M. Rockwell Parker

Social behavior and pheromonal communication in reptiles.

The role of pheromones in orchestrating social behaviors in reptiles is reviewed. Although all reptile orders are examined, the vast majority of the literature has dealt only with squamates, primarily snakes and lizards. The literature is surprisingly large, but most studies have explored relatively few behaviors. The evolution of chemical signaling in reptiles is discussed along with behaviors governed by pheromones including conspecific trailing, male-male agonistic interactions, sex recognition and sex pheromones, and reptilian predator recognition. Nonreptilian prey recognition by chemical cues was not reviewed. The recent literature has focused on two model systems where extensive chemical ecology studies have been conducted: the reproductive ecology of garter snakes and the behavioral ecology of Iberian lacertid lizards. In these two systems, enough is known about the chemical constituents that mediate behaviors to explore the evolution of chemical signaling mechanisms that affect life history patterns. In addition, these models illuminate natural and sexual selection processes which have lead to complex chemical signals whose different components and concentrations provide essential information about individuals to conspecifics. Reptiles provide excellent candidates for further studies in this regard not only in squamates, but also in the orders where little experimental work has been conducted to date.

Genetic loss or pharmacological blockade of testes-expressed taste genes causes male sterility

TAS1R taste receptors and their associated heterotrimeric G protein gustducin are involved in sugar and amino acid sensing in taste cells and in the gastrointestinal tract. They are also strongly expressed in testis and sperm, but their functions in these tissues were previously unknown. Using mouse models, we show that the genetic absence of both TAS1R3, a component of sweet and amino acid taste receptors, and the gustducin α-subunit GNAT3 leads to male-specific sterility. To gain further insight into this effect, we generated a mouse model that expressed a humanized form of TAS1R3 susceptible to inhibition by the antilipid medication clofibrate. Sperm formation in animals without functional TAS1R3 and GNAT3 is compromised, with malformed and immotile sperm. Furthermore, clofibrate inhibition of humanized TAS1R3 in the genetic background of Tas1r3−/−, Gnat3−/− doubly null mice led to inducible male sterility. These results indicate a crucial role for these extraoral “taste” molecules in sperm development and maturation. We previously reported that blocking of human TAS1R3, but not mouse TAS1R3, can be achieved by common medications or chemicals in the environment. We hypothesize that even low levels of these compounds can lower sperm count and negatively affect human male fertility, which common mouse toxicology assays would not reveal. Conversely, we speculate that TAS1R3 and GNAT3 activators may help infertile men, particularly those that are affected by some of the mentioned inhibitors and/or are diagnosed with idiopathic infertility involving signaling pathway of these receptors.

A novel mechanism regulating a sexual signal: The testosterone-based inhibition of female sex pheromone expression in garter snakes

Vertebrates communicate their sex to conspecifics through the use of sexually dimorphic signals, such as ornaments, behaviors and scents. Furthermore, the physiological connection between hormones and secondary sexual signal expression is key to understanding their dimorphism, seasonality and evolution. The red-sided garter snake (Thamnophis sirtalis parietalis) is the only reptile for which a described pheromone currently exists, and because garter snakes rely completely on the sexual attractiveness pheromone for species identification and mate choice, they constitute a unique model species for exploring the relationship between pheromones and the endocrine system. We recently demonstrated that estrogen can activate female pheromone production in male garter snakes. The purpose of this study was to determine the mechanism(s) acting to prevent female pheromone production in males. We found that castrated males (GX) are courted by wild males in the field and produce appreciable amounts of female sex pheromone. Furthermore, pheromone production is inhibited in castrates given testosterone implants (GX+T), suggesting that pheromone production is actively inhibited by the presence of testosterone. Lastly, testosterone supplementation alone (T) increased the production of several saturated methyl ketones in the pheromone but not the unsaturated ketones; this may indicate that saturated ketones are testosterone-activated components of the garter snakes skin lipid milieu. Collectively, our research has shown that pheromone expression in snakes results from two processes: activation by the feminizing steroid estradiol and inhibition by testosterone. We suggest that basal birds and garter snakes share common pathways of activation that modulate crucial intraspecific signals that originate from skin.

Ethnic/Racial and Genetic Influences on Cerumen Odorant Profiles

This report describes the volatile organic compounds (VOCs) associated with human cerumen (earwax) and the effects of ethnicity/race and variation on the ATP-binding cassette, sub-family C, member 11 gene (ABCC11). A single nucleotide polymorphism (SNP) in ABCC11 affects the cerumen VOC profiles of individuals from African, Caucasian, and Asian descent. Employing gas chromatography/mass spectrometry (GC/MS) we have identified the nature and relative abundance of cerumen VOCs from 32 male donors. Our results show that cerumen contains a complex mixture of VOCs and that the amounts of these compounds vary across individuals as well as across ethnic/racial groups. In six of the seven compounds whose detected concentrations were found to be statistically different across groups, individuals of African descent (AfD) > Caucasian descent (CaD) > Asians descent (AsD). Our findings also reveal that ABCC11 genotype alone does not predict the type and relative levels of volatiles found in human cerumen, and suggest that other biochemical pathways must be involved. Examination of the composition and diversity of external auditory canal microbiota in a small subset of our subject population revealed that the ear microbiota may not be directly correlated with either ethnic group membership or ABCC11 genotype.

Itpr3 Is Responsible for the Mouse Tufted (tf) Locus

The tf (tufted) locus is responsible for a classic phenotype of hair loss and regrowth in mice. It is a characteristic of the BTBR strain. Here, we use a combination of positional cloning methods and complementation mapping to identify Itpr3, the inositol triphosphate receptor type 3, as the gene responsible for the tf locus.

THE ROLE OF AIRBORNE AND SUBSTRATE CUES FROM NON-ENVENOMATED MICE DURING RATTLESNAKE (CROTALUS OREGANUS) POST-STRIKE TRAILING

Rattlesnakes are highly adept at following substrate chemosensory trails left by struck (envenomated) prey. However, there are other sources of chemical information that may be present in the post-strike environment, such as odors from unstruck mice, that could either confound or aid predatory success. Rattlesnakes can follow airborne chemosensory trails of a struck prey, but the ability of rattlesnakes to use airborne chemosensory cues of unstruck prey has not been examined. Therefore, our purpose was to examine the abilities of rattlesnakes to locate prey using substrate and airborne cues from unstruck mice and compare this with their success with airborne odors from struck mice. We used a Plexiglas Y-maze through which paired airborne and/or substrate odor choices were presented to northern Pacific rattlesnakes (Crotalus oreganus). In one treatment, snakes given a choice of odor from an unstruck mouse (substrate) versus a control (unscented air) showed a significant preference for the unstruck, substrate odor trail. In a second treatment, snakes given a choice of unstruck mouse odor (airborne) versus a control (unscented air) showed no significant preference for the airborne odor. In a third treatment, snakes given a choice of unstruck mouse (airborne) versus struck mouse odor (airborne) again showed no preference for either odor. This last result suggests that one cue used during airborne trailing is a general chemical cue and is most likely not associated with a distinct post-strike searching image (as is the case in substrate trailing). Snakes also exhibited certain behaviors within the maze (i.e., turning behavior, emergences) more often when exposed to airborne cues than when exposed to substrate cues. These observations indicate that a different system of behaviors is used when rattlesnakes follow airborne compared to substrate odor cues during post-strike trailing.

Behavioral variation in prey odor responses in northern pine snake neonates and adults

Squamate reptiles (snakes, lizards, amphisbaenians) rely heavily on chemosensory cues to identify, locate and choose between suitable prey items, but comparatively little research has focused on the chemical ecology of threatened squamate species. Such knowledge highlights ecologically important aspects of their survival. Due to gape limitations, squamates often demonstrate ontogenetic shifts in their diet where they consume larger prey as they grow older and their gape size increases. This shift enables squamates—especially snakes—to exploit new resources in their environments, usually mammalian prey. To test for ontogenetic variation in prey odor responses of a threatened snake species, the Northern pine snake (Pituophis melanoleucus melanoleucus), we presented food-naïve neonates and food-experienced adults with potential prey and non-prey animal scents and quantified their behavioral responses. Our data indicate a strong response to rodent scents from both neonates and adults. Further, neonates showed more frequent investigative probing and retreating behaviors from scented swabs and a higher rate of tongue-flicking than adults. We also developed a new metric for measuring snake responses to prey odor, a tongue-flick reaction score (TFRS), that incorporates investigative behaviors that may be unique to constrictor-type snakes. The TFRS did not differ between age classes and was highest when rodent odors were tested. A canonical discriminant analysis confirmed the relationship between TFRS behavioral components and tested chemical signal reactions. Based on our data, P. melanoleucus may fall into a category of snakes that exhibit an ontogenetic telescope rather than a general ontogenetic shift in their prey odor responses.

Airborne Chemical Information and Context-Dependent Post-Strike Foraging Behavior in Pacific Rattlesnakes (Crotalus oreganus)

Predators demonstrate context-dependent foraging behaviors to dynamically and successfully track prey and can use multiple cues in this process. In squamate reptiles (snakes and lizards), chemical signals from prey significantly influence predatory behavior, especially substrate and airborne cues. In this study, we examined behavioral variation in rattlesnakes (Crotalus oreganus) during strike-induced chemosensory searching (SICS), a sterotyped complex of behaviors seen in squamates. Rattlesnakes can use both substrate and airborne chemical cues during SICS, but we sought to determine the changes during SICS when either substrate, airborne, or air-deposited chemical cues were the only types available to snakes in a Y-maze. We hypothesize that these cues represent the spectrum of chemical information available in the natural environment. We also modified scoring of choice in the Y-maze by deriving a choice penalty score, a reflection of how extensively the snake explored the unscented arm of the maze. In the presence of substrate trails, rattlesnakes relocated prey fastest, had highest rates of tongue-flicking, and received the lowest choice penalty scores during SICS. Airborne chemical cues enabled successful relocation, but rattlesnakes took longer to relocate prey, increased the frequency of many searching behaviors, and more extensively explored the Y-maze (more negative choice penalties). When air-deposited cues were the only type available, rattlesnakes took the longest to choose an arm, had the lowest rates of tongue-flicking, and backtracked most often. We suggest that as prey odor becomes more dilute, rattlesnakes demonstrate behavioral plasticity in SICS to preserve their ability to relocate prey.