Category: New Research Page 31 of 67

Why the long face?

When most people think of vertebrate sexual dimorphism (differences between the sexes), they think of elephant seals or red deer. Most of us here, of course, think of the pronounced dimorphism in size and shape in many anole species. Indeed, anoles have served as excellent model systems for the study of sexual dimorphism, particularly the evolutionary forces that give rise to it. Although there has been significant progress since Darwin in our understanding of why sexual dimorphism evolves, we have made less progress in the HOW. That is, what mechanisms during development give rise to what are often extreme differences between the sexes when their genomes are so similar?

When we think of vertebrates where males are larger or shaped differently than females, and have weapons or ornaments, we almost immediately think of testosterone as a mechanism underlying the sex differences. Once sexual maturity happens, the testes start cranking out testosterone, thus causing a change in the male’s phenotypic trajectory. While there is certainly evidence for circulating testosterone to have this effect in some lizards, is this always the case, and does it apply to specific body parts and not just overall size? Aside from the circulating hormone, how are receptors involved in the development of dimorphism? In a new paper by Sanger et al., a novel developmental pathway of sexual dimorphism is described for lizards in the carolinensis clade, which are striking in their elongation of male faces relative to females.

Figure 1a. from Sanger et al. (2014), showing the differences in head shape dimorphism among anole clades. Note the long male face in A. maynardi, a member of the carolinensis clade.

Sanger et al. tested whether sex differences in several different pathways led to the observed head shape dimorphism in A. carolinensis compared to two non-carolinensis species (A. cristatellus and A. sagrei) that exhibit shorter male faces. They show, using a combination of developmental and molecular genetic techniques, that the extreme elongation of male heads in carolinensis lizards is not due to an androgen pathway (i.e., testosterone) or the somatropic axis (i.e., insulin-like growth factor). Instead, they found a significant shift in the estrogen pathway. Specifically, at sexual maturity, males decrease expression of estrogen receptors (erβ), which is the beginning of a signaling cascade, ultimately resulting in up-regulation of genes involved in skeletogenesis in the skull of males.

Figure 4 from Sanger et al. (2014), showing the molecular pathway underlying facial elongation in A. carolinensis.

This identification of a novel mechanism for the development of sexual dimorphism will certainly stimulate further evo-devo research in anoles and beyond. For starters, is the same pathway responsible for male facial elongation in other species in the carolinensis clade, or are more ‘traditional’ mechanisms operating there? This important research highlights that investigators need to consider all aspects of signaling systems, including circulating hormones, their receptors, and signal cascades that result from activation of a particular pathway. Clearly this paper by Sanger et al. is an excellent step in the right direction for understanding how developmental pathways lead to adult difference in anoles, and it will also steer other investigators to consider a diversity of developmental mechanisms in their quest to elucidate how adults end up the way they do.

Sanger TJ, Seav SM, Tokita M, Langerhans RB, Ross LM, Losos JB, Abzhanov A. 2014. The oestrogen pathway underlies the evolution of exaggerated male cranial shapes in Anolis lizards. Proceedings of the Royal Society B 281:20140329.

Mothers affect the quality of their offspring. As humans, this seems obvious. For example, expecting mothers often take prenatal vitamins, limit their consumption of certain foods, and avoid kitty litter knowing that these minor environmental factors can affect the normal development of the fetus. Related statements could be made for the relationship of a mother and child after birth. Understanding the precise effects that parents have on their offspring has been of great interest to biologists from many disciplines as they disentangle the genetic and environmental factors that underlie differential survival and reproduction for individuals within a population (fitness). Because Anolis lizards can be easily maintained in captivity and their eggs readily manipulated they provide a useful model for the examination of maternal effects. Warner and Lovern took advantage of these qualities and tested the role of maternal body condition on offspring quality in the brown anole, Anolis sagrei.

A. sagrei from Cayman Brac

Nutritional stress is a well-studied example of how maternal condition may affect juvenile quality; if the mother is malnourished the quality of her egg yolk may suffer, which, in turn, affects embryonic development. The authors tested this hypothesis in A. sagrei by manipulating the amount of food gravid females received, feeding approximately 168 crickets per lizard in a “high-prey” treatment versus 84 crickets in a “low-prey” treatment distributed over 11 weeks. During this time the authors carefully assessed the number and size (mass) of the eggs and, subsequently, the quality (mass-and-snout to vent length) of the hatchlings. Impressively, the authors didn’t stop there. They also experimentally manipulated the amount of nutrition in a subset of eggs by removing yolk with a syringe. Followed by a battery of statistical models, this study is quite a nice physiological analysis that has evolutionary implications.

When comparing the two diet regimes, Warner and Lovern found that body condition does affect the quality of offspring; females maintained on the “low-prey” diet produced eggs 6.6% smaller than females raised on the “high-prey” diet. In turn, smaller eggs also tended to hatch more quickly and smaller eggs produced smaller hatchlings, both probably due to the lower amount of available nutrition (paradoxically, neither incubation time or hatchling mass was directly correlated with maternal prey availability). Low prey availability also results in hatchlings with slower growth rates. The experimental reduction of egg yolk supports the results of the prey availability study: hatchlings from yolk-reduced females were 8% shorter and 23% lighter and grew more slowly than those hatched from unmanipulated eggs. It is clear from their results that nutrition has an effect on hatchling quality well into life, after the obvious maternal effects have passed. There are a number of other interesting correlations (and statistical caveats) described within the text that may also be of interest to some readers.

Figure 5 from Warner and Lovern 2014.

What is becoming clear from studies like these is that environmental stressors can have lasting effects on organismal development that transcend generational boundaries. Mechanistic studies, such as those on the American alligator, illustrate that these effects are mediated by heritable methylation patterns of key regulatory genes. The stressors do not need to be long lasting; physiological responses can result from acute events that occur within key developmental windows, often when a particular organ is maturing. While stressing the embryo too far results in abnormal embryonic development, more subtle effects may not arise until late in life or subsequent generations. Anoles, and A. sagrei in particular, may provide a number of opportunities for environmental health research in the future. Studies such as the one described above could be performed to more precisely dissect the organ-specific effects of maternal nutritional stress or whether the effects dissipate with age. Similar to the alligator studies, eggs laid in polluted soils may allow opportunities for developmental toxicology research. Growing genomic resources may allow for examination of genome-wide and gene-specific methylation patterns within and outside of polluted habitats. The possibilities are broad and the impact cannot be predicted at this time, but the potential is there for much more detailed mechanistic research on the relationship between developmental physiology and the environment.

When it rains, it pours. Research on the immense diversity in anole chromosomes was rampant in the 1970’s and early 1980’s, and then…nothing. Until, that is, the last two months. Not one, but two, papers appeared in Evolution, and now AA has learned of a paper on chromosomal variation in Norops clade anoles, recently published in Zoological Studies (click for a downloadable pdf). The paper, by Castiglia et al., examines karyotypes in Norops anoles and argues that karyological variation is in some cases consistent with our understanding of phylogenetic relationships within the group.

Abstract
Background: Neotropical lizards, genus Anolis (Polychrotidae), with nearly 380 species, are members of one of the most diversified genera among amniotes. Herein, we present an overview of chromosomal evolution in ‘beta’ Anolis (Norops group) as a baseline for future studies of the karyotypic evolution of anoles. We evaluated all available information concerning karyotypes of Norops, including original data on a recently described species, Anolis unilobatus. We used the phylogeny of Norops based on DNA sequence data to infer the main pattern of chromosomal evolution by means of an ancestral state analysis (ASR).

Results: We identified 11 different karyotypes, of which 9 in the species had so far been used in molecular studies. The ASR indicated that a change in the number of microchromosomes was the first evolutionary step, followed by an increase in chromosome numbers, likely due to centric fissions of macrochromosomes. The ASR also showed that in nine species, heteromorphic sex chromosomes most probably originated from six independent events.

Conclusions: We observed an overall good correspondence of some characteristics of karyotypes and species relationships. Moreover, the clade seems prone to sex chromosome diversification, and the origins of five of these heteromorphic sex chromosome variants seem to be recent as they appear at the tip nodes in the ancestral character reconstruction. Karyotypic diversification in Norops provides an opportunity to test the chromosomal speciation models and is expected to be useful in studying relationships among anole species and in identifying cryptic taxa.

Photo from http://www.frogtown.org/

For more than three decades, since the seminal work of Ray Huey, Al Bennett, and Steve Arnold, biologists have measured whole animal performance–how fast they run, how far they jump, how well they can swim–to understand how species are adapted to their environment.  Work on anoles has been a prime example of how we can study differences among individuals and species to understand how natural selection works and why species living in different environments possess different morphologies (several AA posts have discussed this sort of work [e.g., 1, 2, 3]).

But a critical assumption of all of this research is that we can get animals to perform maximally. Otherwise, it’s tough to study what causes variation in maximal capabilities if animals aren’t performing maximally. The catch is: how do you tell if an animal is going all out? Sure, it’s easy to weed out the slackers, but distinguishing a lizard giving it his all from one going at, say, 90% of max…hard to tell.

In an important and entertaining paper, Henry Astley and colleagues provide some sobering information. The short story goes as follows, and you really should watch the video below for more details and some great images: biomechanicians have studied frog jumping for decades to understand how muscles work. Bullfrogs are known not to jump very well. The maximum jump ever recorded in the lab was only 1.3 m, whereas the much smaller Cuban treefrog can bound 1.7 m. The proffered explanation was that bullfrogs live on land and in the water, and so their morphology must be a compromise.

But…the Guinness Book of World Records claims that a bullfrog–Rosie the Ribeter, to be exact–once jumped 2.18 meters at the Calaveras County Fair. That’s  68% farther than any scientist had ever recorded in the lab. Sounds like a bunch of hooey, right? Well, just to debunk this nonsense, a bunch of Brown University biologists headed to sunny California to visit the County Fair, eat some cotton candy, and check out the frogs. And, lo and behold, it’s true–bullfrogs there regularly far exceed the lab record.

The story’s a lot more complicated–it turns out that there are “pro” frog jumpers–and I won’t go into the details; the paper is well worth a read, very entertaining and sobering for lab performance types (abstract here). But the short story is this: it seems that lab studies had massively underestimated how far bullfrogs can jump, calling into question many of the conclusions that had been reached about their physiology. Moreover, records for the maximum jump distance at the fair showed a steady increase for the first 50 years before levelling off for the last 30. This suggests that the people who jump the frogs (and some families have been doing this for generations) have only gradually learned exactly what conditions and behaviors maximally stimulate the frogs. And this suggests that lab scientists, who just guess at what may work best and tinker a little bit, may not have much of a chance of hitting on the right stimuli.

There’s been lots of great press coverage, too–just google “calaveras frog astley” or something like that. But, first, watch the video and go read the paper (I can email you a copy if you can’t access it online).

httpv://www.youtube.com/watch?v=QKFpvoez7_M