Interactions between Endothelin Receptor B and Transcription Factors Sox10 and Pax3 in the Melanocyte Lineage

Genetic interactions that underlie developmental processes such as cell differentiation and pattern formation are complex and difficult to elucidate. Neural Crest (NC) cells and their derivatives offer an optimal system in which to probe for these complex interactions as they acquire different cell fates and constitute a variety of structures. The transcription factors Sox10 and Pax3 as well as the transmembrane receptor Endothelin receptor b (Ednrb) are temporally and spatially co-expressed early in NC cells and mutations in these genes lead to similar hypopigmentation phenotypes due to a reduced number of NC-derived melanocyte precursors, the melanoblasts. The goal of this study was to establish whether Sox10 and Ednrb or Pax3 and Ednrb interact to promote normal murine melanocyte development. Crosses of Sox10 or Pax3 with Ednrb heterozygous mutants showed that the double heterozygous hypopigmentation phenotype was significantly more pronounced than phenotypes of single heterozygotes, implying that a synergistic interaction exists between Sox10 and Ednrb and Pax3 and Ednrb. This interaction was further explored by the attempt to rescue the Sox10 and Pax3 hypopigmentation phenotypes by the transgenic addition of Ednrb to melanoblasts. Pigmentation was completely restored in the Sox10 and partially restored in the Pax3 mutant mice. The comparison of the number of melanoblasts in transgenic and non-transgenic Sox10 mutant embryos showed that the transgenic rescue occurred as early as E11.5, a critical time for melanoblast population expansion. Cell survival assays indicated that the rescue was not due to an effect of the transgene on melanoblast survival. A novel phenotype arose when studying the interaction between Ednrb and Pax3. Newborns appeared normal but by 3.5 weeks of age, the affected pups were smaller than normal littermates and developed a dome-shaped head; some also developed thoracic kyphosis. Affected pups were dead by 4 weeks of age: 80% were Pax3Sp/+ and 75% were female. When compared to normal littermates, affected mice had brains with enlarged 4th ventricles and more glia while skeletal staining showed kyphosis, wider rib cages and pelvic differences. An epistatic interaction resulting from the mixing of genetic backgrounds that is exacerbated in the presence of Pax3 heterozygosity is suspected.

Ultrastructural Basis and Developmental Control of Blue Iridescence in Selaginella Leaves

The iridescentb lue color of several Selaginellasp ecies is caused by a physical effect, thinfilm interference.P redictionsf or a model film have been confirmedb y electronm icroscopyo f S. willdenowaEnid S. uncinataF. or the latters pecies iridescencec ontributest o leaf absorption at wavelengths above 450 nm and develops in environments enriched with far-red (730 nm) light. This evidence supports the involvement of phytochrome in the developmental control of iridescence.

The Maintenance and Consequences of a Low Quality Diet in Poecilia Latipinna

The adaptive significance of herbivory in nature is not well understood. In order to document the conditions that select for an herbivorous feeding habit, we must first understand how such a diet is maintained, and the consequences of doing so. A few studies have begun to reveal mechanisms of maintaining herbivory (i.e. selective feeding, diet mixing, etc.) and the associated life history responses (i.e. growth, reproduction, etc.) in terrestrial and marine systems; however, studies of this kind are underrepresented in the freshwater literature. In this study, I use the sailfin molly (Poecilia latipinna) as a model organism to examine diet selectivity and the effects of an herbivorous diet on growth. To study food selectivity, sailfin mollies were fed either disturbed or intact periphyton mats from one of three localities within the Everglades (Water Conservation Area 3B, the Gap, or Chekika). Mats are structured with palatable algal species (i.e. greens and diatoms) comprising the inner components of the mat, and unpalatable species (i.e. cyanobacteria) comprising the outer edges. Fish gut contents were analyzed for each treatment and periphyton locality. Results suggest that when provided access to the inner components of the mats, fish preferentially eat more palatable algae. In a second experiment, effects of an herbivorous diet were examined using neonate sailfin mollies. Fish were fed either commercial food flakes, commercial algae flakes, or ground periphyton, and growth rate was measured weekly, from birth to 21 days. Fish fed the commercial diets grew at a faster rate and reached a larger final size than those fed periphyton. These results suggest that a periphyton diet is limited in nutritional elements compared to a pure algae diet and herbivorous organisms feeding upon it may experience negative effects on growth. By studying the costs and benefits of herbivory in a freshwater system, this paper contributes to a larger study of the question of why herbivory would evolve as an adaptation when seemingly inefficient compared to carnivorous and omnivorous diets.

Otras mitologías (Other Mythologies) and Catch and Release , Book Presentation by Author Reina María Rodríguez

This flyer promotes the event "Otras mitologías (Other Mythologies) and Catch and Release : Book Presentation by Author Reina María Rodríguez" cosponsored by the School of International and Public Affairs and the Department of Modern Languages and Florida International University. The event was held and Books & Books in Coral Gables.

Density-Dependent Effects of Coral Reef Restoration on Herbivory and Community Interactions

Coral reefs are among the most productive ecosystems in the world. Yet, with their recent declines due to disease, climate change, and overfishing, restoration of these habitats is one of the main concerns for ecologists, resource managers, and government organizations. Coral reef restoration aims to promote key ecosystem processes to shift these habitats to their historical state of high coral cover, but few studies have focused on effective ways to promote resilience. In addition, little is known about the impact of restoration on the fish communities. The aim of this study is to understand how the community of herbivorous fishes is affected by the density of coral outplants inside a special protection area located in the Florida Keys National Marine Sanctuary. Grazing rates, number of visits and time spent foraging were compared using video footage of sites previously devoid of corals, and six months after coral restorations had occurred. Coral transplantations did not appear to attract herbivores nor increase grazing rates of fishes. Instead Sparisoma and Acanthurus fishes appear to respond to changes in the environment by modifying their grazing behavior. However, there was an observed increase in visits by Acanthurus species after transplantation for all the sites sampled within the reef. These fishes seemed to prefer low coral cover sites for grazing. This study highlights the importance of examining coral restorations impacts at the community level. Understanding how restoration influences herbivores and other guilds of reef fishes will allow individuals to not only determine if these habitats are returning to their “original” state, but provide more information on the ways these systems cope with changes in the environment.

A Comparative Study of Concurrent Acoustic and Diver Survey Data, and Fish Community Descriptions of a High Latitude Coral Reef, Florida, USA

Fisheries independent data on relatively unstudied nekton communities were used to explore the efficacy of new tools to be applied in the investigation of shallow coastal coral reef habitats. These data obtained through concurrent diver visual and acoustic surveys provided descriptions of spatial community distribution patterns across seasonal temporal scales in a previously undocumented region. Fish density estimates by both diver and acoustic methodologies showed a general agreement in ability to detect distributional patterns across reef tracts, though magnitude of density estimates were different. Fish communities in southeastern Florida showed significant trends in spatial distribution and seasonal abundance, with higher estimates of biomass obtained in the dry season. Further, community composition shifted across reef tracts and seasons as a function of the movements of several key reef species.

Non-Consumptive Effects of Predators in Coral Reef Communities and the Indirect Consequences of Marine Protected Areas

Predators exert strong direct and indirect effects on ecological communities by intimidating their prey. Non-consumptive effects (NCEs) of predators are important features of many ecosystems and have changed the way we understand predator-prey interactions, but are not well understood in some systems. For my dissertation research I combined a variety of approaches to examine the effect of predation risk on herbivore foraging and reproductive behaviors in a coral reef ecosystem. In the first part of my dissertation, I investigated how diet and territoriality of herbivorous fish varied across multiple reefs with different levels of predator biomass in the Florida Keys National Marine Sanctuary. I show that both predator and damselfish abundance impacted diet diversity within populations for two herbivores in different ways. Additionally, reef protection and the associated recovery of large predators appeared to shape the trade-off reef herbivores made between territory size and quality. In the second part of my dissertation, I investigated context-dependent causal linkages between predation risk, herbivore foraging behavior and resource consumption in multiple field experiments. I found that reef complexity, predator hunting mode, light availability and prey hunger influenced prey perception of threat and their willingness to feed. This research argues for more emphasis on the role of predation risk in affecting individual herbivore foraging behavior in order to understand the implications of human-mediated predator removal and recovery in coral reef ecosystems.

Effects of Multiple Ecological Drivers on Recruitment and Succession of Coral Reef Macroalgal Communities

The study evaluated the effects of herbivory pressure, nutrient availability and potential propagule supply on recruitment and succession of coral reef macroalgal communities. Recruitment and succession tiles were placed in a nutrient-herbivory factorial experiment and macroalgal abundances were evaluated through time. Proportional abundances of macroalgal form-functional groups on recruitment and succession tiles were similar to field established communities within treatments, evidencing possible effects of adult macroalgae as propagule supply. Macroalgal abundance of recruitment tiles increased with nutrient loading and herbivory reduction combined whereas on succession tiles nutrient loading increased abundance of articulated-calcareous only when herbivores were excluded. Macroalgal field established communities were only affected by herbivory reduction.

Using stable isotopes to investigate trophic interactions of bottlenose dolphins (Tursiops truncatus) in the Florida Coastal Everglades

Top predators are known for their ability to 1) affect their communities through predation and 2) induce behavioral modifications. Recent research suggests that they may also play “bottom-up” roles in ecosystems, including transporting materials within and across habitat boundaries. The Florida Coastal Everglades (FCE) is an “upside-down” oligotrophic estuary where productivity decreases from the mouth of the estuary to freshwater marshes. Therefore, movements of predators may be important in ecosystem dynamics. While other estuarine predators in the FCE have been shown to potentially move nutrients among ecosystems, the potential for bottlenose dolphins (Tursiops truncatus) to play a similar role in the systems has not been investigated. Stable isotope analysis of biopsy samples were used to investigate spatial variation in trophic interactions of dolphins to see if they might transport nutrients. Values of δ15 N suggest dolphins feed at a trophic level similar to other top predators in the ecosystem while δ13 C suggest that dolphins forage largely within food webs where they were sampled rather than transporting nutrients across ecosystem boundaries. The exception may be dolphins foraging in rivers, which may transport nutrients downstream; a pattern opposite to that of bull sharks and alligators in these habitats. Further research is necessary to predict how future changes occurring due to restoration and climate will affect the ecological roles of dolphins.

Bioinformatic analysis for exploring relationships between genes and gene products

To carry out their specific roles in the cell, genes and gene products often work together in groups, forming many relationships among themselves and with other molecules. Such relationships include physical protein-protein interaction relationships, regulatory relationships, metabolic relationships, genetic relationships, and much more. With advances in science and technology, some high throughput technologies have been developed to simultaneously detect tens of thousands of pairwise protein-protein interactions and protein-DNA interactions. However, the data generated by high throughput methods are prone to noise. Furthermore, the technology itself has its limitations, and cannot detect all kinds of relationships between genes and their products. Thus there is a pressing need to investigate all kinds of relationships and their roles in a living system using bioinformatic approaches, and is a central challenge in Computational Biology and Systems Biology. This dissertation focuses on exploring relationships between genes and gene products using bioinformatic approaches. Specifically, we consider problems related to regulatory relationships, protein-protein interactions, and semantic relationships between genes. A regulatory element is an important pattern or "signal", often located in the promoter of a gene, which is used in the process of turning a gene "on" or "off". Predicting regulatory elements is a key step in exploring the regulatory relationships between genes and gene products. In this dissertation, we consider the problem of improving the prediction of regulatory elements by using comparative genomics data. With regard to protein-protein interactions, we have developed bioinformatics techniques to estimate support for the data on these interactions. While protein-protein interactions and regulatory relationships can be detected by high throughput biological techniques, there is another type of relationship called semantic relationship that cannot be detected by a single technique, but can be inferred using multiple sources of biological data. The contributions of this thesis involved the development and application of a set of bioinformatic approaches that address the challenges mentioned above. These included (i) an EM-based algorithm that improves the prediction of regulatory elements using comparative genomics data, (ii) an approach for estimating the support of protein-protein interaction data, with application to functional annotation of genes, (iii) a novel method for inferring functional network of genes, and (iv) techniques for clustering genes using multi-source data.

Multiparameter flow cytometric analysis of C -reactive protein binding to human-derived cell lines and peripheral blood monocytes

C-reactive protein (CRP), a normally occurring human plasma protein may become elevated as much as 1,000 fold during disease states involving acute inflammation or tissue damage. Through its binding to phosphorylcholine in the presence of calcium, CRP has been shown to potentiate the activation of complement, stimulate phagocytosis and opsonize certain microorganisms. Utilizing a flow cytometric functional ligand binding assay I have demonstrated that a monocyte population in human peripheral blood and specific human-derived myelomonocytic cell lines reproducibly bind an evolutionarily conserved conformational pentraxin epitope on human CRP through a mechanism that does not involve its ligand, phosphorylcholine. ^ A variety of cell lines at different stages of differentiation were examined. The monocytic cell line, THP-1, bound the most CRP followed by U937 and KG-1a cells. The HL-60 cell line was induced towards either the granulocyte or monocyte pathway with DMSO or PMA, respectively. Untreated HL-60 cells or DMSO-treated cells did not bind CRP while cells treated with PMA showed increased binding of CRP, similar to U-937 cells. T cell and B-cell derived lines were negative. ^ Inhibition studies with Limulin and human SAP demonstrated that the binding site is a conserved pentraxin epitope. The calcium requirement necessary for binding to occur indicated that the cells recognize a conformational form of CRP. Phosphorylcholine did not inhibit the reaction therefore the possibility that CRP had bound to damaged membranes with exposed PC sites was discounted. ^ A study of 81 normal donors using flow cytometry demonstrated that a majority of peripheral blood monocytes (67.9 ± 1.3, mean ± sem) bound CRP. The percentage of binding was normally distributed and not affected by gender, age or ethnicity. Whole blood obtained from donors representing a variety of disease states showed a significant reduction in the level of CRP bound by monocytes in those donors classified with infection, inflammation or cancer. This reduction in monocyte populations binding CRP did not correlate with the concentration of plasma CRP. ^ The ability of monocytes to specifically bind CRP combined with the binding reactivity of the protein itself to a variety of phosphorylcholine containing substances may represent an important bridge between innate and adaptive immunity. ^