Kevin E. Bonine

Muscle fiber-type variation in lizards (Squamata) and phylogenetic reconstruction of hypothesized ancestral states

SUMMARY Previously, we found that phrynosomatid lizards, a diverse group common in the southwestern USA, vary markedly in fiber-type composition of the iliofibularis (a hindlimb muscle important in locomotion). Phrynosomatidae comprises three subclades: the closely related sand and horned lizards, and their relatives the Sceloporus group. The variation in muscle fiber-type composition for 11 phrynosomatid species is attributable mainly to differences between the sand- and horned-lizard subclades. Here, we expand the phrynosomatid database with three additional species and compare these results with data collected for 10 outgroup (distantly related) species. Our goal was to determine if the patterns found in Phrynosomatidae hold across a broader phylogenetic range of the extant lizards and to elucidate the evolution of muscle fiber-type composition and related traits. To allow for meaningful comparisons, data were collected from species that are primarily terrestrial and relatively small in size (3.5–65 g body mass). Results indicate that the fiber-type variation observed within the Phrynosomatidae almost spans the range of variation observed in our sample of 24 species from eight families. However, one species of Acanthodactylus (Lacertidae) had a consistent region of large tonic fibers (that did not stain darkly for either succinic dehydrogenase or myosin ATPase activity), a fiber-type only occasionally seen in the other 23 species examined. Many species have a large proportion of either fast-twitch glycolytic (FG; e.g. sand lizards and Aspidoscelis) or fast-twitch oxidative-glycolytic (FOG) fibers (e.g. horned lizards), with the slow-oxidative proportion occupying only 1–17% of the iliofibularis. Importantly, the negative relationship between FG and FOG composition observed in Phrynosomatidae appears to be a characteristic of lizards in general, and could lead to functional trade-offs in aspects of locomotor performance, as has previously been reported for Lacertidae. Reconstruction of ancestral trait values by use of phylogenetically based statistical methods indicates especially large changes in fiber-type composition during the evolution of horned lizards.

Tectorial Membrane Morphological Variation: Effects upon Stimulus Frequency Otoacoustic Emissions

The tectorial membrane (TM) is widely believed to play an important role in determining the ears ability to detect and resolve incoming acoustic information. While it is still unclear precisely what that role is, the TM has been hypothesized to help overcome viscous forces and thereby sharpen mechanical tuning of the sensory cells. Lizards present a unique opportunity to further study the role of the TM given the diverse inner-ear morphological differences across species. Furthermore, stimulus-frequency otoacoustic emissions (SFOAEs), sounds emitted by the ear in response to a tone, noninvasively probe the frequency selectivity of the ear. We report estimates of auditory tuning derived from SFOAEs for 12 different species of lizards with widely varying TM morphology. Despite gross anatomical differences across the species examined herein, low-level SFOAEs were readily measurable in all ears tested, even in non-TM species whose basilar papilla contained as few as 50-60 hair cells. Our measurements generally support theoretical predictions: longer delays/sharper tuning features are found in species with a TM relative to those without. However, SFOAEs from at least one non-TM species (Anolis) with long delays suggest there are likely additional micromechanical factors at play that can directly affect tuning. Additionally, in the one species examined with a continuous TM (Aspidoscelis) where cell-to-cell coupling is presumably relatively stronger, delays were intermediate. This observation appears consistent with recent reports that suggest the TM may play a more complex macromechanical role in the mammalian cochlea via longitudinal energy distribution (and thereby affect tuning). Although significant differences exist between reptilian and mammalian auditory biophysics, understanding lizard OAE generation mechanisms yields significant insight into fundamental principles at work in all vertebrate ears.

Tracing Genetic Lineages of Captive Desert Tortoises in Arizona

Abstract We genotyped 180 captive desert tortoises (Gopherus agassizii) from Kingman (n  =  45), Phoenix (n  =  113), and Tucson (n  =  22), Arizona, USA, to determine if the genetic lineage of captives is associated with that of wild tortoises in the local area (Sonoran Desert). We tested all samples for 16 short tandem repeats and sequenced 1,109 base pairs of mitochondrial DNA (mtDNA). To determine genetic origin, we performed assignment tests against a reference database of 997 desert tortoise samples collected throughout the Mojave and Sonoran Deserts. We found that >40% of our Arizona captive samples were genetically of Mojave Desert or hybrid origin, with the percentage of individuals exhibiting the Mojave genotype increasing as the sample locations approached the California, USA, border. In Phoenix, 11.5% were Sonoran–Mojave crosses, and 8.8% were hybrids between desert tortoise and Texas tortoise (G. berlandieri). Our findings present many potential implications for wild tortoises in the Sonoran Desert of Arizona. Escaped or released captive tortoises with Mojave or hybrid genotypes have the potential to affect the genetic composition of Sonoran wild populations. Genotyping captive desert tortoises could be used to inform the adoption process, and thereby provide additional protection to native desert-tortoise populations in Arizona.

Speed and Endurance Do Not Trade Off in Phrynosomatid Lizards

Trade-offs are a common focus of study in evolutionary biology and in studies of locomotor physiology and biomechanics. A previous comparative study of 12 species of European lacertid lizards found a statistically significant negative correlation between residual locomotor speed and stamina (controlling for variation in body size), consistent with ideas about trade-offs in performance based on variation in muscle fiber type composition and other subordinate traits. To begin examining the generality of this finding in other groups of squamates, we measured maximal sprint running speed on a high-speed treadmill and endurance at 1.0 km/h (0.28 m/s) in 14 species of North American phrynosomatid lizards, plus a sample of nine additional species to encompass some of the broadscale diversity of lizards. We used both conventional and phylogenetically informed regression analyses to control for some known causes of performance variation (body size, stockiness, body temperature) and then computed residual performance values. We found no evidence for a trade-off between speed and endurance among the 14 phrynosomatids or among the 23 species in the extended data set. Possible explanations for the apparent difference between lacertids and phrynosomatids are discussed.

Coupled, Active Oscillators and Lizard Otoacoustic Emissions

The present study empirically explores the relationship between spontaneous otoacoustic emissions (SOAEs) and stimulus‐frequency emissions (SFOAEs) in lizards, an ideal group for such research given their relatively simple inner ear (e.g., lack of basilar membrane traveling waves), diverse morphology across species/families (e.g., tectorial membrane structure) and robust emissions. In a nutshell, our results indicate that SFOAEs evoked using low‐level tones are intimately related to underlying SOAE activity, and appear to represent the entrained response of active oscillators closely tuned to the probe frequency. The data described here indicate several essential features that are desirable to capture in theoretical models for auditory transduction in lizards, and potentially represent generic properties at work in many different classes of “active” ears.

Evaluation of Canoe Surveys for Anurans along the Rio Grande in Big Bend National Park, Texas

Abstract Surveys for amphibians along large rivers pose monitoring and sampling problems. We used canoes at night to spotlight and listen for anurans along four stretches of the Rio Grande in Big Bend National Park, Texas, in 1998 and 1999. We explored temporal and spatial variation in amphibian counts and species richness and assessed relationships between amphibian counts and environmental variables, as well as amphibian-habitat associations along the banks of the Rio Grande. We documented seven anuran species, but Rio Grande leopard frogs (Rana berlandieri) accounted for 96% of the visual counts. Chorus surveys along the river detected similar or fewer numbers of species, but orders of magnitude fewer individuals compared to visual surveys. The number of species varied on average by 37% across monthly and nightly surveys. We found similar average coefficients of variation in counts of Rio Grande leopard frogs on monthly and nightly bases (CVs = 42–44%), suggesting that canoe surveys are a fairly precise technique for counts of this species. Numbers of Rio Grande leopard frogs observed were influenced by river gage levels and air and water temperatures, suggesting that surveys should be conducted under certain environmental conditions to maximize counts and maintain consistency. We found significant differences in species richness and bullfrog (Rana catesbeiana) counts among the four river stretches. Four rare anuran species were found along certain stretches but not others, which could represent either sampling error or unmeasured environmental or habitat differences among the river stretches. We found a greater association of Rio Grande leopard frogs with mud banks compared to rock or cliff (canyon) areas and with seepwillow and open areas compared to giant reed and other vegetation types. Canoe surveys appear to be a useful survey technique for anurans along the Rio Grande and may work for other large river systems as well.

Characterization of STR/microsatellite primers for the Gila monster, Heloderma suspectum screened from paired-end Illumina shotgun sequencing

We used Illumina paired-end shotgun sequencing to characterize microsatellite loci in Heloderma suspectum. We identified over 124,000 potentially amplifiable loci and describe PCR primers for 18 variable tri- and tetra-nucleotide STRs. The observed number of alleles per locus ranged from 5 to 16 and heterozygosity varied from 0.64 to 0.92. In addition 13 of these loci cross-amplified in the beaded lizard, Heloderma horridum. This method of microsatellite identification proved extremely efficient and cost effective. This novel marker set can be used by researchers to better understand these elusive species.