Anole Annals

Brown anole with and without crest shown at the whole-animal (left) and histological (right) level. Photo from Ademi and Rand poster.

If you’ve ever been around brown anoles, you know that the males can be pretty aggressive. Part of that aggression involves the enlargement of a crest along the neck and back. The crest is caused by fluid rapidly rushing into the tissue of the crest. How this works has been discussed here before, but Matt Rand’s research group at Carleton College continues to try to unravel what hormonal pathways are responsible for crest formation. Ademi and Rand used an experimental approach to discover what molecular receptors are activated to cause crest formation. Body-wide and local injection of a variety of chemicals and drugs gave some tantalizing clues as to how it works.

Local injections to stimulate cAMP activity caused crest formation locally (top), whereas body-wide injection caused whole-crest formation (bottom). Photo from Ademi and Rand poster.

They found after several inhibitory and stimulatory drug manipulations that crest erection is likely stimulated by epinephrine acting on a Beta-2 like adrenergic receptor that stimulates cyclic AMP (cAMP) activity to cause vasodilation (enlarging of blood vessels) and fluid entrance into the crest. This activity that starts with the B2-adrenergic receptor is essentially the same function as that seen in mammalian circulatory systems, including us. They also stimulated cAMP activity without stimulating the B2-like adrenergic receptor and found similar results. You can see how dramatic the response was below, where they used local injection to cause crest formation only at the site of injection! The use of epinephrine binding to a B2-like adrenergic receptor as the molecule of communication makes the rapid time-course of crest formation make sense. There are still some unknown aspects as to how the vasodilation mechanistically causes the fluid release in the crest, but they are actively studying it.

Are anoles like sherpas? Photo from Reddit

When you’re used to living at low to moderate elevations, it can be challenging to visit high-altitude places. The declining partial pressure of oxygen at high altitude makes it difficult for your body to deliver the same amount of oxygen to tissues. This is why National Football League players often struggle to play in Denver (see playoffs next week!). However, organisms that live at high elevations, including humans, have evolved a number of ways to deal with living in such oxygen-challenged environments. We know less about whether the same aspects of the cardiovascular change in different organisms, even among relatively closely related species. Well, what better group of organisms to address such questions than anoles!

Virtually nobody reading this blog will be unfamiliar with the story of the Greater Antillean ecomorphs, and they are great to use for questions related to elevation and adaptations to deal with it. They live along steep elevational gradients within an island, and such gradients exist across islands. Although, the Caribbean anoles have been the subject of numerous studies that have shown convergent evolution in body size and shape, as well as locomotor performance and endocrine function, we know much less about how they deal with elevational challenges at the cardiovascular level.

Species studied and locations in the Dominican Republic. Photo from Webber et al.’s poster.

Miguel Webber, an undergraduate in the laboratory of Michele Johnson at Trinity University, along with Brittney Ivanov, studied several blood physiology traits in 13 species across five ecomorphs in the Dominican Republic to determine whether elevation has been an important driving force in the evolution of oxygen delivery mechanisms. Although looking at an impressive number of traits that included hematocrit (the proportion of red blood cells), hemoglobin concentration, and red blood cell size, Miguel only found hemoglobin concentration to be positively related with elevation when looking across species.

One of the more interesting findings was that none of the blood physiology variables that Miguel measured were ecomorph specific. However, this makes sense because members of an ecomorph live across wide geographic areas and across elevational gradients. Physiological studies such as Miguel’s are offering interesting insights into how anoles have adapted to their environments and emphasizes that ecomorph membership does not determine everything.

Shane Campbell-Staton gives his talk at SICB 2016.

Climate change isn’t just leading to greater average environmental temperatures – it’s also leading to an increase in the frequency and severity of extreme weather events, such as heat waves and hurricanes. Of interest to Shane Campbell-Staton, a post-doctoral researcher in the Cheviron Lab and a recent graduate from the Losos Lab at Harvard, is understanding how the recent polar vortex in North America impacted the native green anole, Anolis carolinensis. The polar vortex of winter 2013/2014 set several records in snow fall and in all-time low temperatures in the south, and also led to severe weather in the midwest and east.

Shane found that, immediately following the polar vortex event, cold tolerance (CTmin) was significantly lower in lizards from southern Texas, as low as in lizards from much higher latitudes. He suggested that this result stems from differential survivorship during the event – lizards in south Texas that were more cold tolerant (i.e., had a lower CTmin) were more likely to survive the winter vortex  than less cold tolerant individuals. He then returned to south Texas a few months later and sampled both the survivors and their offspring and found that the decrease in CTmin persisted, indicating a potential evolutionary shift in cold tolerance. He put the final nail in the coffin by running a common garden experiment, where he demonstrated that, even when reared under common laboratory conditions, offspring exhibited cold tolerance similar to their parents, indicating high heritability in this trait and that the shift observed in nature was evolved rather than due to plasticity.

Shane then examined the response to the weather event at the genetic level by sequencing liver transcriptomes. Transcriptomes quantify patterns of gene expression levels for all genes regulated in a tissue; hence, by examining what genes are differentially expressed following cold stress, we can figure out the molecular underpinnings to cold adaptation and acclimation. He found that gene expression in survivors from the south closely resembled expression patterns in northern lizards, indicating a shared molecular pathway to cold tolerance adaptation in lizards from both habitats. The gene expression modules (or groups of genes) that exhibited a strong statistical association with CTmin variation were overrepresented for genes associated with oxidative phosphorylation. Oxygen consumption, which feeds oxidative phosphorylation, is directly related to CTmin: Animals that are more cold tolerant consume less oxygen during cooling. Hence, the expression differences in oxidative phosphorylation may pinpoint a proximate mechanism for cold tolerance adaptation.

You can learn more about Shane’s work on adaptation following the polar vortex in his recent Harvard Horizons talk.

*The following post was written by Chris Robinson, a Master’s student in Matt Gifford’s lab at the University of Central Arkansas.*

Like at every SICB conference, anoles are well represented among the talks and posters here in Portland and we here at the Anole Annals couldn’t be more thrilled to see the love for one of our favorite genera.

Christopher Anderson, a post-doctoral associate in Thomas Roberts’ lab at Brown University, gave a talk examining how muscle physiology influences whole organism performance in five species of anoles. His group examined two muscles, the M. ambiens pars ventralis (a swing phase muscle of the leg important for locomotion) and the M. abductor mandibulae externus superficialis anterior (a muscle in the jaw used in biting), to see if they differed from each other in how they perform in order to meet their functional demands. The muscle of the leg, which is used in sprinting, is cyclically activated and deactivated as an organism moves, whereas the muscle of the jaw is used more episodically.

Anderson and his colleagues found that the leg muscle builds passive tension at shorter lengths and has a twitch time that is 1.3-2.0 times faster than that of the jaw (to see how twitch time relates to sprint speed, see the post about Noel Parks’ poster). From this, Anderson concluded that these muscles are tuned to meet their physiological demands. Locomotion muscles, which are used frequently, generate a lot of force rapidly and the quickly developing passive tension in these muscles may serve as a form of protection for the muscle during active lengthening.

Hanna Wegener giving her talk on Anolis sagrei from the Bahamas at SICB 2016

Small islands are great systems in which to study evolution, in part because their isolation and simplified landscapes makes them amenable to experimental studies. For example, previous experimental work on Anolis sagrei in the Bahamas by Losos et al. (1997) and Kolbe et al. (2012) found evolutionary changes in hindlimb length driven by adaptation to structural habitat over only a few years.

Hanna Wegener, a Ph.D. student studying with Jason Kolbe at the University of Rhode Island, wanted to know if morphological differences associated with habitat use also manifest in natural (rather than experimentally introduced) populations of Anolis sagrei from the Bahamas. She examined genetic (microsatellite) and morphological variation from male and female A. sagrei on seventeen islands in the Bahamas. Despite the islands being separated by very small geographic distances (no more than three kilometers and typically only a few hundred meters), populations on the islands were genetically differentiated. Her genetic analysis further found high levels of inbreeding on each island.

Unlike the findings on the experimental islands, Hanna did not find any correlation between perch diameter and hindlimb length. She did find that female density was high on the islands, and that density correlated strongly with head length and injury frequency, suggesting that competition influences morphological differentiation on these islands. Overall, Hanna found that morphological patterns varied considerably among islands and among males and females. She suggests that this variation is due to stochastic effects on small islands, namely genetic drift, due to the extinction and colonization dynamics in response to hurricanes.

SICB is off to a very anole-y start in Portland! There have been anole-focused talks and posters all day, and your intrepid team of AA reporters are on the scene.

At Monday’s poster session, Noel Parks (an undergraduate at Brown University working with Chris Anderson and Thomas Roberts) presented her research on muscle contraction and sprint kinematics in Anolis sagrei and A. cristatellus. The team performed laboratory sprint trials with the two species at a range of inclines, and then using muscle tissues from the same lizards used in the trials, they measured how fast the M. ambiens pars ventralis (a hindlimb muscle critical for locomotion) can contract and relax after stimulation, a measure they call muscle twitch time.

Noel Parks and her poster at SICB 2016.

For both species, Noel and her colleagues found that stance time (the amount of time a foot is in contact with the ground) and swing time (the amount of time the limb is moving forward) are limited by the muscle twitch time. Thus, muscle twitch time may constrain the sprint speed of these animals. Further, at steeper inclines, stance and swing times more closely approached muscle twitch time. The two species differed in these speeds, however, as A. sagrei had faster twitch, stance, and swing times than A. cristatellus.

This work gives us another interesting piece of the puzzle in the larger story of anole locomotor performance!