Functional anatomy of pulmonary ventilation in the garter snake, Thamnophis elegans

The functional anatomy of pulmonary ventilation in Thamnophis elegans was studied by electromyography. Flow of air into and out of the lungs follows a triphasic pattern and occurs while the glottis is held open by a dilator muscle. Non-ventilatory rest periods characterized by a closed glottis and lack of ventilatory movements occur between breaths. Exhalation is caused by contraction of a pair of dorsolateral sheets of muscle that extends from the ventral surface of the vertebral column to the medial surfaces of the ribs and a pair of ventro-lateral sheets that extends from the medial surfaces of the ribs to the mid-ventral skin. Inspiration is powered by contraction of a series of rib levators and retractors. The last phase of the ventilatory cycle is a passive expiration that may be caused by elastic recoil of the walls of the filled lungs and relaxation of muscles used during inspiration. The site of respiratory pumping is shifted craniad after large prey items have been completely ingested. The significance of this shift and its anatomical basis are considered.     

Placentation in the garter snake, Thamnophis sirtalis

The structure and polysaccharide constitution of the jelly capsule of the egg of Rana pipiens is described. Microscopic examination of the jelly capsule revealed the presence of five discrete jelly layers that differed clearly in their response to selected cytochemical tests. These layers were classified as M1-through M5 from the inner to the outermost layer. A sixth layer occasionally could be observed between M3 and M4. All layers contain neutral mucopolysaccharides. In addition layers M1 and M3 contain sulphated mucopolysaccharides, M2 and M4 contain non-sulphated acid mucopolysaccharides, and layer M5 contains both sulphated and non-sulphated acid mucopolysaccharides. M2 may also contain a small quantity of sulphated mucopolysaccharides. The layer that occasionally appears between M3 and M4 is probably an area in which free acidic groups are in higher concentration than in adjacent areas rather than being a discrete jelly layer. Neither hyaluronic acid nor sialic acid was localized by the methods employed. The possible significance of some of these constituents is discussed.     

Pituitary gland of the garter snake (Thamnophis): Morphology and immunocytochemistry

The three-dimensional gross morphology of the pituitary gland of the garter snake (Thamnophis sirtalis) is presented. Hormone-producing cells of the pars distalis were localized immunocytochemically. Corticotropes and lactotropes occur in the anterior two-thirds of the gland; corticotropes are especially numerous in the area of the pars distalis nearest the median eminence, and lactotropes are most abundant medially. Somatotropes are restricted to the posterior one-third of the pars distalis. Gonadotropes and thyrotropes are scattered throughout the pars distalis and in favorable sections form a network of cells enclosing clusters of peptide-secreting cells.     

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.     

The effects of substrate and vertebral number on locomotion in the garter snake Thamnophis elegans

1. Locomotor performance of limbless vertebrates depends on the substrate through which individuals move and may result in selection on vertebral number in different habitats. To evaluate the effect of push-point density on snake locomotion, the density of vegetation and other potential push-points was quantified at two sites in California (coastal and inland), where conspecific snakes differed greatly in vertebral number (230 and 256 average total vertebrae, respectively; Arnold 1988). The coastal site had significantly higher push-point densities than the inland site.
2. Five experimental push-point densities that fell within the natural range of push-point densities were employed in laboratory trials of juvenile snake locomotion. Density of push-points significantly affected both crawling speed and head-to-tail distance (HTD), an indirect measure of lateral bending. The fastest speed was achieved at an intermediate push-point density. The shortest HTD occurred when snakes moved through the lowest push-point density.
3. Sex, total number of vertebrae and total length significantly affected HTD, regardless of push-point density. Snakes with relatively more vertebrae had a shorter HTD, suggesting they were able to achieve greater lateral bending than snakes with fewer vertebrae. Coastal and inland populations did not differ in HTD during locomotion.
4. Numbers of body and tail vertebrae significantly influenced speed at different push-point densities. In general, snakes with more body vertebrae were slower than those with fewer, while snakes with more tail vertebrae were faster than those with fewer. Snakes of greater total length were faster at all densities. Coastal snakes crawled faster than inland snakes at all push-point densities.     

Pulmonary carbonic anhydrase in the garter snake, Thamnophis sirtalis

This study examined whether the snake lung possesses intravascular carbonic anhydrase (CA). Lungs were perfused with control salines and with salines containing CA inhibitors. Perfusion with control salines resulted in a stable CO(2) excretion, whereas CA inhibitors significantly reduced pulmonary CO(2) excretion. Membrane-permeable and membrane-impermeable CA inhibitors produced comparable decreases in CO(2) excretion, suggesting that extracellular, intravascular CA participated in the pulmonary CO(2)-HCO3(-)-H(+) reactions. Treatment of lungs with phosphatidylinositol specific-phospholipase C (PI-PLC) significantly decreased CO(2) excretion, indicating that CA was connected to the luminal endothelial cell membrane by a phosphatidylinositol glycan linkage. Taken together, these results are the first to demonstrate the presence of membrane-bound, intravascular CA (CA IV) in the snake lung.     

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.     

A test of life-history theories of immune defence in two ecotypes of the garter snake, Thamnophis elegans

1.  Life-history theorists have long observed that fast growth and high reproduction tend to be associated with short life span, suggesting that greater investment in such traits may trade off with self-maintenance. The immune system plays an integral role in self-maintenance and has been proposed as a mediator of life-history trade-offs.
2.  Ecoimmunologists have predicted that fast-living organisms should rely more heavily on constitutive innate immunity than slow-living organisms, as constitutive innate defences are thought to be relatively inexpensive to develop and can provide a rapid, general response to pathogens.
3.  We present the first study to examine this hypothesis in an ectothermic vertebrate, by testing for differences in three aspects of constitutive innate immunity in replicate populations of two life-history ecotypes of the garter snake Thamnophis elegans , one fast-living and one slow-living.
4.  As predicted, free-ranging snakes from the fast-living ecotype had higher levels of all three measures of constitutive innate immunity than the slow-living ecotype. These differences in immunity were not explained by parasite loads measured. Furthermore, both ecotypes exhibited a positive relationship between innate immunity and body size/age, which we discuss in the context of ectotherm physiology and ecotype differences in developmental rates.     

Behavioural variation in natural populations. V. Morphological correlates of locomotion in the garter snake (Thamnophis radix)

A series of morphologieal and locomotor performance variables was measured in a population of newborn garter snakes to determine whether performance capacity has a significant morphological basis in these animals. Morphological traits measured were body length and mass, number of body and tail vertebrae and numbers of vertebral abnormalities. Locomotor performances included burst and mid-distance speed and distance and time crawled before anti-predator displays were assumed. All performance variables were repeatable in daily replicate trials ( P < 0.001). Individual burst speed, mid-distance speed, and distance crawled were significantly correlated pairwise ( P < 0.01). Most morphological and performance variables had a significant mass dependence (static allometry), although the effects were rather weak ( r ² < 0.1, except for body length): larger animals performed better and had fewer abnormalities. There were significant associations between some morphological traits and locomotor performance. Morphological factors accounted for 19% of the variation in mid-distance speed and 14% of the variation in antipredator behavior by multiple regression analysis. Canonical correlation of all performance and morphological variables simultaneously accounted for 24% of the observed variation in performance. Numbers of body and tail vertebrae (assayed by scale counts) had an interactive effect on speed of locomotion.     

The photoreceptors and visual pigments of the garter snake (Thamnophis sirtalis): a microspectrophotometric, scanning electron microscopic and immunocytochemical study

Scanning electron microscopy, immunocytochemistry, and single cell microspectrophotometry were employed to characterize the photoreceptors and visual pigments in the retina of the garter snake, Thamnophis sirtalis. The photoreceptor population was found to be comprised entirely of cones, of which four distinct types were identified. About 45.5% of the photoreceptors are double cones consisting of a large principal member joined near the outer segment with a much smaller accessory member. About 40% of the photoreceptors are large single cones, and about 14.5% are small single cones forming two subtypes. The outer segments of the large single cones and both the principal and accessory members of the doubles contain the same visual pigment, one with peak absorbance near 554 nm. The small single cones contain either a visual pigment with peak absorbance near 482 nm or one with peak absorbance near 360 nm. Two classes of small single cones could be distinguished also by immunocytochemistry and scanning electron microscopy. The small single cones with the 360-nm pigment provide the garter snake with selective sensitivity to light in the near ultraviolet region of the spectrum. This ultraviolet sensitivity might be important in localization of pheromone trails. Accepted: 10 March 1997     

Developmental plasticity of immune defence in two life-history ecotypes of the garter snake, Thamnophis elegans - a common-environment experiment

1.?Ecoimmunological theory predicts a link between life-history and immune-defence strategies such that fast-living organisms should rely more on constitutive innate defences compared to slow-living organisms. An untested assumption of this hypothesis is that the variation in immune defence associated with variation in life history has a genetic basis. 2.?Replicate populations of two life-history ecotypes of the garter snake Thamnophis elegans provide an ideal system in which to test this assumption. Free-ranging snakes of the fast-living ecotype, which reside in lakeshore habitats, show higher levels of three measures of constitutive innate immunity than those of the slow-living ecotype, which inhabit meadows around the lake. Although this pattern is consistent with the ecoimmunological pace-of-life hypothesis, environmental differences between the lakeshore and meadow habitats could also explain the observed differences in immune defence. 3.?We performed a common-environment experiment to distinguish between these alternatives. Snakes born and raised in common-environment conditions reflected the immune phenotype of their native habitats when sampled at 4 months of age (i.e. fast-living lakeshore snakes showed higher levels of natural antibodies, complement activity and bactericidal competence than slow-living meadow snakes), but no longer showed differences when 19 months old. 4.?This suggests that the differences in innate immunity observed between the two ecotypes have an important - and likely age-specific - environmental influence, with these immune components showing developmental plasticity. A genetic effect in early life may also be present, but further research is needed to confirm this possibility and therefore provide a more definitive test of the ecoimmunological pace-of-life hypothesis in this system.     

Adaptive divergence within and between ecotypes of the terrestrial garter snake, Thamnophis elegans, assessed with F(ST)-Q(ST) comparisons

Populations of the terrestrial garter snake (Thamnophis elegans) around Eagle Lake in California exhibit dramatic ecotypic differentiation in life history, colouration and morphology across distances as small as a few kilometres. We assayed the role of selection in ecotypic differentiation in T. elegans using F(ST)-Q(ST) analysis and identified selective agents using direct and indirect observations. We extended the conventional implementation of the F(ST)-Q(ST) approach by using three-level analyses of genetic and phenotypic variance to assess the role of selection in differentiating populations both within and between ecotypes. These results suggest that selection has driven differentiation between as well as within ecotypes, and in the presence of moderate to high gene flow. Our findings are discussed in the context of previous correlational selection analyses which revealed stabilizing and correlational selection for some of the traits examined.     

Thermal biology of the common garter snake Thamnophis sirtalis (L.)

Summary The thermoregulatory significance of a striped-melanic colour polymorphism in the common garter snake, Thamnophis sirtalis, was assessed through a combination of labortory experimentation and field study. In experiments with living snakes the melanic morph maintained a higher body temperature than the striped morph, when exposed to natural insolation. Experiments with excised skin showed that this thermal advantage is attributable to some integumental difference between the two morphs. Body temperatures of snakes in the field revealed that, during the colder part of the active season, melanics were able to stay warmer than striped snakes by an amount (1.24 C°) approximating the difference observed in the laboratory. Some evidence and argument is presented to suggest that melanism also may confer protection against overheating in warm periods.     

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.     

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.     

A light microscopic study of the mononuclear cells infiltrating skin homografts in the garter snake, Thamnophis sirtalis (Reptilia: Colubridae)

The rejection of skin homografts in the snake, Thamnophis sirtalis is preceded by an infiltration of mononuclear cells into the graft bed. The initial arrangement of infiltrating cells in perivascular halos suggests that these cells emigrate from the blood stream of the host. A cytological study showed that the vast majority of the cells can be classified as small and mediumsized lymphocytes, monocytes and macrophages. Early stages of infiltration were associated with large proportions of lymphocytes while later stages were characterized by a predominance of macrophages. It was concluded that the mononuclear cells associated with graft rejection include large proportions of lymphocytes and macrophages and not just one kind of lymphoid cell.