No capítulo 1 os principais fatores envolvidos com a distribuição geográfica do gênero Enyalius foram a temperatura média anual, a latitude e a amplitude média da temperatura diurna, indicando uma possível adaptação termal do gênero. A distribuição potencial do grupo refletiu na variação geográfica na morfologia, como demonstrado pelas diferenças no tamanho do corpo e estruturas morfológicas, que foram explicadas por variáveis climáticas, espaciais e estruturais do ambiente. A regra de Allen corroborou com os padrões da variação na morfologia de Enyalius, que foram correlacionados com as condições térmicas e geográficas. Os padrões de variação morfológica também foram associados com cobertura vegetal (e talvez a disponibilidade de água nas diferentes regiões), que pode exercer influência sobre a adaptação morfológica de Enyalius e explicar parte do crescimento diferencial alométrico sobre a vasta área geográfica. A hipótese de adaptação térmica deve ser a base para explicar a distribuição de Enyalius em ambientes de florestas do Brasil. No capítulo 2 estudamos as áreas de distribruição potencial de cada espécie do grupo e encontramos para a maioria delas características ambientais gerais que foram importantes para a ocorrência do gênero nos seus ambientes. A latitude e a amplitude média da temperatura diurna, que se sobreporam na maioria dos modelos de nicho potencial, ao longo da costa brasileira, deve caracterizar a distribuição principal das espécies de Enyalius no Domínio Florestal Atlântico. Isso nos indicou um “padrão geral” de fatores básicos importantes para a ocorrência do grupo neste bioma e que, provavelmente, parte dos nichos ecológicos similares entre algumas espécies, em geral, ocorreram por influência ambiental de ampla magnitude em associação à plasticidade fenotípica das espécies como resposta ecológica adaptativa. No capítulo 3, nós encontramos algumas respostas para as espécies de Enyalius, havendo padrões morfológicos e ambientais específicos associados independentemente da relação filogenética ou da proximidade geográfica em que estas se encontram. Acreditamos que as características ambientais e o uso destas, como recursos, devem influenciar mais sobre a ocorrência das espécies em seus hábitats (pressão de seleção natural) do que por uma questão de inércia filogenética herdada ao grupo. Este estudo também indicou que determinadas mudanças evolutivas nas proporções e no tamanho do corpo das espécies de Enyalius devem estar associadas à divergência de hábitats e suas características e/ou ao uso diferente de substratos entre as espécies de lagartos, e que, consequentemente, necessitam de maiores investigações para o conhecimento das causas das relações ecológicas reveladas aqui. Por fim, os padrões observados sobre a variação morfológica nas espécies de Enyalius devem ser uma conseqüência direta de respostas adaptativas a ação das condições ambientais locais (e.g. seleção natural), o que por sua vez nos revelou grande plasticidade adaptativa das espécies através de suas ecologias em ambientes de continente. / In the first chapter, we saw that the main factors affecting the Enyalius’ geographical distribution were the annual mean temperature, latitude, and the mean diurnal temperature, which indicates a possible thermal adaptation as a base to explain the group distribution. Enyalius distribution was reflected in the morphological geographical variation, as demonstrated by the differences in the body size and morphological structures, which were explained by climatic, spatial and structural variables. The Allen rule was corroborated, as patterns in the morphological variation in Enyalius were correlated with thermal and geographical conditions. The thermal adaptation hypothesis may be the basis to explain Enyalius distribution in different Brazilian forests. In the second chapter we studied the potential distribution areas for each species in the group, and we identified the general environmental characteristics affecting the species occurrence in their habitats, for most species. We suggest that variables such as latitude and mean amplitude of daily temperature along the Brazilian coast overlapped in most of our models, characterizing the main distribution of the species belonging to the genus Enyalius in the Atlantic Forest. In the third chapter, we found that there are morphological and environmental patterns associated with the species, independent of their phylogenetic or geographical relationship. We observed that environmental features seem to be more critical to determine the species occurrence than their evolutionary heritage (e.g. philogenetic inertia). Finally, this study indicated that some evolutionary changes in body size and proportions might be associated to the divergence in habitat requirements and/or the differences in substrate use, which need further investigation. We conclude that the observed morphological variation in Enyalius can be a direct consequence of adaptative responses to local environmental conditions, which demonstrated the high phenotipic plasticity, based on their ecology in environments from mainland.

abstract: Well-established model systems exist in four out of the seven major classes of vertebrates. These include the mouse, chicken, frog and zebrafish. Noticeably missing from this list is a reptilian model organism for comparative studies between the vertebrates and for studies of biological processes unique to reptiles. To help fill in this gap the green anole lizard, Anolis carolinensis, is being adapted as a model organism. Despite the recent release of the complete genomic sequence of the A. carolinensis, the lizard lacks some resources to aid researchers in their studies. Particularly, the lack of transcriptomic resources for lizard has made it difficult to identify genes complete with alternative splice forms and untranslated regions (UTRs). As part of this work the genome annotation for A. carolinensis was improved through next generation sequencing and assembly of the transcriptomes from 14 different adult and embryonic tissues. This revised annotation of the lizard will improve comparative studies between vertebrates, as well as studies within A. carolinensis itself, by providing more accurate gene models, which provide the bases for molecular studies. To demonstrate the utility of the improved annotations and reptilian model organism, the developmental process of somitogenesis in the lizard was analyzed and compared with other vertebrates. This study identified several key features both divergent and convergent between the vertebrates, which was not previously known before analysis of a reptilian model organism. The improved genome annotations have also allowed for molecular studies of tail regeneration in the lizard. With the annotation of 3' UTR sequences and next generation sequencing, it is now possible to do expressional studies of miRNA and predict their mRNA target transcripts at genomic scale. Through next generation small RNA sequencing and subsequent analysis, several differentially expressed miRNAs were identified in the regenerating tail, suggesting miRNA may play a key role in regulating this process in lizards. Through miRNA target prediction several key biological pathways were identified as potentially under the regulation of miRNAs during tail regeneration. In total, this work has both helped advance A. carolinensis as model system and displayed the utility of a reptilian model system. / Dissertation/Thesis / Ph.D. Molecular and Cellular Biology 2012

No capítulo 1 os principais fatores envolvidos com a distribuição geográfica do gênero Enyalius foram a temperatura média anual, a latitude e a amplitude média da temperatura diurna, indicando uma possível adaptação termal do gênero. A distribuição potencial do grupo refletiu na variação geográfica na morfologia, como demonstrado pelas diferenças no tamanho do corpo e estruturas morfológicas, que foram explicadas por variáveis climáticas, espaciais e estruturais do ambiente. A regra de Allen corroborou com os padrões da variação na morfologia de Enyalius, que foram correlacionados com as condições térmicas e geográficas. Os padrões de variação morfológica também foram associados com cobertura vegetal (e talvez a disponibilidade de água nas diferentes regiões), que pode exercer influência sobre a adaptação morfológica de Enyalius e explicar parte do crescimento diferencial alométrico sobre a vasta área geográfica. A hipótese de adaptação térmica deve ser a base para explicar a distribuição de Enyalius em ambientes de florestas do Brasil. No capítulo 2 estudamos as áreas de distribruição potencial de cada espécie do grupo e encontramos para a maioria delas características ambientais gerais que foram importantes para a ocorrência do gênero nos seus ambientes. A latitude e a amplitude média da temperatura diurna, que se sobreporam na maioria dos modelos de nicho potencial, ao longo da costa brasileira, deve caracterizar a distribuição principal das espécies de Enyalius no Domínio Florestal Atlântico. Isso nos indicou um “padrão geral” de fatores básicos importantes para a ocorrência do grupo neste bioma e que, provavelmente, parte dos nichos ecológicos similares entre algumas espécies, em geral, ocorreram por influência ambiental de ampla magnitude em associação à plasticidade fenotípica das espécies como resposta ecológica adaptativa. No capítulo 3, nós encontramos algumas respostas para as espécies de Enyalius, havendo padrões morfológicos e ambientais específicos associados independentemente da relação filogenética ou da proximidade geográfica em que estas se encontram. Acreditamos que as características ambientais e o uso destas, como recursos, devem influenciar mais sobre a ocorrência das espécies em seus hábitats (pressão de seleção natural) do que por uma questão de inércia filogenética herdada ao grupo. Este estudo também indicou que determinadas mudanças evolutivas nas proporções e no tamanho do corpo das espécies de Enyalius devem estar associadas à divergência de hábitats e suas características e/ou ao uso diferente de substratos entre as espécies de lagartos, e que, consequentemente, necessitam de maiores investigações para o conhecimento das causas das relações ecológicas reveladas aqui. Por fim, os padrões observados sobre a variação morfológica nas espécies de Enyalius devem ser uma conseqüência direta de respostas adaptativas a ação das condições ambientais locais (e.g. seleção natural), o que por sua vez nos revelou grande plasticidade adaptativa das espécies através de suas ecologias em ambientes de continente. / In the first chapter, we saw that the main factors affecting the Enyalius’ geographical distribution were the annual mean temperature, latitude, and the mean diurnal temperature, which indicates a possible thermal adaptation as a base to explain the group distribution. Enyalius distribution was reflected in the morphological geographical variation, as demonstrated by the differences in the body size and morphological structures, which were explained by climatic, spatial and structural variables. The Allen rule was corroborated, as patterns in the morphological variation in Enyalius were correlated with thermal and geographical conditions. The thermal adaptation hypothesis may be the basis to explain Enyalius distribution in different Brazilian forests. In the second chapter we studied the potential distribution areas for each species in the group, and we identified the general environmental characteristics affecting the species occurrence in their habitats, for most species. We suggest that variables such as latitude and mean amplitude of daily temperature along the Brazilian coast overlapped in most of our models, characterizing the main distribution of the species belonging to the genus Enyalius in the Atlantic Forest. In the third chapter, we found that there are morphological and environmental patterns associated with the species, independent of their phylogenetic or geographical relationship. We observed that environmental features seem to be more critical to determine the species occurrence than their evolutionary heritage (e.g. philogenetic inertia). Finally, this study indicated that some evolutionary changes in body size and proportions might be associated to the divergence in habitat requirements and/or the differences in substrate use, which need further investigation. We conclude that the observed morphological variation in Enyalius can be a direct consequence of adaptative responses to local environmental conditions, which demonstrated the high phenotipic plasticity, based on their ecology in environments from mainland.

L'anthropisation croissante et les changements globaux entrainent de nombreuses perturbations dans l'environnement conduisant à la dégradation, et même à la destruction d'habitat. Le modèle allostatique proposé par McEwen et Wingfield en 2003 permet de mieux appréhender les relations entre physiologie et réponses démographique dans le cadre de la physiologie de la conservation. A travers des approches expérimentales en milieux controlé et semi-naturel nous avons testé l'effet de d'un stress chronique sur les changements des traits d'histoires de vie et les paramétres physiologique chez le lézard vivipare (Zootoca vivipara). Nous discutons de la caractérisation de la charge allostatique et des effets à court terme et long terme d'un stress chronique et l'utilisation de la charge allostatique comme indicateur de la dégradation de l'habitat. Ce travail pourra être utilisé afin de mieux comprendre et prédire la dynamique des populations naturelles soumises aux changements globaux. / Increasing anthropogenic and global changes are causing many disturbances in the environment leading to degradation and even destruction of habitat. The allostatic model proposed by McEwen and Wingfield in 2003 provides a better understanding of the relationships between physiology and demographic responses within the framework of conservation physiology. Experimental approaches in controlled and semi-natural environments have tested the effect of chronic stress on changes in life histories and physiological parameters in the viviparous lizard (Zootoca vivipara). We discuss the characterization of allostatic charge and the short-term and long-term effects of chronic stress and the use of allostatic charge as an indicator of habitat degradation. This work can be used to better understand and predict the dynamics of natural populations subjected to global changes.

abstract: Traumatic injury to the central nervous or musculoskeletal system in traditional amniote models, such as mouse and chicken, is permanent with long-term physiological and functional effects. However, among amniotes, the ability to regrow complex, multi-tissue structures is unique to non-avian reptiles. Structural regeneration is extensively studied in lizards, with most species able to regrow a functional tail. The lizard regenerated tail includes the spinal cord, cartilage, de novo muscle, vasculature, and skin, and unlike mammals, these tissues can be replaced in lizards as adults. These studies focus on the events that occur before and after the tail regrowth phase, identifying conserved mechanisms that enable functional tail regeneration in the green anole lizard, Anolis carolinensis. An examination of coordinated interactions between peripheral nerves, Schwann cells, and skeletal muscle reveal that reformation of the lizard neuromuscular system is dependent upon developmental programs as well as those unique to the adult during late stages of regeneration. On the other hand, transcriptomic analysis of the early injury response identified many immunoregulatory genes that may be essential for inhibiting fibrosis and initiating regenerative programs. Lastly, an anatomical and histological study of regrown alligator tails reveal that regenerative capacity varies between different reptile groups, providing comparative opportunities within amniotes and across vertebrates. In order to identify mechanisms that limit regeneration, these cross-species analyses will be critical. Taken together, these studies serve as a foundation for future experimental work that will reveal the interplay between reparative and regenerative mechanisms in adult amniotes with translational implications for medical therapies. / Dissertation/Thesis / Differentially Expressed Genes in the Early Regenerating Lizard Tail / Gene Ontology of Differentially Expressed Genes in the Early Regenerating Lizard Tail / KEGG and Reactome Pathway Analysis of Differentially Expressed Genes in the Early Regenerating Lizard Tail / 3D Reconstruction of an Alligator Regenerated Endoskeleton / Lateral 2D Serial Sections of a Regenerated Alligator Tail / Doctoral Dissertation Biology 2020

No description available.

abstract: Satellite cells are adult muscle stem cells that activate, proliferate, and differentiate into myofibers upon muscle damage. Satellite cells can be cultured and manipulated in vitro, and thus represent an accessible model for studying skeletal muscle biology, and a potential source of autologous stem cells for regenerative medicine. This work summarizes efforts to further understanding of satellite cell biology, using novel model organisms, bioengineering, and molecular and cellular approaches. Lizards are evolutionarily the closest vertebrates to humans that regenerate entire appendages. An analysis of lizard myoprogenitor cell transcriptome determined they were most transcriptionally similar to mammalian satellite cells. Further examination showed that among genes with the highest level of expression in lizard satellite cells were an increased number of regulators of chondrogenesis. In micromass culture, lizard satellite cells formed nodules that expressed chondrogenic regulatory genes, thus demonstrating increased musculoskeletal plasticity. However, to exploit satellite cells for therapeutics, development of an ex vivo culture is necessary. This work investigates whether substrates composed of extracellular matrix (ECM) proteins, as either coatings or hydrogels, can support expansion of this population whilst maintaining their myogenic potency. Stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, while the ECM composition was not significantly important. Additionally, satellite cells on hydrogels entered a quiescent state that could be reversed when the cells were subsequently cultured on Matrigel. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for satellite cells. To further understand how different signaling pathways control satellite cell behavior, an investigation of the Notch inhibitor protein Numb was carried out. Numb deficient satellite cells fail to activate, proliferate and participate in muscle repair. Examination of Numb isoform expression in satellite cells and embryonic tissues revealed that while developing limb bud, neural tube, and heart express the long and short isoforms of NUMB, satellite cells predominantly express the short isoforms. A preliminary immunoprecipitation- proteomics experiment suggested that the roles of NUMB in satellite cells are related to cell cycle modulation, cytoskeleton dynamics, and regulation of transcription factors necessary for satellite cell function. / Dissertation/Thesis / Doctoral Dissertation Molecular and Cellular Biology 2020

Historically, Australia has been important in the study of, and the development of hypotheses aimed at understanding, the evolution of viviparity in amniote vertebrates. Part of the importance of Australia in the field results from a rich fauna of skinks, including one of the broadest ranges of diversity of placental structures within one geographic region. During the last decade, we have focussed our studies on one lineage, the Eugongylus group of skinks of the subfamily Lygosominae because it contains oviparous species and some that exhibit complex placentae. Our specific objective has been to attempt to understand the fundamental steps required when viviparity, and ultimately complex placentae, evolve from oviparous ancestors. We have taken a three-prong approach: (1) detailed study of the morphology and ontogeny of the placentae of key species at the light microscope level; (2) study of changes in the uterus associated with pregnancy, or the plasma membrane transformation; and (3) measures of the net exchange of nutrients across the placenta or eggshell of key species. In turn, we have found that: (1) details of the morphology and ontogeny of placentae are more complex that originally envisaged, and that the early conclusions about a sequence in the evolution of complex placentae was naïve; (2) a plasma membrane transformation occurs in viviparous, but not oviparous lizards, and thus may be a fundamental feature of the evolution of viviparity in amniotes; and (3) species with more complex chorioallantoic placentae tend to transport more nutrients across the placenta during pregnancy than those with simpler chorioallantoic placentae but, because the correlation is not tight, the importance of the omphaloplacenta in transporting nutrients may have been overlooked. Also, the composition of yolk of highly matrotrophic species is broadly similar, but not identical, to the yolk of oviparous species. Some of the interpretation of our data within the context of our specific objective is not yet possible, pending the publication of a robust phylogeny of Eugongylus group skinks. Once such a phylogeny is available, we are in a position to propose specific hypotheses about the evolution of viviparity that can be tested using another lineage of amniotes, possibly Mabuya group skinks. © 2002 Elsevier Science Inc. All rights reserved.

The San Luis Valley is a large valley formation in Colorado surrounded on either side by mountain ranges exceeding 4,267 m. Within the Valley, two of the 14 amphibian and reptile species are dwarfed: the short-horned lizard (Phrynosoma hernandesi) and the Great Plains toad (Anaxyrus cognatus). Since its initial reporting in 1968 and confirmation in 1981, no research further investigating this dwarfism has been conducted. I collected morphological measurements to determine the extent and patterns of dwarfism of both species. I then investigated the genetics of both species using mitochondrial DNA to determine whether they are genetically distinct, their colonization histories within the Valley, and whether the Valley functions as a reproductive barrier. Lastly, I report life/natural-history data to determine the effects of dwarfism. Phrynosoma hernandesi and A. cognatus were significantly dwarfed and showed an increase in sexual size dimorphism compared to populations surrounding the Valley. Valley populations of P. hernandesi show high amounts of genetic divergence from populations surrounding the Valley while A. cognatus shows minimal genetic variation throughout its range. Based on the variable distribution of genetic variation in the Valley, historic climate patterns, and fossil records, there are two most likely colonization histories for P. hernandesi: 1.) populations colonized the Valley during a singular event and have since diverged or 2.) populations colonized the Valley during two events that correlate with the two warm, dry periods within the last 0.8 MYA. Dwarfed P. hernandesi consumed diets similar to populations outside the Valley although there is local variation in the diversity of prey items consumed. Phrynosoma hernandesi at Zapata Ranch showed annual variation in body size and morphology while population dynamics correlate with the timing of precipitation. Also, females show a reduced reproductive output, producing fewer neonates but of equal size to non-dwarfed neonates. Collectively, findings from this study suggest that Valley populations represent unique taxa and should be considered for further genetic study to determine their taxonomic and conservation status.

The effect of total metabolic cost expenditures on the precision of behavioral thermoregulation was investigated for the purportedly eurythermic Northern Alligator lizard (Gerrhonotus coeruleus). An operant apparatus was designed to test metabolic output at different heat reinforcement magnitudes. The mean TB reflected in each trial was positively correlated to the length of reinforcement. The shuttle rate during each trial was inversely correlated to the length of reinforcement. The standard deviation and total metabolic costs did not vary significantly between trials undertaken at the same ambient temperature. Eurythermality in G. coeruleus is caused by fluctuations in preferred body temperature and not by fluctuations around this temperature. The metabolic cost of behavioral thermoregulation did not change with corresponding changes in reinforcement magnitude. This indicates that eurythermality is the recorded effect of lizards behaviorally regulating to different TB in a stenothermic manner, rather than of random TB fluctuations in a wide range of normal activity (the classical view of eurythermality).