearling females probably could not afford the energetic costs of long lactation, independent of their offspring number. These results indicated that females with higher reproductive output and higher investment were unable to compensate these costs before hibernation. Consequences for these individuals could therefore be lower over-winter survival or a delayed oestrus in the following season.     

Male‐specific mortality biases secondary sex ratio in Eurasian tree sparrows Passer montanus

Sex allocation theory predicts that parents bias the offspring sex ratio strategically. In avian species, the offspring sex ratio can be biased at multiple growth stages, although the mechanisms are not well known. It is crucial to reveal a cause and timing of biased offspring sex ratio. We investigated (i) offspring sex ratio at multiple growth stages, from laying to fledging; and (ii) the stage at which offspring sex ratio became biased; and (iii) the cause of biased offspring sex ratio in Eurasian tree sparrows Passer montanus. Sex determination of 218 offspring, including hatchlings and unhatched eggs from 41 clutches, suggested that the offspring sex ratio was not biased at the egg‐laying stage but was significantly female‐biased after the laying stage due to higher mortality of male embryos. Half of the unhatched eggs showed no sign of embryo development (37/74, 50.00%), and most undeveloped eggs were male (36/37, 97.30%). Additional experiments using an incubator suggested that the cause of embryo developmental failure was a lack of developmental ability within the egg, rather than a failure of incubation. This study highlights the importance of clarifying offspring sex ratio at multiple stages and suggests that offspring sex ratio is adjusted after fertilization.