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Abstract Quantification of genital variation in males and females can inform our understanding of likely copulatory interactions and evolution of genital diversity. However, no studies have quantified genital shape variation within a single snake species or examined the shape and size of both the vaginal pouch and hemipenes. Here, we examine the shape and size of the genitalia of female and male diamondback water snakes, Nerodia rhombifer, using a three-dimensional automated landmark geometric morphometric approach on models of the lumen of the vaginal pouch and inflated hemipenes, applying these techniques for the first time to the genital shape of vertebrates. Vaginal pouch shape is significantly associated with body size and reproductive status. As females grow larger and become reproductive, the vaginal pouch enlarges, widens and becomes more bifurcated. In reproductive males, the shape of the hemipenes is also significantly associated with body size. As males grow larger, the hemipenes enlarge and widen; their bifurcation becomes more defined and the spines at the base become more prominent. Vaginal pouch and hemipenial centroid size are isometric with respect to body length. The centroid sizes of the hemipenes and vaginal pouch are not significantly different from one another, hence the genitalia match in size. Reproductive females and males covary in the degree of bifurcation and size of their genitalia. We demonstrate the utility of three-dimensional analysis in studies of the shape of soft tissues and advocate its use in future studies of genitalia.more » « less
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Abstract The gut microbial communities of mammals provide numerous benefits to their hosts. However, given the recent development of the microbiome field, we still lack a thorough understanding of the variety of ecological and evolutionary factors that structure these communities across species. Metabarcoding is a powerful technique that allows for multiple microbial ecology questions to be investigated simultaneously. Here, we employed DNA metabarcoding techniques, predictive metagenomics, and culture-dependent techniques to inventory the gut microbial communities of several species of rodent collected from the same environment that employ different natural feeding strategies [granivorous pocket mice (Chaetodipus penicillatus); granivorous kangaroo rats (Dipodomys merriami); herbivorous woodrats (Neotoma albigula); omnivorous cactus mice (Peromyscus eremicus); and insectivorous grasshopper mice (Onychomys torridus)]. Of particular interest were shifts in gut microbial communities in rodent species with herbivorous and insectivorous diets, given the high amounts of indigestible fibers and chitinous exoskeleton in these diets, respectively. We found that herbivorous woodrats harbored the greatest microbial diversity. Granivorous pocket mice and kangaroo rats had the highest abundances of the genus Ruminococcus and highest predicted abundances of genes related to the digestion of fiber, representing potential adaptations in these species to the fiber content of seeds and the limitations to digestion given their small body size. Insectivorous grasshopper mice exhibited the greatest inter-individual variation in the membership of their microbiomes, and also exhibited the highest predicted abundances of chitin-degrading genes. Culture-based approaches identified 178 microbial isolates (primarily Bacillus and Enterococcus), with some capable of degrading cellulose and chitin. We observed several instances of strain-level diversity in these metabolic capabilities across isolates, somewhat highlighting the limitations and hidden diversity underlying DNA metabarcoding techniques. However, these methods offer power in allowing the investigation of several questions concurrently, thus enhancing our understanding of gut microbial ecology.more » « less
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Abstract BackgroundSnakes exhibit extreme intestinal regeneration following months-long fasts that involves unparalleled increases in metabolism, function, and tissue growth, but the specific molecular control of this process is unknown. Understanding the mechanisms that coordinate these regenerative phenotypes provides valuable opportunities to understand critical pathways that may control vertebrate regeneration and novel perspectives on vertebrate regenerative capacities. ResultsHere, we integrate a comprehensive set of phenotypic, transcriptomic, proteomic, and phosphoproteomic data from boa constrictors to identify the mechanisms that orchestrate shifts in metabolism, nutrient uptake, and cellular stress to direct phases of the regenerative response. We identify specific temporal patterns of metabolic, stress response, and growth pathway activation that direct regeneration and provide evidence for multiple key central regulatory molecules kinases that integrate these signals, including major conserved pathways like mTOR signaling and the unfolded protein response. ConclusionCollectively, our results identify a novel switch-like role of stress responses in intestinal regeneration that forms a primary regulatory hub facilitating organ regeneration and could point to potential pathways to understand regenerative capacity in vertebrates.more » « less