Among vertebrate gastrointestinal microbiome studies, complete representation of taxa is limited, particularly among reptiles. Here, we provide evidence for previously unrecognized host-microbiome associations along the gastrointestinal tract from the American alligator, a crown archosaur with shared ancestry to extinct taxa, including dinosaurs. Microbiome compositional variations reveal that the digestive system consists of multiple, longitudinally heterogeneous microbiomes that strongly correlate to specific gastrointestinal tract organs, regardless of rearing histories or feeding status. A core alligator gut microbiome comprised of Fusobacteria, but depleted in Bacteroidetes and Proteobacteria common to mammalians, is compositionally unique from other vertebrate gut microbiomes, including other reptiles, fish, and herbivorous and carnivorous mammals. As such, modern alligator gut microbiomes advance our understanding of archosaur gut microbiome evolution, particularly if conserved host ecology has retained archosaur-specific symbioses over geologic time.

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Reptiles and fish have robust regenerative powers for tooth renewal. However, extant mammals can either renew their teeth one time (diphyodont dentition) or not at all (monophyodont dentition). Humans replace their milk teeth with permanent teeth and then lose their ability for tooth renewal. Here, we study tooth renewal in a crocodilian model, the American alligator, which has well-organized teeth similar to mammals but can still undergo life-long renewal. Each alligator tooth is a complex family unit composed of the functional tooth, successional tooth, and dental lamina. Using multiple mitotic labeling, we map putative stem cells to the distal enlarged bulge of the dental lamina that contains quiescent odontogenic progenitors that can be activated during physiological exfoliation or artificial extraction. Tooth cycle initiation correlates with β-catenin activation and soluble frizzled-related protein 1 disappearance in the bulge. The dermal niche adjacent to the dermal lamina dynamically expresses neural cell adhesion molecule, tenascin-C, and other molecules. Furthermore, in development, asymmetric β-catenin localization leads to the formation of a heterochronous and complex tooth family unit configuration. Understanding how these signaling molecules interact in tooth development in this model may help us to learn how to stimulate growth of adult teeth in mammals.Wnt | placode | slow cycling | regeneration

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Eggs were sampled from 22 wild American alligator nests from the Rockefeller Wildlife Refuge in south-west Louisiana, along with the females guarding the nests. Three nests were sampled in 1995 and 19 were sampled in 1997. Females and offspring from all clutches were genotyped using five polymorphic microsatellite loci and the three nests from 1995 were also genotyped using one allozyme locus. Genotypes of the hatchlings were consistent with the guarding females being the mothers of their respective clutches. Multiple paternity was found in seven of the 22 clutches with one being fathered by three males, and the remaining six clutches having genotypes consistent with two males per clutch. Paternal contributions of multiply sired clutches were skewed. Some males sired hatchlings of more than one of the 22 clutches either as one of two sires of a multiple paternity clutch, as the sole sire of two different clutches, or as the sole sire of one clutch and one of two sires of a multiply sired clutch. There was no significant difference between females that had multiple paternity clutches and those that had singly sired clutches with respect to female total length (P = 0.844) and clutch size (P = 0.861). Also, there was no significant correlation between genetic relatedness of nesting females and pairwise nest distances (r2 = 0.003, F1,208 = 0.623, P = 0.431), indicating that females in this sample that nested close to one another were no more related than any two nesting females chosen at random. Eleven mutations were detected among hatchlings at the five loci over the 22 clutches. Most of these mutations (eight of 11) occurred at Ami(mu)-17, the only compound microsatellite locus of the five used in this study, corresponding to a mutation rate of 1.7 x 10-3. Finally, most of the mutations (82%) were homoplasious, i.e., mutating to an allelic state already present in this Louisiana population.

The concept that reptiles regulate their body temperature by behavioural means, such as shuttling between sun and shade (Cowles and Bogert, 1944;Hertz et al., 1993), has become widely accepted in vertebrate thermal physiology. Behavioural adjustments enable many diurnal species of reptile to maintain high and stable body temperatures in the face of fluctuations in environmental temperatures (Avery, 1982;Seebacher et al., 1999). The importance of body temperature regulation is seen to lie in maximising the rates of temperature-sensitive physiological functions (Huey, 1982). The rates of chemical reactions, including those catalysed by enzymes, are dependent on the energetic state of the compounds involved, which in turn is strongly influenced by temperature. The rates of most physiological processes are, therefore, a direct function of the temperature of the organism. Thermoregulation that includes high metabolic heat production combined with effective insulation often allows endotherms to maintain an elevated body temperature within a narrow range (Lovegrove et al., 1991). The low metabolic rates of reptiles make metabolic heat production negligible, and regulation of body temperature is achieved by behavioural means such as microhabitat selection (Cowles Reptiles living in heterogeneous thermal environments are often thought to show behavioural thermoregulation or to become inactive when environmental conditions prevent the achievement of preferred body temperatures. By contrast, thermally homogeneous environments preclude behavioural thermoregulation, and ectotherms inhabiting these environments (particularly fish in which branchial respiration requires body temperature to follow water temperature) modify their biochemical capacities in response to long-term seasonal temperature fluctuations. Reptiles may also be active at seasonally varying body temperatures and could, therefore, gain selective advantages from regulating biochemical capacities. Hence, we tested the hypothesis that a reptile (the American alligator Alligator mississippiensis) that experiences pronounced seasonal fluctuations in body temperature will show seasonal acclimatisation in the activity of its metabolic enzymes. We measured body temperatures of alligators in the wild in winter and summer (N=7 alligators in each season), and we collected muscle samples from wild alligators (N=31 in each season) for analysis of metabolic enzyme activity (lactate dehydrogenase, citrate synthase and cytochrome c oxidase). There were significant differences in mean daily body temperatures between winter (15.66±0.43°C; mean ± S.E.M.) and summer (29.34±0.21°C), and daily body temperatures fluctuated significantly more in winter compared with summer. Alligators compensated for lower winter temperatures by increasing enzyme activities, and the activities of cytochrome c oxidase and lactate dehydrogenase were significantly greater in winter compared with summer at all assay temperatures. The activity of citrate synthase was significantly greater in the winter s...

The normalisation of average muscle architecture data for adult American alligators (Alligator mississippiensis of +10 Kg body mass) was performed using an incorrect divisor. Rather than normalisation of muscle architecture data using division by derivations of body mass (body mass 0.67 for area measures such as muscle physiological cross-sectional area [PCSA], and body mass 0.33 for linear measurements such as muscle fascicle lengths), data from an older dataset normalised using division by derivations of body volume were mistakenly included in the final manuscript.In Table 2 (pp. 427-428) and

Mammals and birds have a specialized cardiac atrioventricular conduction system enabling rapid activation of both ventricles. This system may have evolved together with high heart rates to support their endothermic state (warm-bloodedness) and is seemingly lacking in ectothermic vertebrates from which first mammals then birds independently evolved. Here, we studied the conduction system in crocodiles (Alligator mississippiensis), the only ectothermic vertebrates with a full ventricular septum. We identified homologues of mammalian conduction system markers (Tbx3-Tbx5, Scn5a, Gja5, Nppa-Nppb) and show the presence of a functional atrioventricular bundle. The ventricular Purkinje network, however, was absent and slow ventricular conduction relied on trabecular myocardium, as it does in other ectothermic vertebrates. We propose the evolution of the atrioventricular bundle followed full ventricular septum formation prior to the development of high heart rates and endothermy. In contrast, the evolution of the ventricular Purkinje network is strongly associated with high heart rates and endothermy.