Christopher R. Friesen

Sexual conflict over mating in red-sided garter snakes (Thamnophis sirtalis) as indicated by experimental manipulation of genitalia

Sexual conflict over mating can result in sex-specific morphologies and behaviours that allow each sex to exert control over the outcome of reproduction. Genital traits, in particular, are often directly involved in conflict interactions. Via genital manipulation, we experimentally investigated whether genital traits in red-sided garter snakes influence copulation duration and formation of a copulatory plug. The hemipenes of male red-sided garter snakes have a large basal spine that inserts into the female cloaca during mating. We ablated the spine and found that males were still capable of copulation but copulation duration was much shorter and copulatory plugs were smaller than those produced by intact males. We also anaesthetized the female cloacal region and found that anaesthetized females copulated longer than control females, suggesting that female cloacal and vaginal contractions play a role in controlling copulation duration. Both results, combined with known aspects of the breeding biology of red-sided garter snakes, strongly support the idea that sexual conflict is involved in mating interactions in this species. Our results demonstrate the complex interactions among male and female traits generated by coevolutionary processes in a wild population. Such complexity highlights the importance of simultaneous examination of male and female traits.

Size dependence in non-sperm ejaculate production is reflected in daily energy expenditure and resting metabolic rate

ABSTRACT The non-sperm components of an ejaculate, such as copulatory plugs, can be essential to male reproductive success. But the costs of these ejaculate components are often considered trivial. In polyandrous species, males are predicted to increase energy allocation to the production of non-sperm components, but this allocation is often condition dependent and the energetic costs of their production have never been quantified. Red-sided garter snakes (Thamnophis sirtalis parietalis) are an excellent model with which to quantify the energetic costs of non-sperm components of the ejaculate as they exhibit a dissociated reproductive pattern in which sperm production is temporally disjunct from copulatory plug production, mating and plug deposition. We estimated the daily energy expenditure and resting metabolic rate of males after courtship and mating, and used bomb calorimetry to estimate the energy content of copulatory plugs. We found that both daily energy expenditure and resting metabolic rate were significantly higher in small mating males than in courting males, and a single copulatory plug without sperm constitutes 5–18% of daily energy expenditure. To our knowledge, this is the first study to quantify the energetic expense of size-dependent ejaculate strategies in any species. Highlighted Article: The daily energy expenditure and resting metabolic rate of red-sided garter snakes are significantly higher in small mating males than in courting males, and a single copulatory plug without sperm constitutes 5–18% of daily energy expenditure.

Ageing and the cost of maintaining coloration in the Australian painted dragon

There is now good evidence in several taxa that animal coloration positively reflects an individuals antioxidant capacity. However, even though telomeres, a marker of ageing, are known to be vulnerable to reactive oxygen species (ROS) attacks, no studies have ever assessed whether colour fading reflects the rate of biological ageing in any taxa. Here, we measured colour fading, telomere erosion (a measure of biological ageing) and ROS levels in painted dragons. We show that individuals that were better at maintaining their coloration during the three months of the study suffered a higher cost in terms of telomere erosion, but overall ROS levels measured at the start of the study were not significantly related to colour maintenance and telomere shortening. We therefore suggest that colour maintenance is a costly phenomenon in terms of telomere erosion, and that overall ROS levels do not seem to be a crucial component linking ornamental coloration and telomere erosion in our study system.

Age-related sex differences in body condition and telomere dynamics of red-sided garter snakes

Life-history strategies vary dramatically between the sexes, which may drive divergence in sex-specific senescence and mortality rates. Telomeres are tandem nucleotide repeats that protect the ends of chromosomes from erosion during cell division. Telomeres have been implicated in senescence and mortality because they tend to shorten with stress, growth and age. We investigated age-specific telomere length in female and male red-sided garter snakes, Thamnophis sirtalis parietalis. We hypothesized that age-specific telomere length would differ between males and females given their divergent reproductive strategies. Male garter snakes emerge from hibernation with high levels of corticosterone, which facilitates energy mobilization to fuel mate-searching, courtship and mating behaviours during a two to four week aphagous breeding period at the den site. Conversely, females remain at the dens for only about 4 days and seem to invest more energy in growth and cellular maintenance, as they usually reproduce biennially. As male investment in reproduction involves a yearly bout of physiologically stressful activities, while females prioritize self-maintenance, we predicted male snakes would experience more age-specific telomere loss than females. We investigated this prediction using skeletochronology to determine the ages of individuals and qPCR to determine telomere length in a cross-sectional study. For both sexes, telomere length was positively related to body condition. Telomere length decreased with age in male garter snakes, but remained stable in female snakes. There was no correlation between telomere length and growth in either sex, suggesting that our results are a consequence of divergent selection on life histories of males and females. Different selection on the sexes may be the physiological consequence of the sexual dimorphism and mating system dynamics displayed by this species.

Female behaviour and the interaction of male and female genital traits mediate sperm transfer during mating

Natural selection and post‐copulatory sexual selection, including sexual conflict, contribute to genital diversification. Fundamental first steps in understanding how these processes shape the evolution of specific genital traits are to determine their function experimentally and to understand the interactions between female and male genitalia during copulation. Our experimental manipulations of male and female genitalia in red‐sided garter snakes (Thamnophis sirtalis parietalis) reveal that copulation duration and copulatory plug deposition, as well as total and oviductal/vaginal sperm counts, are influenced by the interaction between male and female genital traits and female behaviour during copulation. By mating females with anesthetized cloacae to males with spine‐ablated hemipenes using a fully factorial design, we identified significant female–male copulatory trait interactions and found that females prevent sperm from entering their oviducts by contracting their vaginal pouch. Furthermore, these muscular contractions limit copulatory plug size, whereas the basal spine of the male hemipene aids in sperm and plug transfer. Our results are consistent with a role of sexual conflict in mating interactions and highlight the evolutionary importance of female resistance to reproductive outcomes.

Evolutionary ecology of telomeres: a review

Telomere‐induced selection could take place if telomere‐associated disease risk shortens reproductive life span and differently reduces relative fitness among individuals. Some of these diseases first appear before reproductive senescence and could thus influence ongoing selection. We ask whether we can estimate the components of the breeders equation for telomeres, in which the response to selection (R, by definition “evolution”) is the product of ongoing selection (S) and heritability (h2). However, telomere inheritance is a conundrum: in quantitative genetics, traits can usually be allocated to categories with relatively high or low heritability, depending on their association with relative fitness. Telomere traits, however, show wide variation in heritability from zero to one, across taxa, gender, ethnicity, age, and disease status. In spite of this, there is divergence in telomere length among populations, supporting past and ongoing telomere evolution. Rates of telomere attrition and elongation vary among taxa with some, but not complete, taxonomic coherence. For example, telomerase is commonly referred to as “restricted to the germ line in mammals,” but inbred mice and beavers have telomerase upregulation in somatic tissue, as do many ectotherms. These observations provoke a simplistic understanding of telomere evolutionary biology—clearly much is yet to be discovered.

Telomere dynamics in a lizard with morph-specific reproductive investment and self-maintenance

Abstract Telomeres in human fibroblasts shorten progressively during in vitro culturing and trigger replicative senescence. Furthermore, shortened telomeres can be used as biomarkers of disease. These observations have led to the suggestion that telomere dynamics may also be associated with viability and selection for life history variation in non‐human taxa. Model systems to examine this suggestion would particularly benefit from the coexistence of multiple phenotypes within the same species with different life history trade‐offs, since those could be compared in terms of telomere characteristics. This scenario also provokes the classic question of why one morph does not have marginally higher fitness and replaces the others. One explanation is that different morphs have different reproductive tactics with equal relative fitness. In Australian painted dragons (Ctenophorus pictus), males differ in head color, the presence or absence of a gular bib, and reproductive expenditure. Red males out‐compete yellow males in dominance contests, while yellow males copulate quickly and have higher success in sperm competition than red males. Males with bibs better defend partners against rival matings, at the cost of loss of body condition. We show that yellow‐headed and bib‐less males have longer telomeres than red, blue and bibbed males, suggesting that telomere length is positively associated with higher investment into self‐maintenance and less reproductive expenditure.

Females remate more frequently when mated with sperm-deficient males

Polyandry is a source of sexual conflict and males often try to limit female promiscuity. Consequently, male manipulation of receptivity via antiaphrodisiacs and copulatory plugs that prevent female remating can be a source of sexual conflict. This sexual conflict may be intensified when females must remate for fertility insurance. Male red-sided garter snakes produce a large, gelatinous copulatory plug that has been proposed to 1) physically prevent remating and 2) contain an antiaphrodisiac that reduces female receptivity. These males may become sperm depleted because of their dissociated reproductive pattern. If a female mates with a sperm deficient male and is also rendered unreceptive to further mating, then this represents a serious conflict. We tested whether female remating frequency is affected when females are mated with a male that produces a sperm-less copulatory plug. We show that females are significantly more likely to remate after mating with vasectomized males than intact males, even though vasectomized males still produce a copulatory plug. These results suggest that the ejaculate material of the plug does not contain an antiaphrodisiac. Instead, females may use sperm as a cue for post-copulatory mate assessment and seek to remate for the direct benefit of fertility insurance if they have mated with sperm-depleted males.

Conditional Handicaps in Exuberant Lizards: Bright Color in Aggressive Males Is Correlated with High Levels of Free Radicals

Two non-independent theories in evolutionary ecology, the immunocompetence and the oxidative handicap hypotheses, may explain maintenance of genetic variation and signal honesty through genic capture in sexual selection biology. Thus, polygenic traits, like body condition, could help maintain variation in signalling traits under strong, directional sexual selection while maintaining signal honesty. The immunocompetence and oxidative handicap hypotheses are complementary in this regard in that hormones (most often steroids) essential for sexual signalling also carry costs, including increases in potentially damaging reactive oxygen species (ROS). Here we combine (at least) two of these hypotheses in a study of morph-specific condition and free radical effects on signalling traits (head coloration) in males of a polymorphic lizard, the Australian painted dragon (Ctenophorus pictus). Males differ in head colour (red, orange, yellow, hereto forth collectively ‘yellow-reds’), with these morphs showing reproductive tactics and behaviour in a graded fashion of aggression and dominance (red highest levels). A blue morph recently emerged in our study population and has never been behaviourally characterised. Body condition varied significantly in its relationship with superoxide among the four morphs, with males in better condition showing higher superoxide levels in ‘yellow-reds’ (least so in red males). Blue males contrasted markedly by showing lower superoxide levels in males in better condition, perhaps facilitated by no (or reduced) dermal deposition of pigmentation. Colour degradation with loss in condition from yellow to red males, suggesting that red males (i.e., genes associated within a more aggressive morph) is more able to maintain colour with superoxide acting as a potential handicap. This result is consistent with handicap principles in that males with the more pronounced signal carry a higher cost (higher ROS levels) when being in better body condition, while maintaining more vivid coloration (the condition-dependent trait).

Experimental heatwaves negatively impact sperm quality in the zebra finch

For sexually reproducing species, functionally competent sperm are critical to reproduction. While high atmospheric temperatures are known to influence the timing of breeding, incubation and reproductive success in birds, the effect of temperature on sperm quality remains largely unexplored. Here, we experimentally investigated the impact of ecologically relevant extreme temperatures on cloacal temperature and sperm morphology and motility in zebra finches Taeniopygia guttata. We periodically sampled males exposed to 30°C or 40°C temperatures daily for 14 consecutive days. Following a 12-day (23°C) recovery period, birds were again exposed to heat, but under the alternate treatment (e.g. birds initially exposed to 40°C were exposed to 30°C). Elevated temperatures led to an increase in cloacal temperature and a reduction in the proportion of sperm with normal morphology; these effects were most notable under 40°C conditions, and were influenced by the duration of heat exposure and prior exposure to high temperature. Our findings highlight the potential role of temperature in determining male fertility in birds, and perhaps also in constraining the timing of avian breeding. Given the increased frequency of heatwaves in a warming world, our results suggest the need for further work on climatic influences on sperm quality and male fertility.