Geographic variation in timekeeping systems among three populations of garter snakes (Thamnophis sirtalis) in a common garden

Transduction of environmental cues into endocrine signals that synchronize physiology and behavior with optimal environmental conditions is central to an animal's timekeeping system. Using a common garden approach, we investigated possible geographic variation in timekeeping systems by comparing 24-h melatonin and corticosterone rhythms and reproductive behavior among three populations of garter snakes with very different life histories: red-sided garter snakes (Thamnophis sirtalis parietalis) from Manitoba, Canada; red-spotted garter snakes (Thamnophis sirtalis concinnus) from western Oregon; and eastern garter snakes (Thamnophis sirtalis sirtalis) from southern Florida. Melatonin and corticosterone cycles differed significantly among the three snake populations in a majority of the sampling periods. Population differences were observed across a wide range of acclimatization conditions and were themselves plastic (i.e., one snake population was not consistently different from the others). Changes in courtship behavior during emergence also varied significantly among populations. Our data support the hypothesis that endogenous timekeeping systems have evolved in the presence of unique environmental conditions. Further research is necessary to determine whether this geographic variation results from inherent genetic differences or whether it is a product of development. These studies provide insight into the evolution of timekeeping systems and may aid in understanding the potential effects of environmental perturbations on seasonal physiology and behavior.     

Patterns of genetic differentiation in Thamnophis and Taricha from the Pacific Northwest

Aim The co‐evolutionary interaction between the common garter snake, Thamnophis sirtalis, and the rough‐skinned newt, Taricha granulosa, takes place throughout much of the Pacific Northwest (North America). The biogeography of the Pacific Northwest has been heavily influenced by the last Pleistocene glaciation, which reached a maximum as late as 14,000 yr bp . We researched: (1) what type of population structure is present for garter snakes and newts, (2) whether the population structure of these species is consistent with a Pleistocene glaciation hypothesis, and (3) how population structure and migration possibly affect co‐evolution between these species. Location The Pacific Northwest of North America, specifically northern California, Oregon and Washington in the USA. Methods We sampled approximately 20 populations for each species from three different transects. Using microsatellite markers and tissue samples from both species, we quantified the population structure for both species. Individual‐based assignment tests were used to estimate contemporary migration rates. Results Both Th. sirtalis and Ta. granulosa exhibited little genetic differentiation among our study sites, even among those separated by large distances. Significant population structure was detected on multiple geographic scales. Differences in population structure were observed among transects and between garter snake and newt transects. Contemporary migration rate estimates indicate high levels of genetic exchange between populations. Main conclusions Prior to this study, little was known about the fine‐scale population structure of either species in this region. Patterns of population structure for garter snakes and newts reflect a shared biogeographical history affected by the Pleistocene glaciation in the Pacific Northwest. Both species apparently migrate frequently between populations, thus potentially retarding the process of adaptive co‐evolution. We find that populations from a northern coastal transect (Washington) are most likely to be locally adapted.     

Hematology and plasma biochemistry values for the giant garter snake (Thamnophis gigas) and valley garter snake (Thamnophis sirtalis fitchi) in the Central Valley of California

Hematology and plasma biochemistry parameters are useful in the assessment and management of threatened and endangered species. Although reference ranges are readily available for many mammalian species, reference ranges for snakes are lacking for most species. We determined hematology and plasma biochemistry reference ranges for giant garter snakes (Thamnophis gigas) and valley garter snakes (Thamnophis sirtalis fitchi) living in four management areas in the Central Valley of California. White blood cell, heterophil, lymphocyte, and azurophil counts in giant garter snakes were approximately twice the values of valley garter snakes. Statistically significant differences in aspartate aminotransferase, globulin, and potassium between the two species did not appear clinically significant. No significant differences were found in the measured parameters between male and female giant garter snakes. Some differences were found among collection sites. These reference ranges provide baseline data for comparisons over time and between collection sites.     

Molecular phylogeography of common garter snakes (Thamnophis sirtalis) in western North America: implications for regional historical forces

Complete ND2 and partial ND4 and cytochrome b mitochondrial DNA (mtDNA) sequences were analysed to evaluate the phylogeographic patterns of common garter snakes (Thamnophis sirtalis) in western North America. This species is widely distributed throughout North America, and exhibits extensive phenotypic variation in the westernmost part of its range. The overall phylogeographic pattern based on mtDNA sequences is concordant with results from studies of other species in this region, implicating historical vicariant processes during the Pleistocene and indicating bottleneck effects of recent dispersal into postglacial habitat. Indeed, the topology is statistically consistent with the hypothesis of both southern (Great Basin and California) and northern (Haida Gwaii) refugia. Specifically, we identified genetic breaks among three major clades: Northwest Coastal populations, Intermountain populations, and all California populations. The California clade contained the only other well-supported branching patterns detected; relationships among populations within the two northern clades were indistinguishable. These molecular splits contrast sharply with all prior geographical analyses of phenotypic variation in T. sirtalis in this region. Our results suggest that the extensive phenotypic variation in western T. sirtalis has been shaped more by local evolutionary forces than by shared common ancestry. Consequently, we consider all morphologically based subspecies designations of T. sirtalis in this region invalid because they do not reflect reciprocal monophyly of the mtDNA sequences.     

The energetic advantages of slug specialization in garter snakes (genus Thamnophis)

We tested the hypothesis that dietary specialization by foraging garter snakes is accompanied by increased assimilation efficiency on specialist prey items. Our comparison included two closely related garter snake species considered to be slug specialists (Thamnophis ordinoides and Thamnophis elegans terrestris), one fish specialist (Thamnophis couchii), and one diet generalist (Thamnophis elegans elegans). Our results suggest that slug specialists have an energetic advantage over non-slug-eating snakes when both eat slugs. Slug specialists T. ordinoides and T. e. terrestris both have higher assimilation and net assimilation efficiencies when eating slugs than do generalists T. e. elegans and T. couchii. The slug specialists did not experience decreased efficiency when eating fish. Therefore, there was no apparent digestive trade-off for the slug specialists when eating other prey.     

Helminth infracommunity structure of the sympatric garter snakes Thamnophis eques and Thamnophis melanogaster from the Mesa Central of Mexico

Seventy-two Mexican garter snakes (Thamnophis eques) and 126 black-bellied garter snakes (T. melanogaster) were collected from 4 localities of the Mesa Central of Mexico between July 1996 and February 1998 and examined for helminths. Both species of garter snakes occurred sympatrically in every locality except in Lake Cuitzeo. Both species of snakes shared 9 helminth species, and in general, T. melanogaster hosted a larger number of species than T. eques. In each locality, a different helminth species showed the highest levels of prevalence and abundance (Spiroxys susanae in Ciénaga de Lerma, Telorchis corti in Lago de Pátzcuaro, Proteocephalus variabilis in Lago de Cuitzeo, and Contracaecum sp. in Lago de Chapala). Helminth communities in garter snakes of the Mesa Central are depauperate and dominated by a single parasite species. In those localities where the snakes occurred in sympatry, helminth communities were, in general, more diverse and species-rich in T. melanogaster. Differences in the ecology and physiology of these species of garter snakes may explain this pattern because black-bellied garter snakes (T. melanogaster) are more aquatic than Mexican garter snakes (T. eques) and primarily eat aquatic prey, potentially exposing themselves to a larger number of helminths transmitted by predator-prey infection. The helminth infracommunities of garter snakes in the Mesa Central of Mexico show a strong Nearctic influence because most of the species infecting these hosts have been recorded in other Nearctic colubrid snakes. However, the helminth infracommunities of these garter snakes are less species-rich and less diverse than those in colubrid snakes in more temperate latitudes. The widespread ecological perturbation of sampling sites in the Mesa Central because of human activity, and geographic differences in foraging ecology of the hosts and, thus, exposure to parasites transmitted by intermediate hosts may help to explain these patterns.     

Species specificity of methyl ketone profiles in the skin lipids of female garter snakes,genus Thamnophis

One of the relatively few vertebrate pheromones to be chemically identified, the female sex pheromone of the red-sided garter snake (Thamnophis sirtalis parietalis) is a series of saturated and monounsaturated methyl ketones contained within female skin lipids. During the breeding season, this pheromone is responsible for eliciting male courtship behaviors and males are able to utilize pheromonal variation to discriminate among females. While the pheromone system of the red-sided garter snake has been the subject of many studies, relatively little is known about the pheromone systems of other garter snakes. Through chemical analyses, we demonstrate that female skin lipids of the red-spotted garter snake (Thamnophis sirtalis concinnus), northwestern garter snake (Thamnophis ordinoides), and plains garter snake (Thamnophis radix) contain similar methyl ketones. The methyl ketone profiles of these snakes differ qualitatively from one another and from the methyl ketone profiles of red-sided garter snakes with differences particularly pronounced between sympatric species. Our results provide evidence that the use of methyl ketones in sexual signaling may be ubiquitous for Thamnophis species and suggest that these compounds could play a role in reproductive isolation between species in this genus.     

Genetic architecture of a feeding adaptation: garter snake (Thamnophis) resistance to tetrodotoxin bearing prey

Detailing the genetic basis of adaptive variation in natural populations is a first step towards understanding the process of adaptive evolution, yet few ecologically relevant traits have been characterized at the genetic level in wild populations. Traits that mediate coevolutionary interactions between species are ideal for studying adaptation because of the intensity of selection and the well-characterized ecological context. We have previously described the ecological context, evolutionary history and partial genetic basis of tetrodotoxin (TTX) resistance in garter snakes (Thamnophis). Derived mutations in a voltage-gated sodium channel gene (Na_v1.4) in three garter snake species are associated with resistance to TTX, the lethal neurotoxin found in their newt prey (Taricha). Here we evaluate the contribution of Na_v1.4 alleles to TTX resistance in two of those species from central coastal California. We measured the phenotypes (TTX resistance) and genotypes (Na_v1.4 and microsatellites) in a local sample of Thamnophis atratus and Thamnophis sirtalis. Allelic variation in Na_v1.4 explains 23 per cent of the variation in TTX resistance in T. atratus while variation in a haphazard sample of the genome (neutral microsatellite markers) shows no association with the phenotype. Similarly, allelic variation in Na_v1.4 correlates almost perfectly with TTX resistance in T. sirtalis, but neutral variation does not. These strong correlations suggest that Na_v1.4 is a major effect locus. The simple genetic architecture of TTX resistance in garter snakes may significantly impact the dynamics of phenotypic coevolution. Fixation of a few alleles of major effect in some garter snake populations may have led to the evolution of extreme phenotypes and an ‘escape’ from the arms race with newts.     

Experimental evidence for the adaptive evolution of growth rate in the garter snake Thamnophis elegans

The western terrestrial garter snake (Thamnophis elegans) varies significantly in individual growth rates and life-history traits (maturation, reproduction, and survival) among adjacent populations in nature. This study focuses on assessing the genetic and environmental components of the substantial among-population variation in growth rates. Litters of neonates from nine populations inhabiting either mountain meadow or lakeshore habitat were reared for one year in a common-garden experiment with two temperature treatments. Diet, frequency of feeding, light exposure, and daytime temperatures were identical for all individuals. The two different nighttime temperatures (20 degrees C and 25 degrees C) were chosen to mirror field differences in nighttime thermoregulatory constraints for mountain-meadow and lakeshore snakes, respectively. Temperature and source habitat interacted to affect first-year growth rate. Neonates from meadow dams grew fastest in the cooler treatment, whereas those from lakeshore dams grew fastest in the warmer treatment. The observation that naive neonates, which were gestated and raised under identical conditions, grew fastest in environments characteristic of their natal population is evidence both that there are genetic differences among populations for growth and that these differences reflect adaptation to local habitats at a very small geographic scale. In addition, significant directional selection for large birthweight was measured for neonates from all populations. These results are considered in the context of population colonization history, migration and selection, and competing models for growth rate variation.     

Gene flow and melanism in Lake Erie garter snake populations

Melanistic garter snakes ( Thamnophis sirtalis ) are unusually common near Lake Erie, apparently because selection for thermoregulatory ability in cool lake-shore habitats (which favours melanistic morphs) outweighs selection for crypsis (which favours striped morphs). However, morph frequencies are highly variable among sites, suggesting that random genetic drift also influences colour pattern. In an effort to better understand the evolutionary processes influencing garter snake colour patterns, we estimated Fx and Nm (the number of migrants per generation) among island and mainland populations from patterns of allozymic variation detected using electrophoresis. Estimates of Nm were high, ranging from 2.7 to 37.6 between pairs of study sites and making it unlikely that differences in morph frequencies among sites were solely the result of random genetic drift. Furthermore, differences in F _st estimates between colour pattern (a one-locus two-allele trait) and allozyme loci suggest that colour pattern alleles are not in Hardy-Weinberg equilibrium, most likely as a result of natural selection. Comparison of allozymic data from Lake Erie with those from more distant sites suggests that gene flow occurs over long distances in T. sirtalis.     

Discordant patterns of population structure for two co‐distributed snake species across a fragmented Ontario landscape

Aim The goal of our study was to investigate the effects of a fragmented landscape on the genetic population structure of two sympatric snake species that differ in habitat preference. The eastern garter snake (Thamnophis sirtalis sirtalis) is a common, habitat generalist, whereas the endangered eastern foxsnake (Mintonius [Elaphe] gloydi) is rarer, geographically restricted, and a marsh‐specialist. We were most interested in comparing the genetic population structure of both species and identifying any natural and human‐created features of the landscape that overlap with genetic disjunctions. Location Southwestern Ontario, Canada, surveying over half of the remaining range of the eastern foxsnake. Methods We utilized DNA microsatellite markers to examine genetic population structure of both species. The number of genetically distinct clusters for each species was determined using both Bayesian spatial assignment and spatial principal component analyses (sPCA). Genetic clusters were overlaid onto a habitat map to deduce possible physiognomic barriers to gene flow. Results Spatial assignment revealed three genetic clusters for garter snakes and five for foxsnakes. Each individual garter snake had a near equal probability of membership to two or more clusters with no cluster mapping onto a discrete geographic region, indicating that garter snakes comprise a single genetic population. The identified foxsnake clusters correspond to geographically circumscribed locations on the landscape, roughly coincident with isolated patches of suitable habitat. sPCAs revealed significant global allelic structure for foxsnakes, but not for garter snakes. No significant local structure was found for either species. Main Conclusions Our results imply that foxsnakes and garter snakes are differentially impacted by the same landscape or have dramatically different effective population sizes. Unsuitable intervening habitat such as agricultural tracts and roads between existing populations of foxsnakes appears to act as barriers to gene flow, while garter snake movement appears unrestricted by these features. Our findings have important implications for the management of eastern foxsnakes.     

Kinematics of swimming garter snakes (Thamnophis sirtalis)

We investigate the kinematics of swimming garter snakes (Thamnophis sirtalis) using a novel nonlinear regression-based digitization method to establish quantitative statistical support for non-constant wavelengths in the undulatory pattern exhibited by swimming snakes. We find that in swimming snakes, the growth of the amplitude of the propulsive wave head-to-tail is strongly correlated (p < 0.005) with the head-to-tail growth in the wavelength. We investigate correlations between kinematic parameters and steady swimming speed, and find a very strong positive correlation between swimming speed and undulation frequency. We furthermore find a statistically well-supported positive correlation between swimming speed and both the initial amplitude of the propulsive wave at the head and the degree of amplitude growth from head to tail.     

The male red-sided garter snake (Thamnophis sirtalis parietalis): reproductive pattern and behavior

Among the small group of species (e.g., some temperate zone turtles, snakes, and bats) that exhibit a dissociated reproductive pattern, the red-sided garter snake (Thamnophis sirtalis parietalis) is probably the most well studied. For these species, courtship and mating occur immediately upon emergence from winter dormancy; the gonads remain essentially inactive. Male red-sided garter snakes are a particularly informative animal model for examining the role of neuroendocrine factors associated with reproductive physiology and behavior because unlike species that exhibit an associated reproductive pattern, in which sex steroids initiate and control sexual behavior, reproductive behavior in the male garter snake appears to be independent of circulating sex hormone control. In fact, the only factor associated with the initiation of courtship behavior and mating in the male garter snake is an extended period of low temperature dormancy followed by exposure to warm temperatures. Yet the presence of sex steroid-concentrating neurons within the pathways regulating courtship and mating suggests that sex hormones may be involved in the activation of sexual behavior. Although circulating androgens are elevated upon emergence from hibernation, the initiation of courtship behavior and mating appears to be independent of direct androgen control. Thus steroid hormones may have indirect effects on mating behavior in animals that display "dissociated" reproductive behaviors.     

Kinematics of swimming garter snakes (Thamnophis sirtalis).

We investigate the kinematics of swimming garter snakes (Thamnophis sirtalis) using a novel nonlinear regression-based digitization method to establish quantitative statistical support for non-constant wavelengths in the undulatory pattern exhibited by swimming snakes. We find that in swimming snakes, the growth of the amplitude of the propulsive wave head-to-tail is strongly correlated (p < 0.005) with the head-to-tail growth in the wavelength. We investigate correlations between kinematic parameters and steady swimming speed, and find a very strong positive correlation between swimming speed and undulation frequency. We furthermore find a statistically well-supported positive correlation between swimming speed and both the initial amplitude of the propulsive wave at the head and the degree of amplitude growth from head to tail.     

Conflicts between courtship and thermoregulation: the thermal ecology of amorous male garter snakes (Thamnophis sirtalis parietalis, colubridae)

Thermoregulatory behavior is an important component of daily activities for many reptiles, especially for small heliothermic (sun-basking) species that inhabit cold climates. However, the relative costs and benefits of thermoregulation depend on numerous factors, such that reptiles may sometimes accord a low priority to precise control of body temperatures. We observed and radio tracked garter snakes (Thamnophis sirtalis parietalis) in central Manitoba during the mating season (spring). Previous studies on this species have documented precise behavioral regulation of body temperatures during summer. In contrast, the courting snakes that we studied in springtime spent little time in overt thermoregulatory behavior. Body temperatures were extremely variable (both in outdoor enclosures and in the field) despite abundant opportunities for more precise thermal control. These small elongate reptiles cool so quickly (relative to the time periods needed for effective courtship) that any benefit to higher body temperatures would be transitory at best. Experiments show that hotter males are no better at obtaining matings or at detecting predators. Thus, male garter snakes concentrate on courtship rather than on basking. In the face of conflicting priorities, reptiles may often forgo precise thermoregulation because its benefits are too low, and its costs too high, compared with alternative behaviors.     

Mating increases plasma levels of prostaglandin F2 alpha in female garter snakes

Plasma levels of prostaglandin F2 alpha (PGF2 alpha) in female red-sided garter snakes (Thamnophis sirtalis parietalis) were measured at intervals after mating or exposure to males. PGF2 alpha levels increased significantly within 15 minutes of mating and peaked 6-24 hr after mating. Females that did not mate, but received similar amounts of male courtship, had levels of PGF2 alpha significantly lower than those of females that mated. These results extend previous findings that unmated female garter snakes injected with PGF2 alpha exhibit sexual behavior characteristic of females that have mated. Together these data indicate that female garter snakes elaborate PGF2 alpha in response to stimuli associated with mating and that PGF2 alpha has a functional role in inducing post-mating declines in sexual behavior of this species.     

Analysis of variance of first-year growth in captive garter snakes (Thamnophis elegans) by family and sex

Although body size in reptiles has important consequences for their ecology, it is quite variable in most species, presumably because growth in these animals is influenced by many factors. In this study, we analyse the effects of family membership and sex on first-year growth of garter snakes (Thamnophis elegans) held under identical conditions in the laboratory. Litters, but not the two sexes, differed significantly in mean size at birth. Larger snakes ate more food and had a higher absolute (but not relative) growth rate in length than smaller snakes. Thus, there was significant variation in length increase among litters, but not between the sexes. All else being equal, variance in size at birth could contribute significantly to variance in adult body size.     

A proposal to sequence the genome of a garter snake (Thamnophis sirtalis)

Here we develop an argument in support of sequencing a garter snake (Thamnophis sirtalis) genome, and outline a plan to accomplish this. This snake is a common, widespread, nonvenomous North American species that has served as a model for diverse studies in evolutionary biology, physiology, genomics, behavior and coevolution. The anole lizard is currently the only genome sequence available for a non-avian reptile. Thus, the garter snake at this time would be the first available snake genome sequence and as such would provide much needed comparative representation of non-avian reptilian genomes, and would also allow critical new insights for vertebrate comparative genomic studies. We outline the major areas of discovery that the availability of the garter snake genome would enable, and describe a plan for whole-genome sequencing.     

Developmental changes in cytochrome oxidase histochemistry in the main and accessory olfactory bulbs of embryonic and neonatal garter snakes (Thamnophis sirtalis spp.)

Developmental studies examining the changes in oxidative metabolic activity are useful for understanding how and if the vomeronasal and olfactory systems respond to stimulation during embryogenesis. Garter snakes are good candidates for examining the potential functionality of the vomeronasal system in utero. In adult garter snakes, the vomeronasal system mediates many behaviors. Neonatal garter snakes exhibit these same behaviors, and the vomeronasal system has been shown to mediate feeding behavior in neonates. Using cytochrome oxidase histochemistry, we examined changes in the oxidative metabolic activity of main and accessory olfactory bulbs of embryonic and neonatal garter snakes (Thamnophis sirtalis sirtalis and T. s. parietalis). Cytochrome oxidase staining is greater in the accessory olfactory bulb than in the main olfactory bulb of embryonic garter snakes. However, neonates show no differences in the staining of the accessory and main olfactory bulbs, suggesting a change in the stimulation of the main olfactory bulb after birth. This is the first report of cytochrome oxidase histochemistry in reptiles and in the vomeronasal system of embryonic vertebrates. © 1993 Wiley-Liss, Inc.     

An empirical test of evolutionary theories for reproductive senescence and reproductive effort in the garter snake Thamnophis elegans

Evolutionary theory predicts that differential reproductive effort and rate of reproductive senescence will evolve under different rates of external mortality. We examine the evolutionary divergence of age-specific reproduction in two life-history ecotypes of the western terrestrial garter snake, Thamnophis elegans. We test for the signature of reproductive senescence (decreasing fecundity with age) and increasing reproductive effort with age (increasing reproductive productivity per gram female) in replicate populations of two life-history ecotypes: snakes that grow fast, mature young and have shorter lifespans, and snakes that grow slow, mature late and have long lives. The difference between life-history ecotypes is due to genetic divergence in growth rate. We find (i) reproductive success (live litter mass) increases with age in both ecotypes, but does so more rapidly in the fast-growth ecotype, (ii) reproductive failure increases with age in both ecotypes, but the proportion of reproductive failure to total reproductive output remains invariant, and (iii) reproductive effort remains constant in fast-growth individuals with age, but declines in slow-growth individuals. This illustration of increasing fecundity with age, even at the latest ages, deviates from standard expectations for reproductive senescence, as does the lack of increases in reproductive effort. We discuss our findings in light of recent theories regarding the phenomenon of increased reproduction throughout life in organisms with indeterminate growth and its potential to offset theoretical expectations for the ubiquity of senescence.