Pheromonal communication in urodelan amphibians.

Pheromonal communication is an ancient and pervasive sensory modality in urodelan amphibians. One family of salamander pheromones (the sodefrin precursor-like factor (SPF) family) originated 300 million years ago, at the origin of amphibians. Although salamanders are often thought of as relatively simple animals especially when compared to mammals, the pheromonal systems are varied and complex with nuanced effects on behavior. Here, we review the function and evolution of pheromonal signals involved in male-female reproductive interactions. After describing common themes of salamander pheromonal communication, we describe what is known about the rich diversity of pheromonal communication in each salamander family. Several pheromones have been described, ranging from simple, invariant molecules to complex, variable blends of pheromones. While some pheromones elicit overt behavioral responses, others have more nuanced effects. Pheromonal signals have diversified within salamander lineages and have experienced rapid evolution. Once receptors have been matched to pheromonal ligands, rapid advance can be made to better understand the olfactory detection and processing of salamander pheromones. In particular, a large number of salamander species deliver pheromones across the skin of females, perhaps reflecting a novel mode of pheromonal communication. At the end of our review, we list some of the many intriguing unanswered questions. We hope that this review will inspire a new generation of scientists to pursue work in this rewarding field.

Presence of sodefrin precursor-like factor pheromone candidates in mental and dorsal tail base glands in the plethodontid salamander, Karsenia koreana.

Plethodontid salamanders are well known for their distinct courtship rituals and the associated pheromonal signaling. However, little is known about pheromones produced in the lone Asian plethodontid species Karsenia koreana. Here, we examined the localization patterns of proteins of the sodefrin precursor-like factor (SPF) pheromone system in K. koreana. Using an antibody generated against SPF proteins from another plethodontid, Desmognathus ocoee, we tested three types of skin glands in K. koreana males via immunohistochemistry: the mental gland and two types of dorsal tail base glands-caudal courtship glands and dorsal granular glands. SPF immunoreactivity was detected in the known courtship gland, the mental gland, as well as granular glands, but not in caudal courtship glands. Due to immunoreaction specificity, we hypothesize the proteins of the SPF system in K. koreana and D. ocoee are structurally and functionally related and are used as courtship pheromones in K. koreana. Also, we hypothesize that K. koreana males transmit SPF to the female during the tail-straddling walk via dorsal granular glands. Finally, K. koreana male caudal courtship glands may be producing SPF proteins that are not recognized by our SPF antibody or these glands may play a different role in courtship than anticipated.

The Evolution of Derived Monomorphism From Sexual Dimorphism: A Case Study on Salamanders.

While sexual dimorphism has long received special attention from biologists, derived monomorphism, the condition in which both males and females express similar derived features has been less well studied. Historically, the appearance of "male-like" features in females has been explained by the genetic correlation between the sexes. Recent work emphasizes the importance of studying the independent selective forces on both females and males to understand sexual dimorphism. Sexual dimorphism and derived monomorphism in the genus Aneides are examined in light of predictions of social selection. Aneides hardii shows the greatest degree of sexual dimorphism in snout-vent length and head width, with the other species of Aneides less sexually dimorphic. This reduced dimorphism, however, is not a return to an ancestral monomorphic state, but rather exemplifies derived monomorphism because females express traits that were limited in expression to males of ancestral species. Instead of calling these "male-typical" traits in females, I suggest the term "derived monomorphic" traits as these traits are typical in these females, and "derived monomorphic" can apply to both sexes. Increased attention to studying the patterns and ecological significance of derived monomorphism will shed light on the underlying selective forces, including sexual selection, on both females and males.

Spanish Resumen El estudio del dimorfismo sexual ha recibido amplia atención en la biología. En cambio, el monomorfismo derivado, la condición en la que tanto los machos como las hembras expresan características derivadas similares, ha sido menos estudiado. Históricamente, la aparición de características "masculinas" en las hembras se ha explicado como el resultado de la correlación genética entre los sexos. Avances recientes han enfatizado la importancia de estudiar las fuerzas selectivas en machos y hembras independientemente. Éste trabajo analiza el dimorfismo sexual y el monomorfismo derivado en el género Aneides considerando predicciones de selección social. La especie Aneides hardii tiene el mayor grado de dimorfismo sexual en cuanto a la longitud hocico-cloaca y ancho de cabeza, mientras que las otras especies de Aneides son menos dimórficas. Sin embargo, este dimorfismo reducido no es una regresión a un estado monomórfico ancestral, sino un monomorfismo derivado puesto que las hembras expresan rasgos que se expresaban exclusivamente en los machos en especies ancestrales. Dado que estos rasgos son típicos en éstas hembras, propongo que en lugar de llamar a éstos rasgos "típicos masculinos," se use el término de rasgos "monomórficos derivados", que puede usarse para ambos sexos. El estudio de los patrones e importancia ecológica del monomorfismo derivado permitirá entender las fuerzas selectivas subyacentes, incluyendo la selección sexual, tanto en hembras como en machos. Translated to Spanish by Eleanor JS Weisblat and Laura Diaz-Martinez.

A New Perspective on Female-to-Male Communication in Salamander Courtship.

Courtship behavior in salamanders is often complex and involves well-documented communication from males to females in multiple sensory modalities. Historically, behaviors exhibited during the major stages of courtship have been predominately framed as a male acting and signaling to "persuade" a passive female to participate in courtship and remain with him until sperm release is completed. In this review, we use courtship descriptions for lungless salamanders (Plethodontidae) as a case study to illustrate this historical bias of a male-centered perspective. We then re-examine the literature and summarize the many ways females are active participants during plethodontid courtships. We also relate female behaviors to the types of female-to-male communication that may occur. For example, females have been documented to approach a male and initiate courtship, participate in mutual head rubbing, and step astride the male's tail to begin the tail-straddling walk (a key courtship behavior observed in all plethodontids). Additionally, females have glands that may produce chemical signals that males respond to during courtship. We conclude that communication during courtship is more accurately described as a two-way interaction where each partner's behavior is coordinated with the other's via multi-modal signaling. Shifting the lens through which we view courtship and behavior provides insight into which female behaviors and anatomical features are most likely to be used for communication with males.

It Takes Two to Tango: Including a Female Perspective in Reproductive Biology.

Like many scientific disciplines, the field of reproductive biology is subject to biases in terminology and research foci. For example, females are often described as coy and passive players in reproductive behaviors and are termed "promiscuous" if they engage in extra-pair copulations. Males on the other hand are viewed as actively holding territories and fighting with other males. Males are termed "multiply mating" if they mate with multiple females. Similarly, textbooks often illustrate meiosis as it occurs in males but not females. This edition of Integrative and Comparative Biology (ICB) includes a series of papers that focus on reproduction from the female perspective. These papers represent a subset of the work presented in our symposium and complementary sessions on female reproductive biology. In this round table discussion, we use a question and answer format to leverage the diverse perspectives and voices involved with the symposium in an exploration of theoretical, cultural, pedagogical, and scientific issues related to the study of female biology. We hope this dialog will provide a stepping-stone toward moving reproductive science and teaching to a more inclusive and objective framework.

Multi-Institutional, Multidisciplinary Study of the Impact of Course-Based Research Experiences.

Numerous national reports have called for reforming laboratory courses so that all students experience the research process. In response, many course-based research experiences (CREs) have been developed and implemented. Research on the impact of these CREs suggests that student benefits can be similar to those of traditional apprentice-model research experiences. However, most assessments of CREs have been in individual courses at individual institutions or across institutions using the same CRE model. Furthermore, which structures and components of CREs result in the greatest student gains is unknown. We explored the impact of different CRE models in different contexts on student self-reported gains in understanding, skills, and professional development using the Classroom Undergraduate Research Experience (CURE) survey. Our analysis included 49 courses developed and taught at seven diverse institutions. Overall, students reported greater gains for all benefits when compared with the reported national means for the Survey of Undergraduate Research Experiences (SURE). Two aspects of these CREs were associated with greater student gains: 1) CREs that were the focus of the entire course or that more fully integrated modules within a traditional laboratory and 2) CREs that had a higher degree of student input and results that were unknown to both students and faculty.

Scaling Up: Adapting a Phage-Hunting Course to Increase Participation of First-Year Students in Research.

Authentic research experiences are valuable components of effective undergraduate education. Research experiences during the first years of college are especially critical to increase persistence in science, technology, engineering, and mathematics fields. The Science Education Alliance Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) model provides a high-impact research experience to first-year students but is usually available to a limited number of students, and its implementation is costly in faculty time and laboratory space. To offer a research experience to all students taking introductory biology at Gonzaga University (n = 350/yr), we modified the traditional two-semester SEA-PHAGES course by streamlining the first-semester Phage Discovery lab and integrating the second SEA-PHAGES semester into other courses in the biology curriculum. Because most students in the introductory course are not biology majors, the Phage Discovery semester may be their only encounter with research. To discover whether students benefit from the first semester alone, we assessed the effects of the one-semester Phage Discovery course on students' understanding of course content. Specifically, students showed improvement in knowledge of bacteriophages, lab math skills, and understanding experimental design and interpretation. They also reported learning gains and benefits comparable with other course-based research experiences. Responses to open-ended questions suggest that students experienced this course as a true undergraduate research experience.

In situ localization of plethodontid courtship pheromone mRNA in formalin-fixed tissue.

Male plethodontid salamanders produce courtship pheromones that increase female receptivity. Three protein components of the courtship pheromone cocktail have been characterized in the mental gland of Plethodon shermani, the red-legged salamander: plethodontid receptivity factor (PRF), plethodontid modulating factor (PMF), and sodefrin precursor-like factor (SPF). In this study, a streamlined in situ hybridization (ISH) protocol, employing a biotinylated oligonucleotide probe, is used to visualize the sites of pheromone expression in formalin-fixed paraffin-embedded P. shermani mental gland and post-cloacal tail tissue. Results corroborate previous RT-PCR studies on pheromone expression. PRF and PMF are highly expressed in P. shermani mental gland, while SPF expression is more variable. None of the tested pheromones is expressed in dorsal or ventral tail glands. The reported protocol is simple, rapid, and effective, allowing visualization of high-copy mRNA transcript in formalin-fixed tissue.