Carotenoid-based Coloration Research Articles

Captivity affects mitochondrial aerobic respiration and carotenoid metabolism in the house finch (Haemorhous mexicanus).

In many species of animals, red carotenoid-based coloration is produced by metabolizing yellow dietary pigments, and this red ornamentation can be an honest signal of individual quality. However, the physiological basis for associations between organism function and the metabolism of red ornamental carotenoids from yellow dietary carotenoids remains uncertain. A recent hypothesis posits that carotenoid metabolism depends on mitochondrial performance, with diminished red coloration resulting from altered mitochondrial aerobic respiration. To test for an association between mitochondrial respiration and red carotenoids, we held wild-caught, molting male house finches in either small bird cages or large flight cages to create environmental challenges during the period when red ornamental coloration is produced. We predicted that small cages would present a less favorable environment than large flight cages and that captivity itself would decrease both mitochondrial performance and the abundance of red carotenoids compared with free-living birds. We found that captive-held birds circulated fewer red carotenoids, showed increased mitochondrial respiratory rates, and had lower complex II respiratory control ratios - a metric associated with mitochondrial efficiency - compared with free-living birds, though we did not detect a difference in the effects of small cages versus large cages. Among captive individuals, the birds that circulated the highest concentrations of red carotenoids had the highest mitochondrial respiratory control ratio for complex II substrate. These data support the hypothesis that the metabolism of red carotenoid pigments is linked to mitochondrial aerobic respiration in the house finch, but the mechanisms for this association remain to be established.

Avian colouration in a polluted world: a meta-analysis.

Brilliant, diverse colour ornaments of birds were one of the crucial cues that led Darwin to the idea of sexual selection. Although avian colouration plays many functions, including concealment, thermoregulation, or advertisement as a distasteful prey, a quality-signalling role in sexual selection has attracted most research attention. Sexually selected ornaments are thought to be more susceptible to external stressors than naturally selected traits, and as such, they might be used as a test for environmental quality. For this reason, the last two decades have seen numerous studies on the impact of anthropogenic pollution on the expression of various avian colour traits. Herein, we provide the first meta-analytical summary of these results and examine whether there is an interaction between the mechanism of colour production (carotenoid-based, melanin-based and structural) and the type of anthropogenic factor (categorised as heavy metals, persistent organic pollutants, urbanisation, or other). Following the assumption of heightened condition dependence of ornaments under sexual selection, we also expected the magnitude of effect sizes to be higher in males. The overall effect size was close to significance and negative, supporting a general detrimental impact of anthropogenic pollutants on avian colouration. In contrast to expectations, there was no interaction between pollution types and colour-producing mechanisms. Yet there were significant differences in sensitivity between colour-producing mechanisms, with carotenoid-based colouration being the most affected by anthropogenic environmental disturbances. Moreover, we observed no significant tendency towards heightened sensitivity in males. We identified a publication gap on structural colouration, which, compared to pigment-based colouration, remains markedly understudied and should thus be prioritised in future research. Finally, we call for the unification of methods used in colour quantification in ecological research to ensure comparability of results among studies.

UV-induced feather color change reflects its porphyrin content.

Pigmentary coloration is widespread in animals. Its evolutionary and ecological features are often attributed to the property of predominant pigments; therefore, most research has focused on predominant pigments such as carotenoids in carotenoid-based coloration. However, coloration results from predominant pigments and many other minority pigments, and the importance of the latter is overlooked. Here, we focused on porphyrin, an "uncommon" pigment found in bird feathers, and investigated its importance in the context of feather color changes in the barn swallow Hirundo rustica. We found that the "pheomelanin-based coloration" of the barn swallow faded after the irradiation of UV light, and this effect was particularly strong in the feathers of young swallows (nestlings and fledglings, here). We also found that it is not the predominant pigment, pheomelanin, but protoporphyrin IX pigment that showed the same pattern of depigmentation after the irradiation of UV light, particularly in the feathers of young swallows. In fact, the abovementioned age-dependent feather color change was statistically explained by the amount of porphyrin in the feathers. The current study demonstrates that a minority pigment, porphyrin, explains within-season dynamic color change, an ecological feature of feather coloration. The porphyrin-mediated rapid color change would benefit young birds, in which feather coloration affects the parental food allocation during a few weeks before independence, but not later. Future studies should not ignore these minor but essential pigments and their evolutionary and ecological functions.

Diet and between-tissue isotope comparisons reveal different foraging strategies for age and sex of a Saffron Finch (Sicalis flaveola Linnaeus, 1766) population.

Measuring stable isotopes in different tissues offers the opportunity to provide insight into the foraging ecology of a species. This study aimed to assess how diet varies between yellow females, yellow males, and dull individuals of a Saffron Finch (Sicalis flaveola) population. We measured δ13C and δ15N in blood over a year, and in different feathers, to estimate seasonal consistency of resource use for each category. We conducted this study in a private farm in the Central Brazilian savannas. We sampled 195 individuals in seven field samplings between January 2017 and March 2018. The mean blood δ13C values were similar among yellow females, yellow males and dull individuals, indicating that this population of Saffron Finch predominantly accesses similar resources throughout the year, with a predominant C4 signal. Although Saffron Finch is considered a granivorous species, the mean δ15N values found indicate that both adults and juveniles also incorporate in their tissues some invertebrate. The slight isotope-tissue difference between feathers and blood is similar to the reported in previous studies and may reflect tissue-to-tissue discrimination. The isotopic space of yellow males was greater than that of yellow females and dull individuals, indicating greater dietary diversity due to greater inter-individual variation in diet. In Saffron Finch, which delays plumage maturation, competition-driven partitioning of food resources seems essential in driving carotenoid-based plumage coloration between age classes and sexes.

Triphenyltin Influenced Carotenoid-Based Coloration in Coral Reef Fish, Amphiprion ocellaris, by Disrupting Carotenoid Metabolism.

Triphenyltin (TPT), a kind of persistent pollutant, is prevalent in the aquatic environment and could pose a threat to coral reef fish. However, little is known about the toxicity of TPT on coral reef fish, especially regarding the representative characteristics of body coloration. Therefore, this study chose the clownfish (Amphiprion ocellaris) in order to investigate the effects of TPT exposure on its carotenoid-based body coloration under the environmentally relevant concentrations (0, 1, 10 and 100 ng/L). After TPT exposure for 60 d, the carotenoid contents were decreased and histological damage in the liver was found, shown as nuclear pyknosis and shift, lipid deposition and fibrotic tissue hyperplasia. Liver transcriptomic analysis showed that TPT exposure interfered with oxidative phosphorylation and fatty acid metabolism pathways, which related to carotenoids uptake and metabolism. Furthermore, TPT exposure led to oxidative damage in the liver, which is responsible for the changes in the antioxidant capacity of enzymes, including GSH, MDA, POD, CAT and T-SOD. TPT exposure also affected the genes (Scarb1, CD36, Stard3 and Stard5) related to carotenoid absorption and transport, as well as the genes (GstP1 and Bco2) related to carotenoid deposition and decomposition. Taken together, our results demonstrate that TPT influenced carotenoid-based coloration in coral reef fish by disrupting carotenoid metabolism, which complements the ecotoxicological effects and toxic mechanisms of TPT and provides data for the body color biology of coral reef fishes.

Multiple ornaments: trade-offs and redundancy in signalling functions in male and female varied tits (Sittiparus varius)

Abstract Signals conveyed by ornaments can be indicators in the process of sexual selection. Specifically, signals may be superimposed on each other to represent an individual’s quality, convey different information, or offer redundant information. As much of the research in this area has focused on relationships between diverse signals in males, those of females have largely been overlooked. From 2022 to 2023, we tested whether three traits, namely acoustic (song display) and visual ornaments (patch size and carotenoid-based plumage coloration), and morphology (wing and tail length), were interrelated in males and whether visual ornaments and morphology were interrelated in females in a wild population of varied tits (Sittiparus varius). We found that males with brighter and more reddish plumage sang at lower maximum and minimum frequencies, with narrower bandwidths and lower frequency peaks. Similarly, a negative correlation between tail length and carotenoid-based plumage coloration was observed in both sexes. Furthermore, a positive relationship was found between signals within traits. These results highlight the trade-offs in signals between different traits, as well as the consistency within the same traits.

Freeze-thaw pretreatment improves the vacuum freeze-drying efficiency and storage stability of goji berry (Lycium barbarum. L.)

In order to improve the efficiency of vacuum freeze-drying of goji berry, the effects of multiple freeze-thaw pretreatments (FT) on the water distribution, microstructure, drying characteristics and physicochemical properties of the goji berry were studied. FT effectively shortened the vacuum freeze-drying duration by 4–6 h and reduced the energy consumption by 10.0–15.1 g/100 g compared with the traditional alkaline-dewaxing method. Further analysis revealed that FT accelerated the whole drying process, especially the rapid drying stage, due to the decrease in the bound water. In addition, FT effectively preserved the physicochemical properties, including shrinkage and hardness of goji berries after freeze-drying treatment. In particular, the deterioration of carotenoid-based red color was alleviated after 6 months of storage of freeze-dried goji berry, during which the degradation of carotenoids was significantly inhibited. Comprehensive analysis established that two freeze-thaw cycles are more suitable for the production of dried berries in practice.

Uridine Diphosphate Glycosyltransferases (UGTs) Involved in the Carotenoid-Based Body Color Difference between Tetranychus cinnabarinus (Red) and Tetranychus urticae (Green).

It has long been disputed whether Tetranychus cinnabarinus and Tetranychus urticae belong to the same genus, with T. cinnabarinus regarded as a red form of T. urticae. However, it is unclear why T. urticae and T. cinnabarinus have different body colors. Since carotenoids are responsible for the color of many organisms, the carotenoid profiles of T. cinnabarinus and T. urticae were compared by HPLC. There was no difference in carotenoid type, but T. cinnabarinus contained significantly more neoxanthin, astaxanthin, α-carotene, β-carotene, and γ-carotene, which may contribute to the deep red color. The transcriptome sequencing of both species identified 4079 differentially expressed genes (DEGs), of which 12 were related to carotenoid metabolism. RNA interference (RNAi) experiments demonstrated that silencing seven of these DEGs resulted in the different accumulation of carotenoid compounds in T. cinnabarinus and T. urticae. In addition, the body of T. urticae turned yellow after two days of feeding with UGT double-stranded RNAs and β-UGT small interfering RNAs. In conclusion, differences in the carotenoid profiles of T. urticae and T. cinnabarinus may be responsible for the different body colors.

Urbanisation impacts plumage colouration in a songbird across Europe: Evidence from a correlational, experimental and meta-analytical approach.

Urbanisation is accelerating across the globe, transforming landscapes, presenting organisms with novel challenges, shaping phenotypes and impacting fitness. Urban individuals are claimed to have duller carotenoid-based colouration, compared to their non-urban counterparts, the so-called 'urban dullness' phenomenon. However, at the intraspecific level, this generalisation is surprisingly inconsistent and often based on comparisons of single urban/non-urban populations or studies from a limited geographical area. Here, we combine correlational, experimental and meta-analytical data on a common songbird, the great tit Parus major, to investigate carotenoid-based plumage colouration in urban and forest populations across Europe. We find that, as predicted, urban individuals are paler than forest individuals, although there are large population-specific differences in the magnitude of the urban-forest contrast in colouration. Using one focal region (Malmö, Sweden), we reveal population-specific processes behind plumage colouration differences, which are unlikely to be the result of genetic or early-life conditions, but instead a consequence of environmental factors acting after fledging. Finally, our meta-analysis indicates that the urban dullness phenomenon is well established in the literature, for great tits, with consistent changes in carotenoid-based plumage traits, particularly carotenoid chroma, in response to anthropogenic disturbances. Overall, our results provide evidence for uniformity in the 'urban dullness' phenomenon but also highlight that the magnitude of the effect on colouration depends on local urban characteristics. Future long-term replicated studies, covering a wider range of species and feeding guilds, will be essential to further our understanding of the eco-evolutionary implications of this phenomenon.

Chromatic and achromatic differences of melanin- and carotenoid-based plumage coloration in five minivet species (Pericrocotus spp.) under conspecific and predator visual systems

Adaptive mate choice has been accepted as the leading theory to explain the colorful plumage of birds. This theory hypothesizes that conspicuous colors act as signals to advertise the qualities of the owners. However, a dilemma arises in that conspicuous colors may not only attract mates, but also alert predators. The “private channels of communication” hypothesis proposes that some intraspecific signals may not be visible to heterospecific animals because of different visual systems. To better understand the evolution of plumage colors and sexual selection in birds, here we studied the chromatic difference and achromatic differences of melanin- and carotenoid-based plumage coloration in five minivet species (Pericrocotus spp.) under conspecific and predator visual systems. We found that either the chromatic or achromatic difference among male or female minivets’ plumage was consistently higher under conspecific vision than under predator vision for all five studied species of minivets. This result indicated that individual differences in plumage colors of minivets were visible to the conspecific receivers and hidden from potential predators as a result of evolution under predation risk and conspecific communication. However, males were under a higher risk of predation because they were more conspicuous than females to the vision of a nocturnal predator.

To colour a bird: The evolution of carotenoid-based colouration in passerines is shaped by sexual selection, ecology and life history.

Research Highlight: Delhey, K., Valcu, M., Dale, J., & Kempenaers, B. (2022). The evolution of carotenoid-based plumage colours in passerine birds. Journal of Animal Ecology, https://doi.org/10.1111/1365-2656.13791. Carotenoids, a class of colour pigments, are responsible for red, yellow and orange hues in nature. They play an important role in visual animals, and specially birds, where dietary carotenoids can act as honest sexual signals. Long-standing interest in the function of carotenoid-based colours has led to different hypotheses for their evolutionary drivers. Yet, comparative studies testing the generality of these hypotheses have been previously limited in phylogenetic scope or resolution. In a recent study, Delhey et al. (2022) combined sexual dichromatism, life history and environmental data to investigate the evolution of carotenoid-based colouration in the largest avian radiation, the passerines (Order: Passeriformes). The authors show that the expression of carotenoid-based colours depends on environmental availability, dietary content and body size. They also show that red carotenoids are more often evolutionarily and metabolically derived, and suggest different colours are favoured by natural and sexual selection. These findings shine new light on commonly held hypotheses of carotenoid-colour evolution and contribute to our understanding of how phenotypic diversity evolves.

Melanin- and Carotenoid-Based Coloration of Plumage and the Level of Aggressiveness: The Relationship of These Parameters in the Greenfinch (Chloris chloris, Passeriformes, Fringillidae)

Melanin- and Carotenoid-Based Coloration of Plumage and the Level of Aggressiveness: The Relationship of These Parameters in the Greenfinch (Chloris chloris, Passeriformes, Fringillidae)

Oestradiol reduces female bill colour in a mutually ornamented bird.

Carotenoid-based colour signals can be costly to produce and maintain, and trade-offs between signalling and other fitness traits are expected. In mutually ornamented species, trade-offs with reproduction may be stronger for females than males, because females often dedicate more resources to offspring production, which may lead to plastic investment in colour signals and plastic sexual dichromatism. Oestradiol is a candidate mediator of this trade-off because it regulates reproductive physiology and may also influence the expression of coloration. We tested this hypothesis by giving female common waxbills (Estrilda astrild) either oestradiol (17β-oestradiol) or empty implants during the early breeding season and measured spectral reflectance of carotenoid-based bill coloration weekly for two months. Using a model of avian vision, we found that bill colour in oestradiol-implanted females became less saturated, less red in hue and brighter, compared with control females and with unimplanted males. This resulted in a change in bill sexual dichromatism from imperceptible to perceptible. Results support the hypothesis that female reproductive physiology influences investment in coloration through changes in oestradiol and show a form of female-driven plastic sexual dichromatism. Greater sensitivity of female colour to physiological and/or environmental conditions helps explain why differences in sexual dichromatism among species differing in ecology often evolve owing to changes in female rather than male phenotype.

Integument carotenoid-based colouration reflects contamination to perfluoroalkyl substances, but not mercury, in arctic black-legged kittiwakes

Anthropogenic activities are introducing multiple chemical contaminants into ecosystems that act as stressors for wildlife. Perfluoroalkyl substances (PFAS) and mercury (Hg) are two relevant contaminants that may cause detrimental effects on the fitness of many aquatic organisms. However, there is a lack of information on their impact on the expression of secondary sexual signals that animals use for mate choice. We have explored the correlations between integument carotenoid-based colourations, blood levels of carotenoids, and blood levels of seven PFAS and of total Hg (THg) in 50 adult male black-legged kittiwakes (Rissa tridactyla) from the Norwegian Arctic during the pre-laying period, while controlling for other colouration influencing variables such as testosterone and body condition. Kittiwakes with elevated blood concentrations of PFAS (PFOSlin, PFNA, PFDcA, PFUnA, or PFDoA) had less chromatic but brighter bills, and brighter gape and tongue; PFOSlin was the pollutant with the strongest association with bill colourations. Conversely, plasma testosterone was the only significant correlate of hue and chroma of both gape and tongue, and of hue of the bill. Kittiwakes with higher concentrations of any PFAS, but not of THg, tended to have significantly higher plasma concentrations of the carotenoids astaxanthin, zeaxanthin, lutein, and cryptoxanthin. Our work provides the first correlative evidence that PFAS exposure might interfere with the carotenoid metabolism and the expression of integument carotenoid-based colourations in a free-living bird species. This outcome may be a direct effect of PFAS exposure or be indirectly caused by components of diet that also correlate with elevated PFAS concentrations (e.g., proteins). It also suggests that there might be no additive effect of THg co-exposure with PFAS on the expression of colourations. These results call for further work on the possible interference of PFAS with the expression of colourations used in mate choice.

No evidence that endohelminth parasites cause selection against hybrid orioles across the Baltimore–Bullock’s Oriole hybrid zone

Abstract The Baltimore–Bullock’s Oriole hybrid zone is one of the best-studied avian hybrid zones in North America, yet our understanding of the causes of selection against hybrids remains poor. We examine if endohelminth parasites may cause selection against hybrid orioles but found no evidence for this hypothesis. Of the 139 male orioles we examined, 43 individuals contained endohelminth parasites from at least 1 of these groups: Cestoda, Acanthocephala, or Nematoda. Across the hybrid zone, Baltimore Orioles (Icterus galbula) and Bullock’s Orioles (I. bullockii) differed in their parasite communities, such that Baltimore Orioles frequently contained both Acanthocephala and Cestoda parasites whereas Bullock’s Orioles primarily contained Cestoda parasites. Despite these differences in parasite communities between parental species, the frequency of hybrid orioles with parasites was similar to parentals, suggesting that hybrids were as susceptible to endohelminth parasites as parentals. Using a subset of 99 adult male orioles, we explored how parasites may be associated with the expression of orange carotenoid-based plumage in hybrids and parentals. Associations between carotenoid-based plumage color and parasites were most strongly expressed in Bullock’s Orioles, but patterns were subtle and counterintuitive because individuals with parasites often had more enhanced color measures compared to individuals without parasites. Taken together, these data suggest that endohelminth parasites impose little fitness costs to male orioles on the breeding grounds and likely do not cause selection against hybrids.

The evolution of carotenoid-based plumage colours in passerine birds.

Many birds use carotenoids to colour their plumage yellow to red. Because birds cannot synthesise carotenoids, they need to obtain these pigments from food, although some species metabolise dietary carotenoids (which are often yellow) into derived carotenoids (often red). Here, we study the occurrence of yellow and red carotenoid-based plumage colours in the passerines, the largest bird radiation and quantify the effects of potential ecological and life-history drivers on their evolution. We scored the presence/absence of yellow and red carotenoid-based plumage in nearly 6,000 species and use Bayesian phylogenetic mixed models to assess the effects of carotenoid-availability in diet, primary productivity, body size, habitat and sexual selection. We also test the widespread assumption that red carotenoid-based colours are more likely to be the result of metabolization. Finally, we analyse the pattern of evolutionary transitions between yellow and red carotenoid-based plumage colours to determine whether, as predicted, the evolution of yellow carotenoid-based colours precedes red. We show that, as expected, both colours are more likely to evolve in smaller species and in species with carotenoid-rich diets. Yellow carotenoid-based plumage colours, but not red, are more prevalent in species that inhabit environments with higher primary productivity and closed vegetation. In general, females were more likely to have yellow and males more likely to have red carotenoid-based plumage colours, closely matching the effects of sexual selection. Our analyses also confirm that red carotenoid-based colours are more likely to be metabolised than yellow carotenoid-based colours. Evolutionary gains and losses of yellow and red carotenoid-based plumage colours indicate that red colours evolved more readily in species that already deposited yellow carotenoids, while the reverse was rarely the case. Our study provides evidence for a general, directional evolutionary trend from yellow to red carotenoid-based colours, which are more likely to be the result of metabolization. This may render them potentially better indicators of quality, and thus favoured by sexual selection.

Same Colors for Different Functions: Implications for the Evolution of Carotenoid-Based Ornamentation.

AbstractSexual ornamentation is often assumed to be costly, allowing honest signaling of individual quality, and carotenoid-based colors have been proposed to bear significant costs. If carotenoid-based colors are costly to produce, sexually selected signals should use more concentrated carotenoid pigments and have more saturated color than nonsexual signals, where honesty-guaranteeing costs are not required. We tested this prediction comparing carotenoid-based colors across canaries, goldfinches, and allies because many of these species use yellow plumage as sexual ornamentation but also have yellow rumps that appear to be nonsexual flash marks. Only in the breast, but not the rump, was there an asymmetric codistribution of male and female color saturation, with males similarly or more saturated than females, indicating evolution of breast color by sexual selection. Yellow was not consistently more saturated in the breast than in the rump, and the codistribution of rump and breast color saturation indicated that saturated rumps can persist irrespective of breast color. This challenges the assumption that carotenoid-based colors bear significant costs. The use of carotenoid coloration as sexual signals in this clade may instead be due to social costs, cost-free index mechanisms for signaling quality, and/or socially monogamous species evolving low-cost signals to mostly discriminate against the lowest-quality mates.

Chemical manipulation of mitochondrial function affects metabolism of red carotenoids in a marine copepod (Tigriopus californicus).

The shared-pathway hypothesis offers a cellular explanation for the connection between ketocarotenoid pigmentation and individual quality. Under this hypothesis, ketocarotenoid metabolism shares cellular pathways with mitochondrial oxidative phosphorylation such that red carotenoid-based coloration is inextricably linked mitochondrial function. To test this hypothesis, we exposed Tigriopus californicus copepods to a mitochondrially targeted protonophore, 2,4-dinitrophenol (DNP), to induce proton leak in the inner mitochondrial membranes. We then measured whole-animal metabolic rate and ketocarotenoid accumulation. As observed in prior studies of vertebrates, we observed that DNP treatment of copepods significantly increased respiration and that DNP-treated copepods accumulated more ketocarotenoid than control animals. Moreover, we observed a relationship between ketocarotenoid concentration and metabolic rate, and this association was strongest in DNP-treated copepods. These data support the hypothesis that ketocarotenoid and mitochondrial metabolism are biochemically intertwined. Moreover, these results corroborate observations in vertebrates, perhaps suggesting a fundamental connection between ketocarotenoid pigmentation and mitochondrial function that should be explored further.

Bigger or long-winged male common crossbills exhibit redder carotenoid-based plumage coloration.

Carotenoid-based ornaments are often considered reliable (honest) individual condition signals because their expression implies physiological costs unaffordable for low-quality animals (handicap signals). Recently, it has been suggested that efficient cell respiration is mandatory for producing red ketocarotenoids from dietary yellow carotenoids. This implies that red colorations should be entirely unfalsifiable and independent of expression costs (index signals). In a precedent study, male common crossbills, Loxia curvirostra, showing a red plumage reported higher apparent survival than those showing yellowish-orange colors. The plumage redness in this species is due to ketocarotenoid accumulation in feathers. Here, we correlated the male plumage redness (a 4-level visual score: yellow, patchy, orange, and red) and the body morphology in more than 1,000 adult crossbills captured in 3 Iberian localities to infer the mechanisms responsible for color evolution. A principal component analysis summarized morphometry of 10 variables (beak, wing, tarsus length, etc.). The overall body size (PC1) and the length of flight feathers regarding body size (PC3) showed significant positive relationships with plumage redness. Plumage redness was barely correlated with bill shape measures, suggesting no constraint in acquiring carotenoids from pine cones. However, large body sizes or proportionally long flying feathers could help carotenoid acquisition via social competition or increased foraging ranges. Proportionally longer flight feathers might also be associated with a specific cell respiration profile that would simultaneously favor flying capacities and enzymatic transformations needed for ketocarotenoid synthesis. Such a phenotypic profile would agree with the hypothesis of ketocarotenoid-based colors acting as individual quality index signals.

Gene expression in male and female stickleback from populations with convergent and divergent throat coloration.

Understanding of genetic mechanisms underlying variation in sexual dichromatism remains limited, especially for carotenoid‐based colors. We addressed this knowledge gap in a gene expression study with threespine stickleback. We compared male and female throat tissues across five populations, including two in which female red coloration has evolved convergently. We found that the expression of individual genes, gene ontologies, and coexpression networks associated with red female color within a population differed between California and British Columbia populations, suggesting differences in underlying mechanisms. Comparing females from each of these populations to females from populations dominated by dull females, we again found extensive expression differences. For each population, genes and networks associated with female red color showed the same patterns for males only inconsistently. The functional roles of genes showing correlated expression with female color are unclear within populations, whereas genes highlighted through inter‐population comparisons include some previously suggested to function in carotenoid pathways. Among these, the most consistent patterns involved TTC39B (Tetratricopeptide Repeat Domain 39B), which is within a known red coloration QTL in stickleback and implicated in red coloration in other taxa.